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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
The books in the series are designed as easy-to-use guides to the microorganisms found in foods. Alison Turner. for his scientific input and Victoria Emerton. I thank the contributing authors for their collaboration. thank you for all the support. Fish and Seafood. guides to HACCP. potential hazards associated with the consumption of a range of products. Rhea Fernandes Leatherhead Food International v . and Meat Products. All handbooks also contain a review of the related legislation in Europe and UK.FOREWORD The Microbiology Handbook series includes Dairy Products. My gratitude is extended to Dr Peter Wareing. published by Leatherhead Food International and RSC Publishing. Training Manager. which has been the chief factor in ensuring the completion of this book.Goldwyn. and Ann Pernet for the work put into typesetting and indexing. Technical Team Leader for her valuable editorial contribution. Each book provides a brief overview of the processing factors that determine the nature and extent of microbial growth and survival in the product. I also acknowledge and thank Jackie Apps. and a detailed list of contacts for various food authorities. I dedicate this book to my husband . both in the UK and elsewhere. Acknowledgements All the authors involved in writing the first edition of this book are gratefully acknowledged. 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.
1 Introduction 2. vii .1 Definitions 3.3 Initial Microflora 2.5 Pathogens: Growth and Survival 3.6 Pathogens: Growth and Survival 2.5 Pathogens: Growth and Survival 4. 4.4 Spoilage 4.3 Processing and its Effects on the Microflora 3.CONTENTS CONTRIBUTORS FOREWORD INTRODUCTION 1.8 Further Reading iii v xi 1 1 3 5 9 12 15 19 20 24 27 27 28 31 32 43 45 47 49 53 53 59 60 65 66 70 71 77 79 79 80 81 83 84 87 89 92 2.8 Further Reading CRUSTACEAN SHELLFISH 4.4 Spoilage 3.8 References MOLLUSCAN SHELLFISH 3.3 Processing and its Effects on the Microflora 4.9 Further Reading CHILLED AND FROZEN PREPARED FISH PRODUCTS 2.1 Definitions 1.5 Factors Affecting Fresh Fish Spoilage 1.7 References 4.2 Initial Microflora 4.6 Published Microbiological Criteria 4.7 Published Microbiological Criteria 2.2 Definitions 2.6 Pathogens: Growth and Survival 1.1 Definitions 4.6 Published Microbiological Criteria 3.2 Initial Microflora 3.8 References 1. CHILLED AND FROZEN RAW FISH 1.4 Processing and its Effects on the Microflora 2.5 Spoilage 2.7 References 3. 3.2 Initial Microflora 1.4 Spoilage 1.7 Published Microbiological Criteria 1.3 Processing and its Effects on the Microflora 1.
2 Legislative Structure 190 9. 7.4 Spoilage 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. 853/2004 Laying Down Specific Hygiene Rules for Food of Animal Origin 196 9.2 Initial Microflora 6.1 Definitions 5.4 Spoilage 5. CURED.2 Definitions 8. SMOKED AND DRIED FISH 5.9 Other Relevant Legislation 223 9.10 Cyclic Imines 7. 852/2004 on the General Hygiene of Foodstuffs 191 9.5.4 Regulation (EC) No.5 Pathogens: Growth and Survival 6.1 Introduction 7. 2073/2005 on Microbiological Criteria for Foodstuffs 214 9.7 References 5. 2006 No.4 Amnesic Shellfish Poisoning (ASP) Toxins 7.2 Paralytic Shellfish Poisoning (PSP) Toxins 7.6 Published Microbiological Criteria 5.12 References HACCP IN FISH AND SEAFOOD PRODUCT MANUFACTURE 8.3 Tetrodotoxin 7.7 Neurotoxin Shellfish Poisoning (NSP) Toxins 7.5 Pathogens: Growth and Survival 5. S. EU FOOD HYGIENE LEGISLATION 189 9.7 Food Hygiene (England) Regulations 2006.2 Initial Microflora 5.5 Diarrhetic Shellfish Poisoning (DSP) Toxins 7.I.8 Ciguatoxins 7.1 Introduction 8.6 References 6.4 Implementation and Review of the HACCP Plan 8.3 Stages of a HACCP Study 8.7 Further Reading FISH AND SHELLFISH TOXINS 7.3 Processing and its Effects on the Microflora 5.5 References 93 93 95 96 108 113 116 117 120 123 123 126 127 135 136 137 140 141 141 142 144 146 148 151 153 155 158 160 165 166 175 175 176 177 184 186 6.6 Regulation (EC) No.10 Further Reading 223 viii .3 Regulation (EC) No. 14 (Hygiene requirements specific to the UK) 221 9.6 Lipophilic Shellfish Toxins (LST) 7. 8. 9.8 Further Reading FERMENTED FISH 6.11 Conclusion 7.1 Definitions 6.1 Introduction 189 9.9 Azaspiracids 7.3 Processing and its Effects on the Microflora 6.8 Guidance 222 9.5 Regulation (EC) No.
3 Clostridium perfringens 10.11 References 225 225 226 227 229 230 230 232 233 234 235 236 241 247 CONTACTS INDEX ix . 10.6 Salmonella spp.10 Vibrio vulnificus 10. 10.10 PATHOGEN PROFILES 10.1 Aeromonas spp.2 Clostridium botulinum 10.8 Vibrio cholerae 10.7 Staphylococcus aureus 10. 10.9 Vibrio parahaemolyticus 10.4 Listeria spp.5 Plesiomonas 10.
Poultry. fish and seafood can be involved in the transmission of pathogenic microorganisms and toxins.Fish and Seafood consists of the microbiology of seven different product categories: chilled and frozen raw fish. in Handbook of Food Products Manufacturing: Health. The Microbiology Handbook. 2006. Seafood processing: basic sanitation practices. the consumption of fish and seafood has increased. 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. Wageningen. fermented fish. 94767. crustacean shellfish. Catches can be gathered from seas. The second edition of this handbook is a review of the entire book for currency of information. season. thus. Chandan R.B.INTRODUCTION Fish and seafood are a main source of animal protein in the diet. Eds.. Key changes in this edition are the recent regulatory changes pertaining to food hygiene and microbiological criteria for foodstuffs. and. Geographical region.M. chilled and frozen prepared fish. Nollet L.A.. Hui Y.. Often contamination is from human and animal sources. Luten J. Cross N. Hoboken. Seafood and Vegetables. smoked and dried fish. Because of their health advantages over red meats. Jacobsen C. Oehlenschlager J. molluscan shellfish. Milk. for fish. Clark S.. Seafood Research from Fish to Dish: Quality. Safety and Processing of Wild and Farmed Fish.. rivers and lakes whose water can range from pristine to contaminated.H. Bekaert K. Saebo A.. cured. and fish and shellfish toxins.. Meat.. Wageningen Academic Publishers. xi . 2007. Further Reading General Fish/Seafood Microbiology Cross N. Wiley.
Hui Y. Kluwer Academic / Plenum Publishers. Salmonella spp. Hafsteinsson H.. in Food Processing: Principles and Applications.Y. 543-61. Spoilage and shelf-life extension of fresh fish and shellfish. Huss H.International Commission on Microbiological Specifications for Foods. 2005. 1997. Johnson J. 431-43. 1997.A. and Clostridium botulinum in smoked fish and shellfish. Incidence and survival of Listeria monocytogenes in ready-to-eat seafood products. Heinitz M. Gram L.H. Gram L. Taylor S.. Oxford. Gorham J. 87121.. Flick G.H.B. 1998. Fish and fish products (Microorganisms in fish products). in Fish Processing Technology.J. Ed. Woodhead Publishing Ltd.M.L.E.P. in Advances in Food and Nutrition Research. Ashie I. Blackwell Publishing. Biogenic amines in fish and shellfish. Safety and Quality Issues in Fish Processing. 174-249. in Seafood from Producer to Consumer: Integrated Approach to Quality. 1997.L. Stanfield P. November 1995. Modified-atmosphere packaging of fish and fish products.. Food Control. Woodhead Publishing Ltd. Vo1ume 39. Microbiological spoilage of fish and fish products. Elsevier... Blech Z. Amsterdam. in Food Plant Sanitation. Marcel Dekker. Ohlsson T. International Commission on Microbiological Specifications for Foods. 1997.. 329-65. Eds. Bengtsson N. CRC Critical Reviews in Food Science and Nutrition.H. Control of indigenous pathogenic bacteria in seafood. 36 (1-2). 245-66 Bremner H. Tong P. 8 (2).K.A.. Blackwell Publishing. November 1995.. Luten J.. Gudmundsson M. Ventresca P. Hui Y. Cambridge. Blech Z. Ed. Stanfield P. Academic Press. Ed. Luten J. Rawles D. Eds. Minimal processing in practice: seafood. Noordwijkerhout. 372-6. 2002.. Smith J. Amsterdam. Oxford. Hall G.Y. 1996. London..B. Ed. 91-8. Blackie. Proceedings of the International Seafood Conference. Huss H.S. 121-37. Eds. Noordwijkerhout. in Kosher Food Production.D. 60 (4). basic sanitation practices. Nip W-K. xii . Proceedings of the International Seafood Conference. 167-77. 1996. 61 (3). in Microorganisms in Foods 6: Microbial Ecology of Food Commodities.R.. Bruinsma B. Huss H. Martin R. Simpson B. Davies A. Microbiology of fish and fish products.A. Seafood processing: basic sanitation practices. Journal of Food Protection. 2002.R. 2004. 200-23. 1997. 413-30. 459-72. Ed. Predictive microbiological modelling and seafood quality.L.S.H. International Journal of Food Microbiology. The fish industry. Elsevier. in Minimal Processing Technologies in the Food Industry. London..M. Dalgaard P.. Journal of Food Protection.. 2003. Seafood: processing. Smith J. New York. Ed. 318-23. Cambridge. New York.N. 2004. in Seafood from Producer to Consumer: Integrated Approach to Quality. 33 (1). Dalgaard P.. The incidence of Listeria spp.H.. 1996. McCarthy S.
. 1996. 166-73.D. London. Dutson T. Liston J. Sofos J. Jakobsen M. 118-20.R. Olley J. 305-17. Bonnell A. in Public Health Aspects of Seafood-borne Zoonotic Diseases. 1991. August 1991. 46 (1). Glasgow.. 26 (3). Twigg B. Copenhagen. 1995. Blackie. PHLS Microbiology Digest. 1990. 319-33. Fish spoilage and safety. 25-39. Eds.R.. Garrett E. Ross T. Jakobsen M. The microbiology of seafood products.M. Gram L.M. and specifications for seafood products. Microbiological indicators and seafood processing. 1-17. poultry and fish. Eds. 5 (3). Activities Report of the R&D Associates. 269-82. Liston J.S. Elsevier. 26 (3). 1990. 54-61. guidelines. Ed. International Journal of Food Microbiology. 1994. Hobbs G.R. 5662. Application of predictive microbiology to assure the quality and safety of fish and fish products. in Quality Assurance in the Fish Industry: Proceedings of an International Conference. Evaluation of the bacteriological quality of seafood. Ed. Microbial growth and its control in meat. Poultry and Fish Products. Huss H.H. 1994. Amsterdam. 1992. Bernoth E-M. 93-105.A.N. Elsevier. xiii . International Journal of Food Microbiology.D. Bonnell A. Evaluation of the bacteriological quality of seafood. 2257. 90-106. Dalgaard P. Chapman and Hall. Amsterdam. Ward D. 1994. 265-79. International Journal of Food Microbiology. A new look at microbiological standards. in Quality Assurance in the Fish Industry: Proceedings of an International Conference. 1992. Chapman & Hall.M. Ward D. Blackie. 121-8. Ed. Liston J. Ed. in Quality Attributes and Their Measurement in Meat. Gram L. London. Ed. Fish: microbiological spoilage and safety. Martin A. Huss H. Sterling Publications Ltd. Eds. London.H. August 1991. Dalgaard P. Huss H. in Microorganisms in Foods. Copenhagen. 1989.International Commission on Microbiological Specifications for Foods.. 1994.A. in Quality Assurance in the Fish Industry: Proceedings of an International Conference.. Pearson A. 359-403. August 1991. Food Science and Technology Today. 1994. Bacterial spoilage of seafood. Microbiological quality of fishery products... Qualitative and quantitative characterization of spoilage bacteria from packed fish. London. Amsterdam. 16 (1). Institut fur Veterinarmedizin. International Commission on Microbiological Specifications for Foods. McMeekin T.J. Microbial hazards of seafood consumption. in Food and Beverage Technology International: USA.. 1995. Elsevier. Gibson D. Liston J. 44 (12). Gilbert R. Liston J. 459-78. Berlin. 1992.H. Jakobsen M. Bacterial pathogens transmitted by seafood. Food Technology. Copenhagen.. Seafood toxins of microbiological origin. 1992. Eds. 11 (2). Modelling of microbial activity and prediction of shelf-life for packed fish. in Quality Assurance in Seafood Processing: A Practical Guide. Volume 5: Microbiological Specifications of Food Pathogens. in Fisheries Processing: Biotechnological Applications.
Eyles M. Seafood-borne bacterial pathogens of marine origin. Chichester. Shelf-life extension of fresh fish .Pedrosa-Menabrito A.R. 1982. 71-117. Regenstein J. Hobbs G. 1988. Hackney C. 11 (2). CSIRO Food Research Quarterly.S. Journal of Food Quality. 1986. Food Technology. Microbiology of frozen fish and related products.K. 104-9. 42 (3).spoilage of fish.R. guidelines. 1987. Robinson R. Hobbs G. The bacteriology of fish handling and processing. 8-16. Ed. Ed. Wiley.C.a review. Ed. Hodgkiss W. Lamprecht E. in Developments in Food Microbiology. London. 1988. xiv . 117-27. Norris J. Volume 1.K. Elsevier Applied Science Publishers. Microbiological standards. Part I . Simmonds C. Garrett E. Dicharry A.M.. 1988. and specifications and inspection of seafood products. Davies R. 46 (1).. 169-208. 42 (3). London. Food Technology. 90-3. Applied Science Publishers.. 103. Microbiology of fish. in Essays in Agricultural and Food Microbiology. Microbiological hazards associated with fishery products.. in Microbiology of Frozen Foods. 1985.J. 199-226.
crustaceans.1 Definitions Fish are classified as any of the cold-blooded aquatic vertebrates of the super class Pisces typically showing gills. some regulatory agencies such as the US Food and Drug Administration (FDA) will include molluscan shellfish. fins and a streamline body. giving it its characteristically flaky structure. Fish fillet composition can vary significantly within the same species due to feed intake. lean fish are those fish that store most of their fat in the liver.21%. This super class of vertebrates includes all the bony and cartilaginous finfish. ‘fish’ also refers to the flesh of such animals used as food. and other forms of aquatic animal as part of their ‘fish’ definition. CHILLED AND FROZEN RAW FISH Associate Prof. Protein content in fish fillet varies typically from 16 . Fish are an important part of a healthy diet since they contain high quality protein. it shows a typical season pattern especially in migratory species such as herring or mackerel.2 . and spawning season. migratory patterns. which can be up to 67%. Although fish is broadly similar in composition and structure to meat there are a number of distinctive features. while fatty fish have fat cells distributed along their bodies. “fish” will be used for fresh and seawater finfish. The connective tissue content of fish is also lower than that found in meat. In this chapter. typically 3 and 15% of total weight. typically fluctuates between 0.5% (1). respectively (1). The lipid fraction is the component showing the greatest variation. Muscle composition and structure of fish also differ from those found in other meat. most fish contain omega 3-fatty acids and other essential nutrients. In addition. Covadonga Arias Department of Fisheries and Allied Aquacultures Auburn University Auburn Alabama 36849 United States of America 1. In addition.1.20%. 1 . but typically present a low fat percent when compared to other meats. offering less than 0. However. The lipid content. and is mostly interspersed between the muscle fibres. Fish flesh is dominated by the abundance of white muscle in relatively short segments. Fish can be divided into fatty and lean fish. and excludes molluscs and Crustacea. Fish fillets are a poor source of carbohydrates.
nitrogen (N2) and oxygen (O2). colour. transportation and sale.FISH AND SEAFOOD Chilled fish is fish that has been cooled to. Evisceration is the removal of the viscera from a fish. and maintained at or below 7 °C. Quality refers to palatability and organoleptic characteristics such as tenderness. during storage. and texture of the fish. spoilage microflora describes an association of microorganisms that. Fresh fish is raw fish that has not been processed. Raw fish refers to fish that has not been cooked but excludes fish treated with curing salts and/or subjected to fermentation. Frozen fish is fish that has been cooled to. marbling. and maintained at or below -2 °C (normally below -12 °C) during storage. transportation and sale. juiciness. transportation and sale. but no further control is exercised during storage. Spoilage describes changes that render fish objectionable to consumers. hence. and flavour based on the maturity. frozen or preserved. colour. firmness. Superchilled fish is fish that has been cooled to. 2 . Modified-atmosphere packaging (MAP) refers to packaging systems in which the natural gaseous environment around the product is intentionally replaced by other gases. This technique is primarily used for bulk storage. through their development on fish. Shelf life is defined as the time of storage before microbial spoilage of a fish is evident. at -2 to -4 °C. DMA is dimethyl amine. usually carbon dioxide (CO2). Controlled-atmosphere packaging (CAP) refers to packaging in an atmosphere where the composition of the gases is continuously controlled during storage. Spoilage potential is a measure of the propensity of microorganisms to render fish objectionable to consumers through the production of offensive metabolic byproducts. The proportion of each component is fixed when the mixture is introduced. renders that fish objectionable to consumers. Organoleptic refers to qualities such as appearance. odour and texture. but not below 3 °C during storage. and maintained at temperatures just below the freezing point.
3 . Moraxella. Gastrointestinal tracts and gills typically yield high bacteria numbers.2 Initial Microflora The subsurface flesh of live. It is crucial to mimic the environmental physicochemical parameters when isolating bacteria from fish. Clostridium. Fish harvested from clean and cold waters will present lower bacterial numbers than fish from eutrophic and/or warm waters. It is difficult to estimate how many microorganisms are typically associated with fish. and those that although not autochthonous to the aquatic environment.) are also common aquatic bacteria. molecular-based methods have not yet been used to address this issue. as with other vertebrates. posing a concern for seafoodborne illnesses (2). However. Human pathogenic bacteria can be part of the initial microflora of fish. some species (most Vibrios) require sodium chloride for growth. the natural fish microflora tends to reflect the microbial communities of the surrounding waters. although these are influenced by water quality and feed. Shewanella and Pseudomonas. whenever possible. On the contrary. since they heavily depend on the type of sample analysed and the protocol used for isolation. and typical of the fish flora. In general.I). are present there as result of contamination (anthropomorphic origin or other) or are introduced to the fish during harvest. Flavobacterium. microorganisms colonise the skin. In fact. More accurate. potential human pathogens may be present in both scenarios. Gram-positive organisms such as Bacillus.CHILLED AND FROZEN RAW FISH Vacuum packaging (VP) refers to packaging systems in which the air is evacuated and the package sealed. These pathogens can be divided into two groups: organisms naturally present on fish (Table 1. the season and the method of harvest.II) (3). Lactobacillus and coryneforms can also be found in varying proportions (1). For example. 1. The autochthonous bacterial flora of fish is dominated by Gram-negative genera including: Acinetobacter. healthy fish is considered sterile and should not present any bacteria or other microorganisms. Members of the families Vibrionaceae (Vibrio and Photobacterium) and the Aeromonadaceae (Aeromonas spp. gills and the gastrointestinal tract of fish. 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. processing or storage (Table 1. Micrococcus. standard culture-dependent methods can only recover between 1 to 10% of total bacteria present in any given sample. several culture media containing sodium chloride and more than one incubation temperature should be used.
Toxin levels 0. stream water.FISH AND SEAFOOD TABLE 1. and mud Ubiquitous. vulnificus) 5 . birds.106 cells/g. Intestinal track of terrestrial vertebrates Human origin Fecal contamination Ubiquitous in environment Shigella spp.0.1 µg toxin lethal dose Clostridium botulinum non-proteolytic type E Pathogenic Vibrio spp.26 °C 10 . during storage indigenous spoilage bacteria tend to outgrow potential pathogenic bacteria.106 cells/g (exception: V.37 °C Unknown Unknown TABLE 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 . 4 .102 101 .35 °C 8 .II Non-indigenous bacterial pathogens frequently present in fish (3) Organism Listeria monocytogenes* Primary habitat Soil.103 High (107 .I Indigenous bacterial pathogens typically present in fish (3) Organism Temperature range Estimated Minimum Infective Dose 00. However.>106 Salmonella spp. In addition. 4).1 .14 . human origin Minimum Infective Dose Variable depending on the strain (>102 cells/g) Staphylococcus aureus 105 . and risk associated with the consumption of seafood is low (2.19 µg/kg bodyweight from < 102 .37 °C High for most species.109 cells/g) * Some sources considered L. Vibrio cholerae Vibrio parahaemolyticus Vibrio vulnificus other vibrios Aeromonas spp. Plesiomonas shigelloides 3 . sewage. Escherichia coli Yersinia enterocolitica 101 . numbers of pathogenic bacteria in raw fish tend to be low. estuarine environments.
or traps. mackerel or herring. Used refrigerated brines can be contaminated with high numbers of psychrotrophic spoilage bacteria. with fish such as tuna and cod being more commonly eviscerated than sardines. If evisceration is carried out at sea. and their re-use will increase the crosscontamination of other fish with such microorganisms. digestion and off-flavours in uneviscerated fish. freezing facilities (-18 °C) may be used to prevent the catch from deteriorating. and the killing process is designed to minimise stress. Physical damage such as loss of scales.1 Processing and its Effects on the Microflora Capture. ropes. cortisone levels increase during prolonged stress and can alter the fillet quality. with very little control over the condition of the fish at the time of death or the duration of the killing process. Inadequate circulation of chilled brines may result in localised anaerobic growth of some microorganisms. and during harvest. decks. however. Increasingly. in recent decades. the fish may be stored in the vessel for periods ranging from just a few hours to several weeks in melting ice.CHILLED AND FROZEN RAW FISH Shelf life depends on the initial microflora on the fish. and spoilage. with the production of off-odours. 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. boxes and/or baskets.3 1. 1. The length of time that set nets have been in the water or the time trawlers’ nets are towed. icing or freezing. Of particular concern is the use of wooden or soiled plastic containers for storage and unloading at the quayside. During capture and storage. offering better control of fish health prior to. In eviscerated fish.a practice that may have both advantages and disadvantages. However. potential contaminants added during handling and processing. in which the bacterial load 5 . In addition. the cuttings provide areas of exposed flesh that are open to microbial attack. aquaculture practices have been expanding worldwide. The decision to eviscerate the catch at sea will depend greatly on the size of the fish and the duration of storage at sea. hook and line. birds and soil. and conditions of storage. chilled brine or refrigerated seawater at -2 °C. Fish may be eviscerated prior to storage at sea . The action of intestinal enzymes and activity of the gut bacteria on the flesh around the belly cavity may produce discolouration. and especially when fish is stored on board for longer periods. This contrasts greatly with the meat industry. bruising and bursting of the gut will increase the number of sites open for bacterial attack and spread. human hands and clothing. has an effect on the amount of stress and physical damage that the fish will suffer during capture. These contacts will not only increase the bacterial cross-contamination between fish batches but will introduce microorganisms from other sources such as humans. handling and processing Wild finfish are usually caught by net. After capture.3. in which the health of each animal can be assessed prior to slaughter.
). What is more. cleaning of the work environment. 7). Brochothrix thermosphacta and psychrotrophic lactic acid bacteria (LAB) can produce spoilage characteristics. As with all foods. often in the absence of adequate refrigeration. Monitoring of the seawater for algal growth in order to limit the risk of algal toxin ingestion. Anaerobic atmospheres have less effect on fresh fish shelf life. and minimising handling will all reduce microbial cross-contamination. 1. and specific spoilage organisms may use other terminal electron acceptors naturally present in the fish (trimethylamine-N-oxide (TMAO). These changes can be deleterious.3. but if applied properly should extend the life of the product. will result in fully anoxic conditions. Bacterial composition under MAP shifts from mostly Gram-negative to predominantly Gram-positive (lactic) bacteria. by actively altering the composition of the 6 . Both carbon dioxide and low pH inhibit the growth of the typical spoilage bacteria such as Pseudomonas and Shewanella. but isolate the food in an environment different from the bulk atmosphere. from water activity (aw) through to physical damage. in time. allowing more growth of spoilage organisms. Carbon dioxide is the most important gas in MAP of fish because of its bacteriostatic and fungistatic properties. In MAP. fish have a higher post mortem pH. oxygen etc. if there is some oxygen demand from the product. Low oxygen levels have been shown to substantially prolong the freshness and quality life of refrigerated seafood products. In the absence of oxygen. Physical barriers not only protect from physical damage. not part of the normal flora of the meat animals (8). and of the quality of the water used for ice and to wash fish. These containers are also used for displaying the fish during auction at the quayside. partial fermentation of sugars occur leading to lower pH. they are usually process contaminants. for example. and to limit the loss of quality (5). careful and sanitary handling during processing is required to reduce the risk of contamination with potential human pathogens. By vacuum packing foods. 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. However. nitrogen.2 Modified-atmosphere packaging A natural atmosphere rich in oxygen (21%) is responsible for oxidative processes and for all aerobic respiratory life. The atmospheric conditions surrounding a product may be passively or actively altered. the natural atmosphere is replaced with a controlled gas mixture (carbon dioxide. however. potential spoilage bacteria are among the psychrophilic and psychrotrophic flora present on temperate-water fish before death (9). ferric ion (Fe3+)).FISH AND SEAFOOD can be substantial. a reduction in the oxygen tension is achieved which. Packaging changes the intrinsic and extrinsic parameters affecting a product. use of effective detergents and disinfectants.
The principal effect of raised carbon dioxide-MAP is an extension of the 'lag' phase of the growth of the bacteria on the fish. the National Advisory Committee on Microbiological Criteria for Foods (NACMCF) has published a number of recommendations on 7 . without overt signs of spoilage. the higher pH and reduction potential (Eh) of the fish muscle. The products. nitrous oxide and carbon monoxide have also been suggested as possible replacement gases in trace amounts for MAP/CAP. Growth and toxin production have been detected in artificially contaminated packs of whole trout after 1 week’s incubation at 10 °C (13). although less information on their effectiveness is available. Of particular concern are the psychrotrophic type E and non-proteolytic type B and F strains. but only in recent years has it seen a marked expansion in use and market share. slower-growing flora (11).CHILLED AND FROZEN RAW FISH surrounding gas. including salt. a modified-atmosphere may contain any gas necessary for the desired effect. botulinum. and the MAP of fish has more challenges to overcome as a result of: a comparatively large initial load of bacteria present. In the United States. and increase in-pack drip (12). Micrococcus and Moraxella). Sulphur dioxide. which provides guidance relating to vacuum or modified-atmosphere packaging for specific fish products (14). The single most important concern with respect to the use of MAP is the potential for outgrowth and toxin production by C. but can dissolve into fish liquids and deform packages. The (International) Codex Committee has published a Code of Practice for Fish and Fishery Products (CAC/RCP 52-2003). MAP has been applied to fresh meat and fish with a resulting commercially viable extension in shelf life (10). after additive application. phosphates. and the problems of muscle structure damage by the modified-atmosphere (11). and the promotion of a predominantly Gram-positive. but residual oxygen and other electron acceptors may be sufficient to allow oxidative spoilage of fish (12). High concentrations of carbon dioxide have the most pronounced microbial effect. discolour pigmented fish (11). does inhibit some aerobic bacteria and reduce the rate of oxidative rancidity.3 °C and produce toxins. The microflora of meat is not the same as that of whole. Modified-atmosphere preservation of fish was first reported in the 1930s. VP is one of the oldest forms of altering the interior gaseous environment of a pack. an inert and odourless gas. may not be considered ‘fresh’ fish. Replacement of oxygen with nitrogen. gutted or filleted fish. Many additives have been tried in conjunction with changed atmosphere. sorbates and chelating agents such as ethylenediaminetetraacetic acid (EDTA) (11). This has been driven partly by increased consumer demand for fresh and chilled convenience foods containing fewer chemical preservatives. Flavobacterium. which are able to grow rapidly at low temperature. as they are able to grow at temperatures as low as 3. Gases used in MAP of fish most commonly include carbon dioxide and nitrogen. the inhibition of common 'spoilage' bacteria (Pseudomonas. the pathogens that may be able to grow before spoilage occurs.
3 °C at all points from packaging onwards. however. grouping of fish types is possible to estimate the best carbon dioxide level to be used in MAP. It highlights temperature control as the primary preventive measure against the possible hazard of toxin production by C. oxidative rancidity is a significant source of spoilage and hence the complete removal of oxygen and the use of oxygen-impermeable films are preferred. and the use of a HACCP plan. greater advantages are seen. this problem occurs to a lesser extent with meat. The addition of high levels of carbon dioxide to these fish. 3. The only processing dip to be used is 5% potassium sorbate and 10% sodium tripolyphosphate. Freshwater fish have different intrinsic bacterial flora and also do not contain such high concentrations of amines (e. from where drip loss and gaping of the muscle meant that organoleptically classified rejection occurred sooner than with no carbon dioxide addition (12). trimethylamine and sulfides). owing to the dissolution of cellular structures. should be used. Although all fish species need to be treated separately. The growth of these bacteria is reduced by high carbon dioxide concentrations but. Subsequent storage must be at or below 3 °C. leading to the recommendation that the sale of MAP/VP raw fishery products be allowed only when certain conditions are met. rejection due to alteration in fish texture with greater carbon dioxide makes this unsuitable (13). use of high-quality raw fish. For other less fatty marine fish. These conditions include storage of the product at ≤ 3. but is recognised as a drawback of carbon dioxide application. at all times. 5. Only the highest-quality fish. the Sea Fish Industry Authority has published guidelines for the handling of fish packaged in a controlled-atmosphere (16). may result in unacceptable colour and textural change and hence little shelf life extension (11). adequate product labelling in respect of storage temperature.g. Storage studies have shown that the extension in shelf life of MAP cod fillets was proportional to carbon dioxide concentration up to around 50% (V/V). The replacement gas should contain at least 40% carbon dioxide.FISH AND SEAFOOD the safety of MAP and VP refrigerated raw fishery products (15). 4. 2. 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. In the United Kingdom. from both a microbiological and a chemical standpoint. botulinum. The 1985 British Guidelines on MAP/CAP fish from the National Fisheries Institute state that: 1. however. TMAO) that can be reduced to produce 8 . For fatty fish. It was also noted that organoleptic spoilage and rejection by the consumer should occur before the possibility of toxin production. adequate shelf life and cooking requirements.g. The problem of water loss in MAP fish is common. Sensitive and accurate time-temperature indicators must be used to ensure that the product has not been temperature-abused. as already stated.
A. and metabolic activities of microorganisms present in the fish. loss of the fresh characteristics may be simply measured by comparative visual and smell analysis.4. Spoilage of fish starts upon death due to autoxidation (oxidation of unsaturated lipids). but it is a commonly held view that loss of freshness is related to autolytic degradation and spoilage is more microbial in origin (1). to brown and slimy with an intense off-odour. The eyes turn from convex and clear to concave and opaque. reactions caused by activities of the fish’s own enzymes. with no smell. Gas mixtures of 40% CO2/30% N2/30% O2 for white fish and 40 . B. Taste panel evaluation of cooked fish may be used to score or grade colour. and this pattern is the basis of freshness grading schemes. 18) and are probably the most widely used. Loss of freshness and spoilage cannot be separated as processes.4 1.60% carbon dioxide with a balance of nitrogen for fatty fish have been recommended (16. Various methods for whole fish freshness evaluation by a trained panel have been proposed. or for that matter evaluation of the taste of the fish.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.III. Degradation of whole fresh fish stored in ice generally follows a set pattern. these are summarised in Table 1. When working with fillets. 1. and the flesh turns from bright and elastic to dull and soft (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. 17. TABLE 1.CHILLED AND FROZEN RAW FISH off-odours. smell and 9 .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). Over time. so lower carbon dioxide concentrations may have better preservative effects. texture. the gills from pink and shiny. evaluation of taste is unavoidable as the visual pattern of degradation of the eyes and gills is unavailable for analysis. C (C=reject) Significantly. as traditional spoilage patterns of the external organs are typically very distinct.
After death. an anti-freeze. the Eh of fish muscle remains relatively high (20). bacterial reduction of TMAO to TMA.g. spoiled and putrid. cabbage-like. ADP. or its remaining shelf life (1). Generally. TMAO permits a high Eh to remain in the muscle tissue. post-rigor values are typically in the range of pH 5. The process of ATP breakdown is used as an indicator of fish freshness (24). AMP. thus. a ratio of concentration of the hypoxanthine (Hx) and inosine (HxR) to total concentration (ATP. rigor resolves. The pH of the muscle will drop after death depending upon the amount of residual glucose or glycogen that is reduced anaerobically to lactate with the coproduction of ATP. which increases from 0% towards 100% with time.8 to 6. During the initial storage. ii) Neutral taste. an intense odour compound. 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. 23). the cooked analysis of fish passes through four phases of spoilage: i) Delicate sweet. chemical. Because fish tend to have relatively little residual glycogen compared with mammals. After some time. sulphurous and/or rancid smells.5 (18. RT Freshmeter or Fishtester). HxR and Hx) gives a quantity (K-value. little flavour. the muscles relax again and the fish returns to a flaccid state. To a varying extent. However. rigor mortis is the first noticeable change in the fish. possibly slightly metallic. texture becomes soft and watery or hard and dry. 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. an osmoregulator. which has been ascribed a number of possible functions as: a trimethylamine (TMA) detoxified waste product. or simply a waste product present due to bioaccumulation (22). 19). development of sickly sweet. After death. the conductance generally increases. the pH drop of the muscle is correspondingly less. as little endogenous reduction of TMAO occurs (18). reduction in ATP and increase 10 . the muscles harden as residual adenosine triphosphate (ATP) is reduced and the myosin and actin filaments bind to form actomyosin (18). iv) Enhancement of the spoilage characteristic of phase iii. biochemical and microbiological actions. is significant and may even be responsible for the ultimate sensory rejection of fresh cod and other fish with high initial TMAO content (20. These processes are strongly influenced by the physical conditions of storage. By measuring the concentrations of the six components. all marine fish use/have TMAO (21). %). As the fish degrades. The processes of degradation being analysed by the quality scoring methods above are a complex mix of physical.FISH AND SEAFOOD taste of cooked fish. the Torrymeter. From being flaccid. sea-weedy taste. IMP. fruity and/or bitter off-flavours. iii) Traces of sour. ammoniacal. this will generally correlate with the length and severity of rigor.
so the measurement is effective only during the initial loss of freshness. as significant proteolysis is observed only during the latest stages of spoilage and after rejection (30. Lipid oxidation can be a result of enzymic action or a cascade reaction initiated by free radicals (26. 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. Quality indices based upon the products of microbial metabolism do not explain changes in quality until microbial growth produces measurable changes in the fish. 11 . the importance of microbial action increases with time (5. not to describe freshness. the residual acid is then titrated. using Conway diffusion chambers (29) allows determination of changes of mg-N/100 g fish. 7). as microbial spoilage is limited by the low storage temperature (1. colour changes. and are especially important in the spoilage of frozen fish. However. however. a correlation can be made between the spoilage of the fresh fish and the production of TVB-N. or a specific fraction of the volatile bases. 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. The major spoilage odours and flavours of fresh fish are undoubtedly principally microbial in origin. Lipid oxidation and other oxidative changes lead to oxidative rancidity. hypoxanthine. During spoilage. 26). Comparing results for different fish species. Oxidative rancidity is known to reduce the quality of fatty fish in particular (19. which may also be formed by bacteria (25). Evaluation of Total Volatile Base-Nitrogen (TVB-N. 31). 18). This measure is highly dependent upon the temperature of storage and may not reflect the rate of loss of quality equally at different temperatures. Neither does the change in pH during storage correlate well with the production of TVB-N.CHILLED AND FROZEN RAW FISH in hypoxanthine by endogenous enzymes allow this measurement to be used for determining freshness (19). Volatile bases are the best-characterised chemical indicators of fresh fish spoilage. for example the TMA fraction. is later significantly reduced by bacterial action. The Conway method (and variations of it) uses a strong inorganic base to volatilise the bases in the fish sample. does not show correlation between muscle pH and the amount of volatile bases contained within the fish at rejection. also termed TVN). however. 27) produced by aerobic respiration and other forms of metal ion reduction (28). the majority of volatile bases are produced from the soluble non-protein nitrogen of the fish (free amino acids and other low-molecular-weight nitrogenous compounds). these measurements are usually used to quantify the amount of spoilage. Chemical degradation continues after the initial post mortem phase. therefore. For some fish species. and a segregated weak acid to absorb them.
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). 1. The sum of these changes may result in a different spoilage profile. The application of ice storage increases the shelf life of fresh fish from a matter of hours at ambient temperatures to days or weeks. The factors are interactive and cannot be completely isolated.FISH AND SEAFOOD 1. All chemical reaction rate kinetics (and thereby microbial growth) are temperature-dependent. by altering the gas atmosphere (extrinsic). the pH of the fish muscle may change (intrinsic) and a different microbial population may develop due to the change in atmosphere and pH. 5). the lower the temperature of storage.5. Fish spoil as a result of the chemical. The reasons for the differences between tropical. the slower the spoilage processes proceed (within limits). biochemical and microbiological reactions taking place within and on the fish. For fresh fish.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. This increase has been reported to be moderately dependent on the temperature of the sea from which the fish are taken (33. For example. 1. For fresh fish iced immediately after capture and continually stored in ice. 34). a minority are often associated with the majority of the spoilage.and temperate-water fish spoilage rates may be many.5.5 1. The concept of specific spoilage organisms (SSOs) is not new. realisation that the bulk of the microbial population on newly caught fish does not cause off-flavours and off-odours stems from work 12 .3 Recognised specific spoilage organisms (SSOs) The degree of spoilage leading to sensory rejection of fish is partly dependent on the perception of the consumer. Also.5. 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 results of altering one parameter may have far-reaching consequences on others. including the ability of the endogenous bacteria to grow at low temperatures and lower endogenous enzyme activity. careful temperature control during storage is not only important in terms of quality loss. Not all the bacteria growing on a food will lead to the production of objectionable characteristics. but also crucial for the assurance of consumer safety.
During the 1970s.1 Pseudomonas spp. processed (e. They are generally characterised as Gram-negative rods. its ability to reduce TMAO to TMA. These organisms are described as potential spoilers. or faster than the rest of the flora of ice-stored fresh marine fish (9). Pseudomonas spp. putrefaciens growth may be used as indicators of spoilage (30).10% of the total flora of fresh fish from temperate marine waters.5. plus other characteristic compounds of the species responsible for spoilage. It is also present in fresh water. 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 importance of Sh.2 Shewanella putrefaciens Sh. it is often isolated as about 1 . the spoilage of a product may be strongly influenced by the conditions under which the product is held. Members of the genus are able to produce pigments. The spoilage of freshwater fish is generally ascribed to the growth of Pseudomonas spp.e. more especially. obligate respiratory bacteria. on fish are diverse and in many cases species-specific. work with inoculated sterile fish blocks resulted in identification of a specific minority of microorganisms.5. The spoilage compounds associated with the growth of psychrotrophic Pseudomonas spp. mediated spoilage is characterised by ‘fruity’. putrefaciens spoilage of fish is due to its biochemical action on muscle. The Pseudomonadaceae family represent a large and poorly defined group of microorganisms. form methylmercaptane (CH3SH) and dimethylsuphide ((CH3)2S) from methionine and produce hypoxanthine (Hx) from inosine monophosphate (IMP) or inosine. 36). the characterisation of spoilage of each product must be made before the identification of the responsible agent(s) can proceed. and may play some role in the spoilage of freshwater fish. (9) and they are considered SSO of iced freshwater fish (35). putrefaciens to the spoilage of fresh marine fish has been recognised since the 1940s.g. aldehydes.3. which produced the characteristic spoilage compounds of the fish (30.CHILLED AND FROZEN RAW FISH started in the 1940s. although only since the late 1960s has it been realised that specific metabolites of Sh. 1. Bacteria identified as being associated with the spoilage process of fresh fish are as follows: 1. motile with polar flagella. therefore.3. It is thus an important spoilage organism of gadoid fish such as cod. catalase-positive. produce hydrogen sulphide (H2S) from cysteine. and proteolytic and lipolytic enzymes that may affect the quality of fresh and. frozen) fish products. ‘oniony’ and ‘faecal’ odours from the production of ketones. 37). 13 . As previously noted. 35). putrefaciens is considered an SSO of temperate-water marine fish species stored in ice. 35. It is able to grow as fast as. i. Sh. esters and nonhydrogen sulphide sulphur-containing compounds such as methyl sulphide (36. oxidase-positive.
mesophilic strains of Sh. putrefaciens were required. phosphoreum were required for 50% of taste panellists to reject a sample.5. The ferric reductase activity of Sh. and recent taxonomic description using modern molecular methods bifurcated this species. it is strictly respiratory. additional psychrotrophic species such as Shewanalla baltica. 1. Later. putrefaciens strains are characterised as microaerophiles or anaerobes. the bacteria have a significant problem to overcome . owing to its cell size and activity. they are heterogeneous. putrefaciens were identified as Shewanella algae. 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. baltica has been identified as the main H2S producer in cod during cold storing (37). Shewanella oneidensis. It is also able to produce hydrogen sulphide and a range of other off-odour compounds. This wide-ranging respiratory capability is thought to be unique (39). although. In fact. but does show strong chemotaxis up gradients of most of their alternative electron acceptors (39). Shewanella frigidimarina. phosphoreum increased in notoriety when it was proposed that reduction of TMAO to TMA limited the shelf life of MAP cod fillets (12). The localisation of the reductase on the outer cellular membrane offers the bacteria a solution to this problem (40). Shewanella gelidimarina. Ph. Shewanella olleyana. Shewanella denitrificans. and Shewanella profunda were described. Shewanella waksmanii.3. Ph. whereas >108 cfu/g Sh. 41). putrefaciens has been studied. a population of Ph. but Sh. unlike many other motile bacteria. Because no other TMAO-reducing bacteria were present in sufficient numbers to produce the quantities of TMA that were related to rejection. In terms of its taxonomy. putrefaciens growth has been collected (12. 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. does not show carbon-source chemotaxis.FISH AND SEAFOOD for which the most compelling evidence showing spoilage as a result of Sh. In 50% N2/50% CO2 MAP cod. phosphoreum has also been identified as responsible for histamine fish poisoning. as the ferric iron is insoluble. Shewanella affinis. Sh. with several new species being described. putrefaciens lies in its chemotaxis: the bacterium.3 Photobacterium phosphoreum Recognised for some time as being present on spoiling fish (30). Shewanella livingstonensis. like all members of the Pseudomonadaceae. Sh. 41). Initially. Research has shown that approximately 107 cfu/g of Ph. Sh. This type of intoxication occurs when bacteria convert the histidine 14 . phosphoreum sufficient to cause spoilage was found. The prevalence of the bacterium in so many environments. at the time of rejection. 38). putrefaciens was not present in such numbers (23. it was proposed that this organism. Another distinct feature of the physiology of Sh.how to reduce a molecule that they are not able to take into their cells efficiently. and Shewanella aquimarina.
The remaining few bacteria present a major risk involved with the consumption of raw seafood such as sushi or oysters. Fewer still are capable of growing on fresh fish. E. thermosphacta on spoiling fish in a 40% CO2/30% N2/30% O2 MAP (42). (and very rarely F and G) have been reported to cause human disease. types B.3 °C (Group II) and these are non-proteolytic (i. phosphoreum strains have great capacity as histamine producers even under refrigeration conditions. F and G) is well documented (44). Of these. thermosphacta in the spoilage of some MAP fish. 5). Types A. E. this is probably due to spoilage occurring before toxin elaboration. age. B. However.e. presents a potential risk. botulinum recorded up to 1963. When inoculated into VP corned beef and sliced ham. D. Of 404 intoxication outbreaks of type E C. but with proper cooking. Recent studies have investigated the dominance of B.CHILLED AND FROZEN RAW FISH present in fish into histamine. thermosphacta is a well-characterised psychrophilic spoilage organism of meat. Huss (1. Adult disease is caused by the production of the botulinum toxin. algal toxins.3. C1. they do not produce significant product spoilage). it is bacterial risks that increase after capture of the fish. a convenient but diverse species of bacterium with a number of different types. Review of outbreak data suggests fresh fish to be safe. human viruses.1 Clostridium botulinum C. Growing evidence suggests a role for B. The heterogeneity of the types (A.4 Brochothrix thermosphacta and lactic acid bacteria B. MAP studies have also demonstrated this organism’s sensitivity to oxygen and have shown that they are also inhibited by high CO2 concentration (43). B. including antisera. these risks are substantially reduced (1. lightly preserved fish products are associated with botulism. where a particular 15 . a neurotoxin that causes flaccid paralysis and is associated with a variable (44) mortality depending upon dose.6. thermosphacta did not produce off-flavours until 2 . B. Some Ph. C2.3 days after having reached 108 cfu/g (42). botulinum. Acetate production has been reported as a good indicator of spoilage by this organism. 5. 1. previous exposure and access to supportive treatment. 5) reports that fresh fish consumption has never been shown to cause human botulism.6 Pathogens: Growth and Survival Endogenous chemicals. bacteria and higher parasites all present some risk associated with fish consumption (1. 1. especially type E botulinum intoxication. There are few human bacterial pathogens that can cause primary infections or disease and are capable of persisting in the aquatic environment. 1. E and F include bacteria with minimum growth temperatures of approximately 3.5. 6). 75% (with a 35% mortality rate) occurred in Japan.
again. 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. due to the cultural preference for raw fish dishes. but as fresh fish are often cooked before consumption and the botulinum toxin is heat-labile. I-sushi. This strain has been responsible for 50 to 80% of all V. the risk posed to the consumer of fresh fish. USA. parahaemolyticus O3:K6 is responsible for many of the recent V. Russia. V. parahaemolyticus cases have increased across the world. were associated with finfish between 1978 and 1998 (46). A unique clone of V.g. The risk of toxin formation before apparent spoilage has been studied for some MAP fish (45). smoked. V. is small. salted or pickled) fish have also been associated with botulism. which can cause human gastroenteritis. use of post harvest treatments and/or cooking of fresh seafood will reduce risks. which is believed to be required for virulence. parahaemolyticus is a marine organism. parahaemolyticus infections since 1996 and is referred to as the pandemic strain (47). storage and preparation (11). only shellfish are associated with the disease and no reported cases by the Center for Diseases Control. Human disease caused by the organism has only been observed in conjunction with marine bivalve (mostly oysters) and some crustacean consumption. In Japan. which was probably contaminated from an uncooked food source or an excreting foodhandler. parahaemolyticus in sea water fish and shellfish that could potentially be hazardous to humans. sushi) (5). 1978 . The bacterium causes primary septicaemia. there is no specific guideline that describes a minimum level of V. vulnificus is associated with warm marine and estuarine waters. Since 1996. and North America. there remains only an ‘extremely small’ risk of intoxication if reasonable precautions are taken during handling. a Japanese boiled and semi-dried sardine dish. or cooking at 80 °C for 5 minutes) (44) will substantially reduce risk. France. 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. Generally. accounted for the majority of cases (1. 1. Despite the ubiquitous nature of type E C. the majority of outbreaks are caused by consumption of raw fish products (e.2005). the majority are associated with products not cooked immediately before consumption. and proper cooking (boiling for 1 minute. and when in contact with contaminated water (no CDC reported cases from fish. including epidemics in India. Proper chilling. Proper temperature control (<3. Currently.g. The first report of the organism being a foodborne agent was with shirasu.2 Vibrio parahaemolyticus and other vibrios V. Other lightly or semi-preserved (e. especially in individuals with underlying diseases (patients suffer from immune and liver diseases or blood 16 . 11). parahaemolyticus outbreaks. Southeast Asia. whether stored aerobically or in modified-atmospheres. 5).6.FISH AND SEAFOOD fermented raw fish product that is consumed uncooked. Only about 1% of marine isolates produce a thermostable haemolysin. Japan.
but the bacterium is known to survive and grow in the shallow marine. Chill storage may not eliminate growth. monocytogenes incidence has been between 0. but proper cooking should substantially reduce risk of infection. It is particularly associated with disease following consumption of raw oysters from warm sewagepolluted waters. and proper cooking will substantially reduce risk.4 Listeria monocytogenes L. is historically associated with faecally contaminated water. 1. They are aquatic organisms with an epidemiology that is yet to be fully understood. in its habitat. L. 5. P. sodium channel blocking and haemolytic activity (48).CHILLED AND FROZEN RAW FISH disorders). shigelloides are encountered. are associated with human diarrhoeal illnesses. in assessing the risk of fishborne infection by P. no more than circumstantial evidence exists linking the consumption of seafood to Aeromonas spp. listeriosis cases have been reportable to the CDC.000 17 .. cytotoxic. The bacterium has been documented as being responsible for a few fishborne outbreaks of gastroenteritis. their potential to produce disease-causing exotoxin in food has not been fully examined (48). There is no risk associated with the consumption of properly chilled and cooked fresh fish. Despite this. Again. though it is a true mesophile with a minimum growth temperature of 8 °C and shows consequential seasonal variation in its environmental isolation.3 Aeromonas Aeromonas spp. monocytogenes has been well documented as a foodborne human pathogen. shigelloides is similar to Aeromonas spp. which is often fatal (>50%). infection. with minimum growth temperatures around 0 °C and optima of 15 . Although the psychrophiles are unable to grow at body temperature. 17). proper chill storage will eliminate any risk associated with this organism. In the US. however. Similar problems as those encountered for Aeromonas spp. and fish and shellfish are probably the major reservoirs for the organism (50). V.20 °C. cholerae O1. coli on isolation media (50). there is no risk from the consumption of properly handled and cooked fresh fish. and especially estuarine environment. are present on fish and can grow to significant numbers during storage (1. possibly due to under-reporting because of their likeness to E. The major virulence factor appears to be the production of toxins with enterotoxic. others are psychrotrophic mesophiles (49). 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.6. Some isolates are true psychrophiles.55 cases per 100. It is known that Aeromonas spp. the causative agent of cholera. Most cases have been sporadic rather than associated to large outbreaks. and especially Aeromonas hydrophila. Since 2000.6. 1.26 to 0.
within high histidine-containing fish (especially members of the Scombridae and Scomberesocidae families) is responsible for an intoxication termed scombroid poisoning (54). but proper cooking will reduce any risk. AIDS patients and patients undergoing immunosuppressive therapy) and a lack of data covering infectious doses. some Vibrio spp. especially pickled fish. Lightly preserved products. but cooking will significantly reduce the risk of infection. but more especially by Morganella morganii. there is a possibility for growth of the organism on chill-stored fish. Klebsiella pneumoniae and Hafnia alvei.. Fortunately..6 Parasites Helminthic parasites can occur extensively in finfish but very few are capable of infecting humans. It is environmentally ubiquitous but its true frequency in the marine environment is not well studied. and Lactobacillus spp. Low-temperature storage of potentially toxigenic (<5 °C) fish at all times is the most effective way to control histamine production (6). Of fish-transmitted human diseases scombroid poisoning is commonly reported in the US (118 outbreaks with 463 cases reported by CDC 1998 .5 Scombroid fish poisoning The production of histamine and other biogenic amines with human immunological activity. Photobacterium spp. Like other toxins. 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. foetuses.6. The organism poses a serious risk in chilled products not cooked before consumption.98 minutes in cod and D60 of 4.FISH AND SEAFOOD persons. Being a psychrotroph. neonates. Clostridium spp.48 minutes in salmon have been reported (53). leads to the production of histamine. Infections are mostly associated with the consumption of raw or 18 . it is not apparent to the consumer and it cannot be destroyed by cooking. full risk analysis is not possible (52). 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. It is regularly isolated from seafoods (51). 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. Owing to the dependence of the virulence of this organism on the immune state of the host (usually requiring lowered cellular immunity. 1. Decarboxylation of histidine by a wide range of bacteria including Enterobacteriacae.2002 (55)) and is also common worldwide. scombroid poisoning is usually a mild intoxication and it is not a significant cause of death. there is a far greater risk of unacceptable histamine levels being attained before sensory rejection. at risk groups .. 1.pregnant women. If fish are not properly refrigerated. 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. D60 of 1. alcoholics.
The most effective control of all bacterial risks is continual chill storage in ice (0 °C) or freezing. long storage under refrigeration or one of the former prior to freezing. which in many cases will eliminate the risk. For fish from inshore or inland waters of doubtful microbiological quality. Assessment and management of seafood safety and quality FAO Fisheries Technical Paper. In summary. No. EC. area or lot (CAC. FAO.IV) (57).CHILLED AND FROZEN RAW FISH mildly processed fish such as sushi. Faecal coliform counts may be used instead of E. An increase of APC to levels in excess of 106 cfu/g is usually indicative of inadequate refrigeration.1997. 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. parahaemolyticus (see Table 1.H. 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.. Gram L. 2003. risk of human disease caused by natural bacterial contamination (not through faecal pollution) of fresh fish is extremely low. including parasites and/or quantity of their toxins / metabolites per unit of mass. parahaemolyticus Staph. volume. Cooking properly immediately before consumption is also an effective way of reducing or eliminating risk of fresh fishborne disease. Criteria should set standards that are attainable by the currently accepted GMP and applied only when there is absolute need for it.. A limited number of nematodes and trematodes found in finfish have also been identified as a cause of human disease (56). especially in warm-water areas and where fish are to be consumed raw. 444. it may be desirable to test for Salmonella and V. coli proposed by the International Commission on Microbiological Specifications for Foods (ICMSF). aureus 10 7 7 2 3 3 19 5 5 5 0 2 2 0 102 103 103 104 . Testing methods should be practical and the enforcement of such criteria should translate as a reduction of potential microbiological risks to consumers. 1.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. 1997) Huss H. coli counts where this method is preferred.7 Published Microbiological Criteria Microbiological criteria for food defines the acceptability of a product or a food lot based on the absence or presence or number of microorganisms. TABLE1. Rome. Ababouch L.
Nychas G.P. 33. Huss H. 334.N. 6. Incidence of foodborne pathogens in European fish. International Journal of Food Science and Technology. 1995.A. 24.R. 91-8. activities and safety. Jeksrud W. 1994. Quality and quality changes in fresh fish. Critical Reviews in Food Science and Nutrition. Davies A. Gram L. Huss H. 3. 14.. A review of modified atmosphere packaging of fish and fishery products – significance of microbial growth. Food and Agriculture Organisation of the United Nations. 1992.A. Rome. Food and Agriculture Organisation of the United Nations. 1992. 2001. Jehanno D. Ed. Reddy N.8 1.R.S. Ashie I. Ahmed F. Food Control. International Journal of Food Microbiology. 2000. Ed. Davey K. 5.H. 12. Control of indigenous pathogenic bacterial in seafood. International Journal of Food Sciences and Techonology. 1993. Review: Assessing and managing risk due to consumption of seafood contaminated with microorganisms.. 36.H. 87121. 19. 4. Codex Alimentarius Commission.. parasites. Food Control. 348. 5 (1). 243-60. Rhodehamel E.. Incidence and growth of psychrotrophic Clostridium botulinum in foods. 5-7.. 20 . Shelf life extension and safety concerns about fresh fishery products packaged under modified atmospheres: a review. Spoilage and shelf life of cod fillets packed in vacuum or modified atmospheres. 12. 121-37. Fletcher R. 1651-60. Davies A. Capell C. Kautter D. 291-304. 12. 11. 8. Dalgaard P.. 37.J. Food and Agriculture Organisation. Food and Agriculture Organisation. Sivertsvik M. 1989. 2003. Food Control. Simpson B. 1996. The role of seafood in bacterial foodborne diseases. 10.R. in FAO Fisheries Technical Paper No. Spoilage and shelf life extension of fresh fish and shell-fish. 1996. Gibbs P.. Assurance of seafood quality.H. Armstrong D. 283-94. International Journal of Food Microbiology. 2.. 2002..E. 1997.H. 67-71. International Journal of Food Microbiology. Huss H. Journal of Food Safety. Gram L. Huss H.J. Italy. Smith J.H. M. and natural toxins. References Huss H.A.E.K. Feldhusen F..T. Codex Alimentarius Commission....J. 9. 1994.K. Rosnes J. Rome. 2. 27. Kirby R. Microbes and Infection. 7.. 87-118. in FAO Fisheries Technical Paper No.FISH AND SEAFOOD 1. 107-27. 13. Code of Practice for Fish and Fishery Products (CAC/RCP 52-2003).R. Microbiological spoilage of fish and fish products. 8 (2)... Theoretical analysis of two hot water cabinet systems for decontamination of sides of beef.
Rome. Khayat A. 42-54. Journal of Applied Bacteriology. Rome. Schwall D. Microbial and biochemical spoilage of foods: an overview. 121-37. 11. Shewan J.. Lancaster. Autolytic changes in fish post mortem. National Advisory Committee on Microbiological Criteria for Foods. March 20.. 26. Davies A. Food Technology. Part I .. 184. Saito T. 1992.E. in Fresh Fish.. Surette M. Leblanc P. 25.. 1983.A.H. 1933.htm Anon. 1995. National Advisory Committee on Microbiological Criteria for Foods. Eds.M. Slade A. 1985. Biochemical Journal.H. Ratnayake W. Gibson D. 22-36.a review.ucdavis. 1996. Vacuum or Modified Atmosphere Packaging for Refrigerated Raw Fishery Products. Letters in Applied Microbiology. The micro-determination of ammonia. 1982. Food and Agricultural Organisation of the United Nations. 24.. Dalgaard P. Non-enzymatic oxidation of seafood lipids. Reginstein J. 33. Guidelines for the Handling of Fish Packed in a Controlled Atmosphere. 1995. 1988..CHILLED AND FROZEN RAW FISH 15. http://seafood. International Journal of Food Microbiology. Sea Fish Industry Authority. Gram L. Biochemical basis of post-mortem nucleotide catabolism in cod (Gadus morhua) and its relationship to spoilage.J. Nature. Martin R. Shelf life extension of fresh fish . Ed.M.E.. 419-29.. Food and Agricultural Organisation of the United Nations. 2007. Flick G. 23. Qualitative and quantitative characterisation of spoilage bacteria from packed fish. 1-18. 19. in Advances in Seafood Biochemistry. 354-8.N. Quality and Quality Changes. Hebard C. 1992. 1959. 21 16. 19-22. Ackman R. Huss H. 21.. Huss H. 1. Hendrie M. Huis in't Veld J. Technomic. ..R. 21. Bacteria active in the spoilage of certain seafoods. 36.J.M. Gill T..C.. Adopted. 20.H. 1995.spoilage of fish. AVI Publishing. Edinburgh. 26.H. Westport. 27.S. 1995. Flick G. 17. 41-50. Occurrence and significance of trimethylamine oxide and its derivatives in fish and shellfish. 130-2.E. Journal of Food Quality. Herbert R. Lipid oxidation in seafood. 22..J.. Yajima T. in Chemistry and Biology of Marine Food Products. 18. 30. 37. 1971. Journal of Agricultural and Food Chemistry.M. Conway E. 1987.J. 28. Børresen T.E. in Fresh Fish. Gill T. Martin R. Martin R. International Journal of Food Microbiology. 305-17. Byrne A. 27.J. 1415. An absorption apparatus for the micro-determination of certain volatile substances. Huss H. Pedrosa-Menabrito A. Flick G. Eds.J.. 34. Ed. 1996. Arai K.J.J. 117-27.G. Chemical composition. International Journal of Food Microbiology. Fate of Aeromonas and Yersinia on modified-atmospherepackaged (MAP) cod and trout. Washington D.. Hebard C. 33.A. Quality and Quality Changes. 29.edu/haccp/compendium/chapt08.E.E. Microbiological spoilage of fish and fish products. Changes in purine nucleotide of red lateral muscle of rainbow trout.. 245-67.
FISH AND SEAFOOD 31. Journal of Bacteriology. The inhibition by CO2 of the growth and metabolism of microorganisms..M. Blackwell Scientific Publications..A. 1985.. Growth and activity of Shewanella putrefaciens isolated from spoiling fish. Identification of Shewanella baltica as the most important H2S-producing species during iced storage of Danish marine fish.. 35. 36. Dalgaard P. Rome. and regulation.. 1994. Annual Review of Microbiology. 6689-97. Rome. Drosinos E. Lee J.. 317. 33. Gram L. Do fish from tropical waters spoil less rapidly in ice than fish from temperate waters? Food Technology in Australia. Role of proteolytic bacteria in fish spoilage..H. 1995. 328-9. Myers C. 323-9. 34. Processes in Pathology and Microbiology. 1996. Journal of Applied Bacteriology. COUNCIL REGULATION (EC) No 2406/96 of 26 November 1996 laying down common marketing standards for certain fishery products http://eurlex. Archiv für Lebensmittelhygiene. Iron and manganese in anaerobic respiration: environmental significance. 42. a dominant microorganism in Mediterranean fresh fish (Sparus aurata) stored under modified atmosphere. International Journal of Food Microbiology. Vogel B. 40. 44. Jørgensen B. physiology. Gram L. Huss H. and an Achromobacter species. 61-6. 33.H.. 3429-38. Gorczyca E. 67.M.S. 62-70. 311-43. 26.B. Fish spoilage bacteria – problems and solutions. Pseudomonas fluorescens. 71. 1973. Applied Microbiology. 38.europa. 41..F. Scanlan R. 22 . Food and Agricultural Organisation of the United Nations. 13. Satomi M. 1992. Nealson K.H. Oxford.. in Fresh Fish. Miller A.. Nychas G. Dixon N. Venkateswaran K. 262-6.do?uri=CONSLEG:1996R2406:20050602:E N:PDF Sumner J. 1984. Myers J. Do Tropical Fish Keep Longer in Ice Than Temperate Fish: The Circumstantial and Definitive Approaches.. 2002. Environmental Biotechnology.R. Cohen D. Huss H. Quality and Quality Changes. Localisation of cytochromes to the outer membrane of anaerobically grown Shewanella putrefaciens. II. Occurrence and importance of proteolytic bacteria as spoilage indicators. 1989. Kell D. Saffarini D. 1984. 2005. 1982.L. 109-36.H..J. The effect of storage temperature on shelf life. 9. FAO Fisheries Report.R. 72-80. Volatile compounds produced in sterile fish muscle (Sebastes melanops) by Pseudomonas putrefaciens. 43.J.eu/LexUriServ/LexUriServ. Sumner J. Brady P. 32. 36. Applied and Environmental Microbiology.. 4. Karnop G. 51-62. Magno-Orejana F. 18-21. Dalgaard P..M. 37. 334.. 39. Ed. 48.E. Italian Journal of Food Science. Brochothrix thermosphacta. Libbey L. Taussig M. 1989. 174.
K.CHILLED AND FROZEN RAW FISH 45. Presence. Microorganisms in Foods 2. 57. 47. International Journal of Food Microbiology. MacKinnon L. 181. 1-34.M. International Journal of Food Microbiology. International Journal of Food Microbiology. 181-96. 69. Thompson S.. Food Technology. Whiting R. Golden D. Griffin P. Histamine fish poisoning revisited.K. Heat resistence of Listeria monocytogenes in vacuum packaged pasteurised fish fillets. Painter J... 1973-1998. International Journal of Food Microbiology. 23 . 49.C.C. Bishop R. 1991. 54. PCR detection of a newly emerged pandemic Vibrio parahaemolyticus O3:K6 pathogen in pure cultures and seeded waters from the Gulf of Mexico.. Oxford. 1661-6. 20. 55... Majeed K. Ljungh A. Myers M. Lynch M. Garcia G. 1991.. 55 (SS10). 51 (4). 12.. Journal of Infectious Diseases. USA. Fish and Fish Products (Microorganisms in Fish Products). Lehane L.and waterborne pathogens. Morbidity and Mortality Weekly Report.. 1-37. Altekruse S.H.L.P.N. 1993.M. Embarek B.M. Use of risk assessment to reduce Listeriosis incidence. 1997.P. E and F in salmon fillets stored under modified atmospheres at low and abused temperatures.. Mac Rae I. Slutsker L.. Vibrio parahaemolyticus Infections in the United States. Olley J. 1998-2002.. Sampling for Microbiological Analysis: Principles and Specific Application. Blackwell Scientific Publications.. The International Commission on Microbiological Specifications for Foods. 85-95. Genigeorgis C. Microbial Ecology of Food Commodities.. Journal of Food Protection.A. Braden C. The International Commission on Microbiological Specifications for Foods.J..H.A. 2000. Bean N. 2003. Ray B. Aeromonas and Plesiomonas as food. 100-3.. 52. 56. 50.W. 172-249. 303-12. International Journal of Food Microbiology. Smith J.. in Microorganisms in Foods 6. Risk of growth and toxin production by Clostridium botulinum non-proteolytic types B. Daniels N.United States. New York.. 330-6. Miller A. 23. Springer. detection and growth of Listeria monocytogenes in seafoods: a review. Kluwer Academic/Plenum Publishers. 1994. Wadström T. 2000. 2005. 1987. 17-34. Embarek B. The International Commission on Microbiological Specifications for Foods.L. 1986. 181-8. 58. Wilson S.. Surveillance for Foodborne-Disease Outbreaks .. Ed. 50. 51. Woodruff R. Jay J. Huss H.. Loessner M. 2194200. Bej A. Hammond R. Applied and Environmental Microbiology. Modern Food Microbiology. 48.. 2006. Lindroth S. Panicker G. 2005. 12. 53... Experimental evidence for toxin production by Aeromonas hydrophila and Aeromonas sobria in a meat extract at low temperatures.K. 46..J.
. 1985.. 13-32. 1984.. Eds.. 741-50.). Sumner J. 317-21. Florida. Mathematical Modelling Used for Evaluation and Prediction of Microbial Fish Spoilage.H. Further Studies on Marine Fish Stored Under Modified Atmosphere Packaging. The Microbiology of Seawater Fish. Estimation of the bacteriological quality of fish by automated conductance measurements.D.. 73-89. 1992.. International Journal of Food Microbiology.M.. Smith G. 1.. in FAO Fisheries Report No. Studies on the microbiological ecology of mackerel stored by the method of partial freezing. Torry Research Station. Journal of Food Protection. Ravn Jørgensen B. 51. Gibson D. Do Tropical Fish Keep Longer in Ice than Temperate Fish: the Circumstantial and Definitive Approaches. Akiba M.H. Journal of Food Science. 15. 62-70. Cho Y. Kitts D. 317 (Suppl. Torry Research Station of the Ministry of Agriculture.E. Villemure G. in Seafood Science and Technology. Taoukis P. 24 . McMeekin T.. Ogden I.FISH AND SEAFOOD 1. Detection of specific spoilage bacteria from fish stored at low (0 °C) and high (20 °C) temperatures. Application of predictive microbiology to assure the quality and safety of fish and fish products. Gram L. Fisheries and Food.C. Labuza T. 1997. Ross T. Picard G.M. 1. Shahidi F. 271-85.P. Huss H. Simard R. Huss H. Houston N. 4. Magno-Orejana F.. 65-72. 1988.A. Trolle G. Jones Y.. 1989.9.D. Dalgaard P. 1. 6. International Journal of Food Microbiology.. 35. Microbiological quality and shelf life prediction of chilled fish..1983. Ed. Rome. 55. Cann D.9 1. Aberdeen. 295-307. Academic Press. Food and Agriculture Organisation.. in Torry Advisory Note No 93.. Technomic Publishers....M.1 Further Reading Processing Anon. Torry Research Station of the Ministry of Agriculture.L.. Handling and processing of scad. 1961. Hobbs G. Bulletin of the Faculty of Fisheries (Hokkaido University). 1984. Changes in microflora and chemical compounds in mackerel stored by partial freezing. Food and Agriculture Organisation of the United Nations.G. Aberdeen. 127-34. Olley J. 1992. Shinano H..H. Prediction of shelf life and safety of minimally processed CAP/MAP chilled foods: a review. Huss H. in Fish as Food. Borgstrom G. Bulk storage of cod fillets and gutted cod (Gadus morhua) under carbon dioxide atmosphere. International Journal of Food Microbiology. Gibson D.9. 1989. Fisheries and Food. International Journal of Food Microbiology. Lancaster.S. 1986. Ed.2 Spoilage Shewan J.
CHILLED AND FROZEN RAW FISH Einarsson H. Predicting the shelf life of cod (Gadus morhua) fillets stored in air and modified atmosphere at temperatures between -4 °C and +16 °C.R.4 Standards and criteria Council Regulation (EC) Number 2406/96 of 26 November 1996 laying down common marketing standards for certain fishery products.3 Pathogens Condon S.. pre-incubation at low temperature and pH on the thermal resistance of Aeromonas hydrophila. International Journal of Food Microbiology. in Quality Assurance in the Fish Industry. Huss H. 59. Huss H. 1987.. 20023. International Journal of Food Microbiology.. 479-88. http://eur-lex. Elsevier Science.H. Liston J. 1997. 137-46.L. Modified atmosphere packaging of fish and fish products.M. Huss H. 322-6.do?uri=CONSLEG:1996R2406: 20050602:EN:PDF Davies A. Jacobsen M. Inhibitory effect against pathogenic and spoilage bacteria of Pseudomonas strains isolated from spoiled and fresh fish.europa. Hall G. 1992. 72. Gram L..J.H. Effect of culture age.. Gram L.. Applied and Environmental Microbiology. Ed. Otero A. 1992. 25 .H.9. Journal of Applied Bacteriology. 1987. 5. 1.eu/LexUriServ/LexUriServ. Blackie Academic & Professional. 4. Sala F. A rapid method for detection of histamine producing bacteria.9. in Fish processing technology.K. Trolle G... 1. Amsterdam. Klausen N. Garcia M. Eds. 1993. London. Detection of specific spoilage bacteria from fish stored at low (0 °C) and high (20 °C) temperatures. 2197-203. 65-72.
g. This is no longer the case with many processed products now available in stores. and crustacea such as shrimp and crabs. seafood (with the exception of some crustaceans) remains uncooked until the point of consumption. are used in chilled and frozen prepared meals since many of the attributes on which quality is assessed are not present or noticeable (e.g. Simon Derrick The Grimsby Institute Humber Seafood Institute Europarc Grimsby DN37 9TZ United Kingdom 2.1 Introduction It was often thought that whilst the highest quality seafood products are processed and sold as whole fish and/ or fresh fillets (e. colour of gills. the exceptions being Sushimi (high quality raw fish). it does not necessarily apply to the toxins that may have been produced. although recent developments in improved supply chains. 27 . The majority of seafood products produced require a cooking stage before consumption. This has the result that any microbes present in the product have the capability to grow (with subsequent spoilage) unless strict temperature controls are applied. due to the early cessation of spoilage. smell etc. emphasising the high quality of the seafood and the gourmet nature of the prepared products. smoked (hot) mackerel. colourings etc. Sushimi tuna). clarity of eyes. 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. Few seafood products are sold ‘ready-to-eat’.). but still acceptable and meeting all safety criteria. Even though heating prior to consumption will eliminate pathogenic microbes.2. The same used to be said for the difference between fresh and frozen products. smoked (cold) salmon. CHILLED AND FROZEN PREPARED FISH PRODUCTS Dr. products of lesser quality. or could be disguised by sauces. 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.
prawns. and are well known for their diet consisting of waste materials. where the preparation has already been carried out. in particular. Frozen fillets may have a surface layer of 28 . and the consumer only has to heat (cook) the product. Marketing-style descriptions and process definitions are presented below to acquaint the reader with the variety and forms of seafood available.2 Definitions The traditional focus of the chilled and frozen fish market has shifted over the last few years. At the same time. This includes all categories of seafood including trout and other freshwater species. rather than frozen. It is sold from wet fish counters on ice or pre-packed in modified atmosphere packaging (MAP). and this has produced a greater availability and diversity of products in the marketplace. the processing industry has developed a wide range of value added products. as they tend to lose colour and sheen after defrosting. often using the simplicity of a microwave oven. shellfish and molluscs. They are typically flown into export markets. Warm-water/reef-caught fish were initially supplied to add colour and variety to wet fish displays. from both wild capture and aquaculture sources. Shelf life on ice. Cold-water fish microbiological specifications are inappropriate for assessing freshness of warm-water fish. etc. The market spans fresh fish counter sales and frozen recipe dishes with fish as the predominant source of protein. Listeria spp. Seafood is the term used descriptively to include all categories of product. 2. including freshwater fish. The expected shelf life attainable for this format is usually up to 8 days.. molluscs. They can be with or without skin and may contain pin-bones although the major skeletal bones are removed during filleting. which can be farmed. and pathogenic Vibrio spp. Microbiological hazards include Salmonella. but should be equivalent to cold-water products. they are usually of a better standard than species such as tilapia. There is also a limited market for filleted material. but these are now finding favour as products in their own right.FISH AND SEAFOOD Recent trends in the market for food and seafood products. Doubts have been raised over the microbiological status and handling practices of reef-caught fish. have shown the increasing importance of convenience foods. Prepared natural fillets are fillets of fish without any additional coating. Wet or chilled/frozen whole fish are whole fresh fish that may or may not have been filleted.. they are usually supplied filleted with head on. and these are increasingly being presented with an ever-expanding range of accompaniments. Nevertheless. or in MAP depends on catch conditions and treatment. 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.
in VP or through mail order.g. a protective glaze may be added to the frozen steak. These products are packaged in separate high-care facilities to minimise contamination with microbes either from the workforce or from raw products. The pâtés are sold from segregated ‘wet fish’ counters as a ready-to-eat product.. Fish steaks can be cut from a natural fillet or loin (e. and typically uses natural smoke passing over the fillets suspended on racks in a kiln (oak chippings. This process in effect cooks the product and reduces or eliminates microbial contaminants. with a minimum residual salt content of 3. They are sold from wet fish counters. sawdust with other woods adjusted for flavour profile and burn duration).CHILLED AND FROZEN PREPARED FISH PRODUCTS frozen water that forms a protective glaze. tuna steaks) or band-sawn from a frozen block (e. Hot-smoked fish are prepared from brined. mackerel. haddock.5 . Added colours can be included with the brine solution to meet specific requirements for products e. which may. 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. 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. A typical cold smoking process may pretreat the fillet by dipping in a 26% brine solution with a retention time of approximately 3 minutes.2. 29 . salmon. swordfish steaks). Cold smoking is essentially a flavouring process. and salmon. but may be increased (up to 1 month) by packaging methods like MAP or VP.g. the ingredients premixed. be flavoured with herb dips or peppercorns. VP products) because of the high salt content acting as a barrier to spoilage. Fresh fillets are sold from wet fish counters. Cold-smoked natural fillets are prepared from a range of cold water fish including herring. resulting in a product that is ready-to-eat without any further cooking. The cold smoking process does not cook the fish or destroy microorganisms so the expected shelf life is not significantly increased from that expected for wet fish fillets. They are then kilnsmoked to a minimum core temperature of 66 ºC for 2 minutes. in addition.5%. The shelf life for MAP fillets is typically up to 5 days. A core temperature of around 27 30 ºC for approximately 100 minutes is a typical processing condition. Some products may be salted for flavour only (e.g. or pre-packed in MAP or vacuum packaging (VP).g. etc. Pâtés are prepared with raw fish. The expected shelf life of these products is up to 8 days (or longer for some chilled. cod). annatto with smoked haddock. Again.5% in the aqueous phase.2 days) longer than that of non-prepacked fish. mackerel. which is approximately 30% (or 1 . and the product then shaped and heat-processed to cook the product before being chilled. or pre-packed frozen with added glaze. The result is fish with a typical residual salt content of 1. natural fillets of fish such as herring.
rice. Scampi are tails of Dublin Bay prawn or langoustine. addition of ingredients such as colours.or a five-stage process. taramasalata. Breaded fillets/steaks and battered fillets are either natural or shaped fillets. haddock. The cakes are usually coated using a three. and seafood mixes. flavours. This ‘catch-all’ category includes seafood salads. and may include molluscs or prawns that may or may not have undergone a minimal heat process during their primary processing. The protein source can be fish fillet or blocks.. etc. e. cephalopods. or cut from a preformed frozen fish blocks into shapes. which may include molluscs. It is often used as a protein supplement.) Raw material is typically fish fillet. salmon) coated and sold as chilled products. Other. pasta or potato. jellied eels.g. e. There is an increasing market for high value fresh or thawed fish (e. Other ingredients may be natural or reconstituted. ‘fingers’ before being coated in batter and/ or breadcrumbs. Natural prawns from cold or warm waters can include those peeled. rice or pasta. The majority are products made from various types of whitefish (cod. They are pasteurised products and treated as high risk raw materials. Surimi is the insoluble protein fraction of fish after washing and processing. mixed whitefish. coated in batter or breadcrumbs. Band-saw dust and fish trimmings from fishblock preparation materials may be used. and formed into round cake shapes. binders. fish nuggets/bites with dips and seafood-based snack products.FISH AND SEAFOOD Seafood recipe meal centres (frozen or chilled) are places where these products are supplied without a source of carbohydrate. frozen blocks or canned fish and may include molluscs or prawns that may or may not have undergone a minimal heat process.g. tuna. it can be used as a seafood analogue. 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. 30 . With further processing. Fish cakes are products produced by mixing fish with potato and herbs. Prawns. with an expected shelf life of around 7 days. and heat-fixed. long-life seafood mixes using sorbate and benzoate as preservatives.g. and in the shell. and may also include additional ingredients such as fresh herbs and spices.
as a result there is an increase in spoilage rate thereby reducing the product shelf life. Recent public opinion has favoured the use of dolphin. fat content. The Food and Drug Administration (FDA) has produced guidelines for handling and processing of tuna. and processing conditions such as dehydration or poor filleting practices. and an increased risk from a potentially unsafe product (especially those that receive a minimal heat process.and turtlefriendly long lining to the less discriminatory netting. After capture and during any subsequent processing. other potentially pathogenic and spoilage bacterial contaminants may be introduced from a wide range of sources including the environment. 31 . Tuna that does not meet the highest of grades based on colour. searing or bar marking).CHILLED AND FROZEN PREPARED FISH PRODUCTS 2. product handlers. and have emphasised the origin and sustainability of the fish as key marketing points.e. whilst gaping is also dependent on a range of other factors that effect the muscle composition. species (Bluefin>Bigeye>Yellowfin>Albacore). sensory assessment of freshness etc. 2). Tuna caught and discarded as unsuitable for sushi or other high-risk uses is often diverted for canning. Grading is based on size of fish (>25 kg). i. see Table 2. which may become a focus of contamination deep within the muscle. remain on the surface of the fish fillet except where “gaping” or separation of the muscle blocks occurs. and pathogenic flora associated with seafood. on the whole. will the saprophytic flora start to invade the surface flesh (1. 6). The concern is that fish may be visibly acceptable but any subsequent washing process will have little effect on removing contaminants. and ingredients added to the final product. is not used for sushi or other high risk (and high value) products.1 Initial Microflora Seafood Other chapters have discussed in some detail the indigenous spoilage. The edible tissue of some species of fish are more susceptible to gaping than others. A primary role in fish spoilage has been assigned to this organism. the microbial flora of seafood is associated with the gills. it may be used for other products or markets depending on the product specification. storage. specifying internal temperature limits for fish and for limiting temperature abuse during handling and grading (1). and the potentially toxic by-products of histamine-producing bacteria (5.I. Frequent rebaiting and prompt removal of caught fish is essential to limit rapid spoilage. or visible damage from gaff marks (4). Dalgaard et al. handling or preparation stage.3 2. For a summary of background information on the hazards associated with fisheries products and ingredients. and his co-workers have described a large celled microorganism found in spoiling fish and identified it as Photobacterium phosphoreum (3). and intestines of the fish. skin.3. Contamination will. including seasonal variation. Only after death. In summary.
2 Non-seafood ingredients The range of ingredients used in the preparation of chilled and frozen prepared seafood products is very diverse. Clostridium perfringens. nuts and fruit Associated microflora Lactobacilli. toxin. by the HACCP team or by a hazard and risk assessment programme. Enterobacteriaceae.I Spoilage and pathogenic microflora associated with common raw materials and ingredients used in prepared frozen and chilled products Raw material Vegetables (Untreated) including fresh herbs. cream. cereus Bacillus licheniformis. cereus Staphylococcus aureus. TABLE 2. Bacillus cereus. staphylococcal toxin. for chilled products. Table 2. product assembly. Staph. Vibrio cholerae.1 Processing and its Effects on the Microflora Selection of raw materials/ingredients It is important that the potential microbial contaminants of all ingredients. and so are the potential microbial contaminants associated with them. Listeria. B.I provides an overview of common raw materials. Bacillus subtilis B. 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. Clostridium botulinum. coryneforms. Staphylococcus. and milk Cheese Water/Ice Bacillus. pseudomonads.3.4 2. Clostridium Lactobacilli. however. moulds Bacillus.4. 32 . moulds. spore-formers. Penicillium. raw materials. mushrooms. Aeromonas. moulds Bacillus Bacillus. cereus B. and. histamine Cryptosporidium. Shigella. In addition. intermediate processing. streptococci. Aspergillus.FISH AND SEAFOOD 2. be considered at an early stage during new product development. coliforms. shelf life determination. anaerobic spore-formers. and associated spoilage and pathogenic microorganisms. 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. Fusarium. Escherichia coli. Pseudomonas. yeasts Lactobacilli. protozoa and viruses B. cereus Carbohydrates Potato flake and powder Rice Pasta Flour Dried herbs and spices Dairy products including butter. Moulds Enterobacteriaceae Salmonella. yeasts Pathogenic microflora/toxins Salmonella. enteric viruses 2. moulds Bacillus.
2.4.4. depending on the size of the fillet 33 .3 Preparation of fish component of chilled and frozen recipe dishes 2.CHILLED AND FROZEN PREPARED FISH PRODUCTS Any preparation stage or process should be suited to the raw material. 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. as a heat process is often required for functional or organoleptic reasons. Effects on the microflora are minimal during tempering (7).2 Raw material tempering Whenever frozen seafood is used as the raw material it is normally tempered before processing.4. there are a number of distinct methods used.5 illustrate basic principles in the preparation of chilled and frozen recipe dishes. up to -5 °C so as to improve the cutting and handling properties during processing. as the full complexity of the processes used for specific products cannot be covered in a general text. this may be an incidental effect. The use of water for rapid thawing of product is not recommended since it not only rapidly thaws the product surface. 2. Intermediate processing and use of chill for storage of work-in-progress will help to optimise available life of perishable components. Figures 2. Where the raw materials are frozen.3. but it also poses a risk of cross-contaminating other products undergoing tempering. and potential opportunities for growth of foodpoisoning bacteria. Band-sawn materials or fish dice are normally promptly further processed. to evenly raise the temperature of the entire frozen product from -20 °C or lower. All have the same intention. For some components. but it is assumed that any band sawing or dice process will introduce heat from blade friction. defined holding time at a chill temperature (0 – 3 °C). Rapid controlled cooling of heat-processed components will help to minimise spoilage. and contamination from equipment to clean fish muscle.1 . or the use of microwave technology.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. line extensions and process reviews. 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. the core temperature. but will also need to ensure that the optimum shelf life can be safely obtained. Generic examples have been used to illustrate only the major points. 2. Examples of tempering techniques include the use of heated air. It is recommended that the principles of HACCP are applied to novel processes.
It is then size-graded. with the coating heat-fixed. after landing. 2.3.4. are placed into plastic boxes and covered in ice to preserve their condition.Shaped Fillets. 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.2.1 Whole-fillet / scampi Scampi (Nephrops norvegicus).1A . chilled or sold chilled on defrost with an expected shelf life of 6 .Scampi.1.1. may not rise above 0 ºC during processing. the scampi is washed in potable water to remove foreign bodies. The processing of three examples types are described in Figure 2. On arrival at the processing plant. Raw material preparation stages for chilled or frozen coated fish products 34 .1B .3. 2.2).FISH AND SEAFOOD processed.Fish Cakes). Any broken pieces or fragments of frozen tail meat are kept for use in the production of a reformed scampi product.7 days.4.1. 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.1C . Frozen cores are usually batter-enrobed. Fig. by mincing and mixing with a binding agent (as described in section 2. The “blown” meat is graded. drained and then refrozen for storage until required for further processing. quickfrozen.1 (2. The scampi meat (tails) is removed intact from the shell using potable water whilst still frozen. 2.
block-sawn dust. fish fingers).4.off-cuts or belly flaps.2 Shaped products This type of product falls into two categories. The remaining dried ingredients are mixed with the fish.7 days. are handled and stored as any other food product. This type of material will be susceptible to contamination by Enterobacteriaceae and possibly Staphylococcus spp.. or sold as a chilled product after thawing. because of its nature. Spoilage of fillet pieces or trimmings will not be appreciably different from that of fillet. This is especially the case with block-sawn dust. from potato flake. that are to be reused for other foods.g. unless it is poorly handled. It is for this reason that all by-products from primary processing. those that use frozen blocks of whole fillets. spoil more rapidly than the dry ingredients. with the coating heat-fixed. Waste fish generated as a by-product of another process will always be susceptible to spoilage and water loss. as the physical structure of the material has been destroyed. The fish component of the cake will. chilled. iii) Fresh/fillet/minced fish block. Waste material produced from this type of processing is often reused in the third category (fish cakes). and water is added to rehydrate the components. but.3. such as aerobic spore formers and Staphylococcus spp. once the dry components have been rehydrated.4. off-cuts and by-products from other processes (maybe minced) are reformatted give a final product that is still 100% fish.CHILLED AND FROZEN PREPARED FISH PRODUCTS 2. or those where the trimmings.3.1.1. pathogens such as Salmonella and Listeria. Shapes may then be cut from the individual slabs using either hydraulic shape cutters or continued cutting with band-saw type blades (e. Fish raw material can be obtained from a number of sources i) By-product of fish shaping or cutting/dicing process .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. 2. more seriously. or promptly processed to avoid further quality loss. Fish cake cores may be batter-enrobed. quickfrozen. 35 . Herbs or seasonings used in the recipe may contribute additional spoilage organisms or. Therefore. ii) Fish-filleting waste . with an expected shelf life of 6 . The mix is shaped and formed ready for packing or for enrobing. additional spoilage organisms may be introduced. 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. material will need to be transferred swiftly to a chiller.
or sold as a chilled product after thawing. Bacillus spp. milk powder and spices. a. e. The product then passes along a wire mesh belt through a ‘waterfall’ enrober that gives an even coating of batter to the entire product. vibrios and other fishassociated spoilage organisms. Breaded or battered fish fillets are fully heat-processed before consumption. i. perfringens could.. as well as ensuring that there is minimal contamination from human processors. with an expected shelf life of 6 . chilled. Natural fillets d. lactobacilli from the milk powder and flour. egg. and to carry flavourings where required.7 days. which is product-dependent and not always required. A large variety of product formats and types can be batter-enrobed. starch.FISH AND SEAFOOD In all of the above examples. with the coating heat-fixed. Batter contains water. raising agents. quickfrozen. some of this initial flora 36 . Spore-forming anaerobes such as C. rapidly multiply if strict temperature control was not maintained.2 illustrates the typical stages of enrobing or breading. The function of this is to improve the adhesion of subsequent coatings. spices. 2. potentially. and if required. Fish products. and possibly Staphylococcus spp. because of the intensive handling required during processing. e. such as Listeria and Vibrio spp. which typically consists of flour. absorb any excess moisture on the surface of the fish portion.g.or fivestage) frozen or chilled. Scampi tails are particularly susceptible to contamination with Enterobacteriaceae. Products are preformed and presented to the process (either three. any excess is removed with an air knife. Control of temperature limits the potential for growth of spoilage organisms and pathogenic bacteria. If a crumb coating is required. flour. from flour. fish cakes Portions or fillets may be batter-enrobed.e. The first stage. Tempered band-sawn materials b. rapid automated processing with the minimum of delays will minimise product spoilage by pseudomonads. a characteristic flora will develop from the batter components.g. and may include milk powder. Over time. 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.4 Battering/enrobing process Figure 2.4. Initially. the crumb is poured onto the product followed by repeat enrobing and crumbing to complete the five-stage process. is the addition of a pre-dust. which can occasionally be isolated from protein-handling processes. rusk or crumb. and Enterobacteriaceae from flour. IQF/pre-formed fillets c.
but it is believed that poor hygiene regarding product and food handlers. aureus and B. but because batter is heat-set. replaced in part by process contaminants and bacterial flora from seafood. contaminants such as Enterobacteriacae can proliferate and develop during the process. The FDA has issued clear guidelines on acceptable 37 . in the case of B.CHILLED AND FROZEN PREPARED FISH PRODUCTS associated with the ingredients will be diluted out.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.2. dry batter mixes are all potential sources. With inadequate temperature control of batter systems. This can be achieved by frequent clean-downs and adequate hygiene. The origin of contamination for the two organisms is often unclear. and. Staph. cereus can proliferate under certain conditions. this is not of great significance to the microbiological standard of the finished product. It is advisable that some control mechanism is employed to prevent toxinforming organisms from multiplying to dangerous levels. Flow diagram outline of a typical three. cereus.2. and both of these can produce heat-stable toxins that can survive a batter heat-fix process. time-limitation or production run temperature control. Fig.
g. Flow diagram outline for recipe dish ingredients preparation 2. to produce a specific product.3 illustrates vegetable processing and treatment for inclusion in chilled and frozen fish recipe dishes.FISH AND SEAFOOD temperature regimes and run times permitted for batter enrobers. canned fish may be used in their preparation owing to it having wider availability and low wastage. 2. 38 . Vegetable quality is an important characteristic often overlooked in the selection of vegetables for frozen and chilled products. A temperature no greater than 10 ºC for 12 hours cumulatively or 21.4. There are two main categories. Figure 2.3. pasta or potato. rice.2.1 Seafood recipe meal centres (frozen or chilled) These products can be defined as the central component of a meal.1 ºC for no more than 3 hours is recommended (1). Fig.5. Well-managed breading and pre-mix systems that are segregated from batter enrobers present little hazard (7). as this could introduce off-flavours and reduce shelf life. For example. and may include molluscs or prawns that may or may not have undergone a minimal heat process during their primary processing.5 Recipe products These products involve the combining of seafood with other food ingredients as part of a recipe. and must not show evidence of disease.4. The protein source can be fish fillet or block. but they are supplied without a source of carbohydrate e. The vegetables must withstand processing.
Washing with a biocide will at best achieve a 2 . 39 . and may include sauce accompaniments. For example. encourage more rapid decay and loss of structural integrity. and reduce spoilage organisms and potential pathogens. coliforms and pseudomonads throughout the mix. frozen block or canned fish. and mixed whitefish). Salmonella typhi. Once vegetables have been blanched. any processing stage. rice or pasta. In an attempt to improve washing and reduce chlorine taint. larger diced pieces allow for easier handling and a reduction in spoilage but are less attractive to the consumer. or breadcrumbs. they should be stored under temperature-controlled conditions. with the accompanying release of additional nutrients by processing. and may include molluscs or prawns that may or may not have undergone a minimal heat process. i. This is a concern for chilled products that may have a shelf presence for up to 6 days including day of manufacture. A number of examples of direct faecal contamination of vegetables exist in the literature (8). 8) will. pastry. but also as components in recipe meals. coli O157. (7. treated as a highrisk ingredient. loss of quality. for example.5.lactobacilli. colour and water will occur. The opportunity for growth of L. 2. and E. Any assumptions regarding the application of good agricultural practice to produce should be made with caution. However. not just as accompaniments. spore-formers.CHILLED AND FROZEN PREPARED FISH PRODUCTS The products will often be presented with vegetable toppings. the use of night soil or incorrectly applied manure or sewage sludge for ground preparation may contaminate produce with a wide range of pathogens. For premium products. For example. monocytogenes should be taken into account when setting shelf life or holding times of vegetable components.3 log reduction in bacterial numbers on the surface of vegetables. if the process is not appropriately controlled. control is more onerous. coryneforms.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. with cases of foodborne disease traced to Listeria monocytogenes.4. will disperse the saprophytic flora of vegetables . and protected from any post-processing contamination. haddock. even those with pH-modified dressings. Obvious signs of vegetable soft rot due to mechanical damage or infection by Erwinia spp. Products must be washed thoroughly to remove pests and soil. Although the processes are claimed to be superior to using unmodified hypochlorite for washing. all-year-round fresh vegetables are preferred. Blanching of vegetables can be an effective means of reducing or removing surface contamination. The presentation of vegetables has a significant effect on the product quality. chopping and mixing of material. However. The majority are products made from various types of whitefish (cod. acidification of hypochlorite and chlorine dioxide have been proposed and recommended as an improved alternative. Raw material is typically fish fillet.e.
cereus are well known and are still a common issue in the UK (9). Sauces or marinades are added to fish to broaden appeal or to accompany components. and then cooled slowly allowing germination and re-growth. if subjected to temperatures between 20 and 40°C. the preparation process must ensure the eradication of all vegetative pathogens from the ingredients. On hydration. such as butter garnish with black pepper. cereus). If the sauce contains modified starches for freeze/chill stability. Spices and herbs added to form toppings along with breadcrumbs or other coating materials will similarly contribute a spore loading (primarily Bacillus spp. cereus. 11). with the range always expanding as new tastes and cuisines are developed. 40 . especially in those products that have a vegetable component or rice accompaniments. 13). rely on the use of microbiologically sound ingredients. Placing such products in a standard chill store to cool down could result in condensation. monocytogenes and other bacteria that may be present on chilled surfaces such as ceilings. 2. where composite meals are heat-treated. should be chilled to <5 °C as quickly as possible using either blast chillers or other equipment specifically designed for this purpose.) to the product (12. or use barrier or hurdle principles to preclude the growth of pathogenic microorganisms. Manufacturers of chilled products with an extended shelf life must recognise the potential for growth and spoilage of psychrotropic strains of B. but temperature-abused pasta can also spoil rapidly by mould growth.g. Sauces can be multi-component preparations or more simple preparations. a heat process in excess of 73 ºC will be required to rehydrate this component fully. spore-formers can proliferate if adequate control of temperature is not maintained. Typically. An incident involving Staph. This is a particular issue especially where products are heat treated to eliminate vegetative cells.6. aureus toxin is well documented (10. can germinate and rapidly multiply. such as high-risk/low-risk barriers and dispensing and/or holding at above 63 ºC. Appropriate handling facilities or equipment will be required to protect the product.1 Sauce manufacture The variety of ingredients and components used in sauce manufacture is extremely wide. B.4. Chilled pasta is becoming more popular than dried pasta as a component in many product lines. Products or dried ingredients contaminated with either spores of aerobic bacteria (e. a process equivalent to at least a 5D Listeria kill will be necessary. Alternatively.4.6 Preparation of non-fish components of chilled and frozen recipe dishes 2. For this reason all products.FISH AND SEAFOOD Food-poisoning incidents traced to rice and caused by B. and an increased risk of contamination from L. Potato toppings made from flake or powder will normally have little associated spoilage flora.
for instance. 41 . 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. Without barriers.herbs.4 and 2. The format uses flavour compounds . suspended in an acidified sauce with an oil component. 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. flour products.4. and handling of processed sauce can be less rigorous than. that of a dairy-based sauce. Acidified or low-pH sauce formats using acetic acid or vinegar normally require a lesser heat process for the equivalent effect. 13). 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.2. spices. respectively.5 illustrate the recipe dish ingredients preparation and assembly and packaging. etc.CHILLED AND FROZEN PREPARED FISH PRODUCTS Fig. Flow diagram outline for recipe dish ingredients preparation Time limitation may minimise the potential for spoilage by process contaminants or at the dispensing unit.
Increased levels of carbon dioxide will also result in a slight decrease in the pH of the surface tissues.4.7.5. 2. unless the temperature is strictly controlled to below 5 °C throughout the supply chain. generally carbon dioxide and nitrogen.4.2.FISH AND SEAFOOD Fig. so replacing the oxygen necessary for aerobic growth of spoilage microorganisms.7 Effects of packaging on chilled products Products with minimal additional ingredients may use packing techniques designed to minimise microbial growth. which may restrict either microbial growth or enzyme activity. These include: 2. Flow diagram outline assembly and packaging of recipe dish 2. as it dissolves in the surface moisture to form a weak acid. the method replaces air in the sealed packaging with a defined mixture of gases.7. 42 .4. botulinum to grow. and tuna loins are packed by placing the product in a plastic pouch and removing all the air. The objective of this method is to deny the aerobic bacteria (both spoilage and pathogens) the oxygen that they need to grow.1 Vacuum packing A range of fresh chilled products including salmon (smoked and fresh portions).2 Modified-atmosphere packaging Originally developed for the red meat industry. The oxygen-free conditions will however allow other anaerobic bacteria such as C.
as this organism is a common contaminant of freshwater fish species (19). Spoilage of this material is not expected to differ greatly from the processes described elsewhere. It is not clear whether the additional contaminants from components or processing will accelerate this typical seafood-type spoilage. a large number of nutritionally non-fastidious spoilage organisms are commonly isolated from seafood. This is considered to be the one major difference between fish and meat products. The literature has good information on spoilage of individual components of recipe meals. The fish component of recipe dishes. Aeromonas spp. both organisms are capable of growth at chill temperatures. the slow development of ammonical and sulphuric ‘off’ odours and flavours(16). (20). in numerical terms. At refrigeration temperatures (5 . However. 43 . Beyond this. except for some crustacea. and Yersinia spp. Vibrionaceae will succeed (17 ). where the protein component is nearly always heat-processed. The case for growth and contamination of products with Aeromonas is stronger. 2. but little on in-pack spoilage.5. advanced spoilage gives further changes in texture and the production of characteristic ‘off-odours’. but a conclusive case for pathogenicity has yet to be presented for Aeromonas spp. but they will also be readily adapted to the nutrient source and conditions. Spoilage will be most readily detected in proteinaceous components of ready meals. but is not limited to. the microflora of interest will be mainly psychrophilic or cold-adapted (14) spoilage organisms such as Pseudomonas spp. initially. Nonetheless. and finally flavour loss. by a loss of texture.10 ºC) Enterobacteriaceae are often found to proliferate and produce a characteristic type of spoilage that includes. at slightly higher temperatures (10 ºC). Shewanella putrefaciens and Ph.. temperature control is important in preserving chilled integrity of ingredients. phosphoreum. have both been suggested as possible spoilage or pathogenic contaminants of chilled recipe meals (18).5 Spoilage During processing. and in influencing the final eating quality of the product. is typically uncooked or at best lightly processed to retain eating characteristics. followed by moisture loss. This is also true of chill storage and the distribution chain. Not only will these organisms have a numerical advantage over other proposed spoilage organisms.CHILLED AND FROZEN PREPARED FISH PRODUCTS 2. and therefore will have a greater opportunity to dominate other spoilage flora numerically over the course of product shelf life (15).1 Chilled storage For chilled products or chilled components of frozen products.
. The conditions and freezing process most favourable for good-quality products will. Bacillus spp. The ACMSF (22) issued clear advice on this topic and has recommended that the shelf life of products without barriers (23) is restricted to less than 10 days. Characterisation of the enzymes has demonstrated residual activity even after a heat process that would destroy vegetative cells (24). The type and nature of the enzymes and their activity will depend on the bacterial species..2 Frozen storage Maintenance of product temperature at <-18 ºC will not permit bacterial growth or enzymic spoilage. Still. For example.FISH AND SEAFOOD 2. By retaining the intrinsic spoilage flora of seafood. which would warn the consumer of the potentially unsafe nature of the product. eliminating nematodes in products that do not have a heat treatment step. released by a pseudomonad with demonstrable activity to 0 ºC and a peak in production at 10 ºC (15). a recent report examining fish gut bacteria from cold waters characterised a proteolytic enzyme.5. for example. These have been proved to impart off-flavours.3 Enzymic spoilage Post-harvest changes in the quality of fish due to enzymic activity and rigor mortis are well known and understood (10). 44 . and Aerobacter spp. however.g. As a note of interest. Products that are developed with an extended shelf life beyond the 6 . producing characteristic off-flavours. within dairy products are well known.5. also be those that have the least effect on the viability of microorganisms (21). Serratia spp. this should also take into account shelf life of product components used in the assembly of the final product. freezing also has a protective role in improving the safety of raw materials by.7 days typical of fish products/recipe meals must consider the potential for growth of non-proteolytic C. the shelf life of products is limited by the natural spoilage process described earlier. botulinum. spoilage should not be considered a purely microbial phenomenon as taints and off-flavours can develop by non-microbial means (e. where the variety of components and ingredients used in manufacture is varied and complex. This also ensures that any post-process bacterial contamination or spore-forming bacteria in the product will not grow. Spoilage can be either a function of bacterial growth or the microbial release of free enzymes into solution. This reinforces the need for prompt processing and temperature control of ingredients and finished products. It is not unreasonable to expect that spoilage bacteria release many extracellular enzymes.. enzymic processes are often ignored or overlooked as insignificant or unimportant because of the rapidity of fish spoilage. However. auto-oxidation of fats/oils). Pseudomonas spp. 2. Other examples of bacterial proteolytic and lipolytic enzymes produced by Alicaligenes spp.
Counts of between 103 .6 Pathogens: Growth and Survival There are only two species of pathogenic bacteria that can truly be said to occur naturally in fish.6. typhi. soil. and many raw or processed foods (26). in some cases. and this issue may be of greater consequence for warmwater. etc. A number of other Bacillus spp. Vibrio spp.6. any risk of C. Therefore. Inappropriate holding times and temperatures will. spices and herbs and vegetable shoots have been reported (12. and Campylobacter are recorded in filter feeding molluscs such as mussels. potato. or a heat shock followed by holding at a temperature suitable for growth. and some effects related to product type and time to toxin production have been noted. This may occur where storage under gas such as carbon dioxide is used to reduce ripening and so prolong storage. Cases quoting contamination of seafood with organisms such as S. botulinum when held in favourable conditions for growth (28) i. air. water. Incidents in such diverse materials as cream. may be of benefit within any risk assessment for chilled and frozen prepared fish products. have been linked with food poisoning and should also be considered. permit rapid growth of B.CHILLED AND FROZEN PREPARED FISH PRODUCTS 2. especially molluscan filter feeders. Germination of spores requires favourable conditions. cereus. along with previous discussion. botulinum type E and Vibrio parahaemolyticus (25). 2. Additional hazards from marine bio-toxins. 45 .e. 2. low oxygen or anaerobic conditions. perfringens contaminating seafood will depend on the source of raw material. are dealt with elsewhere in this book. agricultural run-off or sewage can contaminate in-shore species. 13.2 Clostridium botulinum There are reports of rapid spoilage of vegetables by C. 2. Hazards attributable to the non-seafood components are listed below and this information.3 Clostridium perfringens A study examining C. In addition. perfringens in seafood noted a transient population of spores in the guts of fish feeding adjacent to rivers and sewage outfalls (29).. histamine intoxication.104/ g should be treated with some concern (26).6. with pathogenic microorganisms or viruses. Numerous quoted figures exist in the literature regarding cell counts required before toxin production occurs. farmed or free-living crustacea (30).1 Bacillus cereus This is a common contaminant of dust. 27). These are C.
monocytogenes as a contaminant in foods (31). when introduced or inoculated into sterile components of chilled prepared recipe dishes. infected food handlers. 2.4 Listeria monocytogenes Interest in this organism has been characterised by a number of very serious incidents associated with a variety of product types (27). monocytogenes tends to grow poorly at refrigeration temperatures in seafood (33). 2. 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). However. and will therefore cause “food poisoning” even in a product that is cooked before consumption. 2. at the present time. These include inadequate refrigeration and preparing foods far in advance of planned service. Shigella spp. the restrictions on growth rate seen with seafood will not apply. and inadequate cooking or holding within the temperature growth range (27). These conditions will allow the production of the toxin. but under the appropriate conditions. All the above points should be considered within any risk assessment of the potential for growth and toxin production by Staph. L. 33) due to its ability to grow at low temperatures and form “biofilms” on surfaces. aureus (35).5 Shigella spp.6. monocytogenes has been frequently isolated from fish-processing plants and is thought to be able to colonise this type of environment (32. An acceptance has now developed in the UK of the presence at low levels of L. indigenous or other. which is not affected by any subsequent cooking process. aureus enterotoxins in foods. 46 . It is unclear. if any other potentially pathogenic organisms.6 Staphylococcus aureus There are a number of underlying reasons for the development of Staph. will colonise factory environments in a similar fashion except for possibly Vibrio spp. but.6. the possibility of a sanitiser-resistant biofilm colonising processing environments should be viewed with some concern (32). L.6. Rates of contamination with smoked seafood are much higher than those with unsmoked products. Sources of contamination are many because of the widespread distribution of this organism in the environment.FISH AND SEAFOOD Vegetable raw materials are potentially contaminated through soil conditioning with animal manure. Chilling may help Shigella to survive. and this is believed to be due to a tolerance to brine used in the smoking process (32). growth may occur in foods (26).
Microbial guidelines.III based on IFST Guidelines (39)) provide example guidelines for seafood products. and are a recognised means of providing data on manufacturing standards. such as batter mixes. (36). it is not surprising that Microbiological Standards (defined as mandatory compliance required (37)) are limited to those raw materials in which specific hazards occur. etc. but these may be eliminated by a subsequent heat-treatment stage. 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. the Institute of Food Science and Technology (IFST) (39) or the British Retail Consortium (BRC) (40). pasta. aureus enterotoxins and B. although due to long analysis times required. and as such the monitoring is for verification purposes rather than control. with respect to the number and types of ingredients used. For instance.6. although the only seafood products specifically identified are raw “shelled and shucked products of cooked crustaceans and molluscan shellfish”. The following tables (Table 2. The criteria themselves will have a range of levels that are dependant on the point in the processing or supply chain at which they apply. and their association with similar materials. and not to final products. cereus emetic toxin are often confused owing to similar symptoms. 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. . pastry.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. It should be remembered that 47 . are not mandatory. In the EU these are defined under European Commission (EC) Regulation 2073/2005 (38). 2. Microbiological specifications do not necessarily relate to product safety. Microbial criteria and the monitoring of microbial levels within a product are important for monitoring trends within production systems. the fresh product is often consumed before the results are obtained.CHILLED AND FROZEN PREPARED FISH PRODUCTS Illnesses caused by Staph. 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.II and 2. 2.A dairy-based sauce that includes a non-pasteurised cheese will contribute significant aerobic total counts.7 Published Microbiological Criteria Due to the complexity of many processed food products. as published by for example.
coated fish. batter etc) should also be consulted when establishing the specifications for raw materials and final product. coli GMP* <104 <102 <10 Maximum Product-dependent 104 103 GMP* Not detected in 25 g Not detected in 25 g <102 <20 Not detected in 25 g <50 ppm (<5 mg/100 g) Maximum Not detected in 25 g 103 104 103 102 <50 ppm (<5 mg/100 g) *GMP = Values found immediately after production under good manufacturing conditions. monocytogenes B. TABLE 2. coli GMP* <102 Maximum 104 TABLE 2.III IFST Guidelines for processed foods . parahaemolyticus (warm-water fish) Histamine (scrombroid fish) Indicators and spoilage organisms Organism APC (heat treated) Enterobacteriaceae E.g.FISH AND SEAFOOD the microbial criteria for other products (rice. pasta. fish cakes Storage: Frozen/chilled Use: Re-heated prior to consumption Pathogens and toxins Organism Pathogens GMP* Maximum Criteria for absence not generally applicable Indicators and spoilage organisms Organism E. aureus V. ready meals Storage: Frozen/chilled Use: Re-heated prior to consumption Pathogens and toxins Organism Salmonella spp.fish products. fish fingers.II IFST Guidelines for part-cooked foods – e. cereus Staph. Maximum= Levels at end of stated shelf life 48 . L.
R. Proper processing and handling of raw tuna. 1993. 8. .. Isolation of a Pseudomonas species from fish intestine that produces a protease active at low temperature. Hobbs G. 59 (2). Powers E. 49 4.. Washington D. Oxford.. 1991. London. International outbreak of staphylococcal food poisoning caused by contaminated lasagne. 2. New York. IBH Publishing Co. 24 (5). Martin R.K.. Bacterial histamine production as a function of temperature and time of incubation. 2. Communicable Disease Report. Sakamoto T.L.R. Journal of Hygiene. Microorganisms in Foods 6: Microbial Ecology of Food Commodities. Fish and Fisheries Products: Hazards and Controls Guidance.. Taylor and Francis. 10. 11. 13. Ishizaki K.R. Brown T. 5 (3). 668 -70. 1976. References US Food and Drug Administration. Journal of Food Protection. Shewan J. 167-93. Incidence and levels of Bacillus cereus in processed spices. Tu A. Christiansen T. Hoshino T.L.H. Van Nostrand Reinhold.C. Powers E. 38 (11).E. Fish: microbiological spoilage and safety... 663-87. Taylor S.. 14. Aureli P.M. Collette R.M. Communicable Disease Report. 1.G. 39 (10). Ltd... 1997. 25 (1). Journal of Food Science... Yumoto I. 12. Recent Advances in Food Science. Journal of Milk Food Technology. 1976. Matsuyama H. Eds.T.. 70-2. 3.M. 1986. 5.. Toxin Related Diseases: Poisons Originating from Plants. 7 (24). in Fish Inspection Quality Control and HACCP: A Global Focus. 2005. 67-73. 1997.J. 1311-7. 1991. 7. 373-8.C. Journal of Milk Food Technology. Lawyer R. Importance of Photobacterium phosphoreum in relation to spoilage.J. Microbiology of Marine Food Products. Ward D. 1997. Pathogenic microorganisms associated with fresh produce. International Commission on Microbiological Specifications for Foods. Latt T.. Outbreaks of foodborne illness in humans. Ohgiya S. 2001.C.8 1.. Letters in Applied Microbiology. Huss H. Kumeta H.. Kluwer Academic. Letters in Applied Microbiology.. 47. The bacteriology of fresh and spoiling fish and some related chemical changes. Bartlett C. 1997. Food Science and Technology Today. 166-73... Murrel H. Slavin J. Cambridge. 1962. Microbiology of processed spices. Mejholm O.A. Bartram P. Animals and Spoilage. Beuchat L. 233-7. 1982. London.W. 9. 96... Food and Drug Administration. 204-16.R. Behling A. England and Wales Quarterly Report. Hackney C. 215. Gilbert R.R. Masuoka Y. 6.L. Wienke A. Pvt.CHILLED AND FROZEN PREPARED FISH PRODUCTS An authoritative survey of specifications can be found in Shapton & Shapton (41). 15. Woolaway M. Dalgaard P.
R. in The Microbiological Safety and Quality of Food. Liston J. a foodborne pathogen? Review of the clinical data. 351-72. Trolle G. Anaerobes in relation to foods of plant origin. Keusch A. 18. Morgan D.C. 27. Fishing News Books Ltd. in Advances in Fish Science and Technology. 155-9. Clostridium perfringens in the environment. London. 1996. 25 (2). Halligan A. Report on Vacuum Packing and Associated Processes. Oxford. Glasgow. Speck M.H.. Food Microbiology. Journal of Food Safety. 13 (1). 1992. 41-3. 30.Yersinia. 4. 4 (1). 655-60. Leatherhead Food Publications. 1996.. Aeromonas and Verotoxigenic E.. What problems does the food industry have with spore forming pathogens B. in Anaerobic Bacteria in Habitats Other than Man. 1993.L. Golden D. Applied Microbiology. Blackwell Scientific Publications.C. Blackie Academic Press. . Connel J. 31.. 1987. Modern Food Microbiology. New York. 26. 51-158 Saxby M.A Literature Survey. 50 17...J. Gould G. coli (VTEC) . 59-72.. Food Taints and Off-flavours. Baird-Parker T. 24. Food Control. 1996. Microorganisms in Foods 5: Microbiological Specifications of Food Pathogens. 1990. Microbiology in fishery science. Blackie Academic and Professional. Occurrence of Listeria monocytogenes in fresh water fish farms and fish smoking plants. 19. Gram L. 32. International Journal of Food Microbiology. PHLS Microbiology Digest. 25. Journal of Food Protection. Lund B. Eds. cereus and C.J. 28. Jay M. Anon. 65. Leatherhead. SAB Symposium Series No.. Matches J... 274-86. 22. 1979. Wood L. Loessner M. 34-40. 1994. 1988. 20. 1986. Detection of specific spoilage bacteria from fish stored at low (0 ºC) and high (20 ºC) temperatures. 1992.E. Food Research International. Ray B. Huss H. U.M. Springer. 29.R. Lund B. Advisory Committee on the Microbiological Safety of Foods. Brooks J. Her Majesty’s Stationery Office. perfringens? International Journal of Food Microbiology. 1974. 472-506.C. 2005. 13.. Mead G. Survey of the incidence of Aeromonas and Yersinia species in retail foods. Curran D... The Emerging pathogens .A.J.FISH AND SEAFOOD 16.H. 309-16. Gaithersburg. Andersson A. 21. Effects of freezing and storage on microorganisms in frozen foods: a review. 9. 40 (5). 28 (4).K. Fresh and processed fish and shellfish . 1995. Huss H. Is Aeromonas spp. 11. London. Microbiological guidelines for some ready-to-eat foods sampled at the point of sale: an expert opinion from the Public Health Laboratory Service. Gram L. Ronner U.W. 333-6. 23.J.V.. 28. Jemmi T.M.J.. Barnes E. Granum P. Ed. Aspen Publishers. Walker S. Liston J. Eds. 2000. Leistner L. International Commission on Microbiological Specifications for Foods. 1977.. Food preservation by combined methods.
Incidence and survival of Listeria monocytogenes in ready-to-eat seafood products. Cambridge. 1992.co.A.H. http://www. Wolfe Publishing. Journal of Food Protection. CRC Press. Smith J. Shigella as a foodborne pathogen. 1991. Chilled Food Association. Institute of Food Science and Technology.. 60 (4). 36. 1987. Gloucestershire. Campden & Chorleywood Food Research Association.eu/LexUriServ/ LexUriServ. Phil Voysey. Psychrotrophic Bacteria in Foods: Disease and Spoilage.do?uri=CELEX:02005R2073-20071227:EN:NOT Institute of Food Science and Technology. Principles and Practices for the Safe Processing of Foods. Woodhead Publishing Ltd. London.F. 51 . Foodborne Pathogens an Illustrated Text. Development and Use of Microbiological Criteria for Foods. U.K. U. 39. Microbiological Criteria.org/Resources/Chilled%20Food%20Association/Public%20 Resources/BRC_CFA_Micro_Criteria_Guidance_Ed_1. 788-801.uk/cday/posters2008/npd6. 40.G.L. 38.chilledfood. London.pdf Commission Regulation (EC) No 2073/2005 of 15 November 2005 on Microbiological Criteria for Foodstuffs. 1999. Shapton N. Guidance on the Practical Implementation of EC Regulations on Microbiological Criteria for Foodstuffs. 372-6. 41. 37. Kraft A. Chilled Food Association.campden.pdf Shapton D. Journal of Food Protection. 1998. McCarthy S. 35. 34.europa. http://eur-lex. 2006.A. Evans M. 1997.A. British Retail Consortium.K.. http://www.CHILLED AND FROZEN PREPARED FISH PRODUCTS 33. Varnam A.2. 50 (9).
snails. Jr. O. while 53 . In these forms. the adductors. Pelecypods have little need for locomotion. composed of two valves hinged together dorsally.1 Definitions General features of the mollusc The phylum Mollusca. lobster. oysters and clams can process about 3. scallops. S. Gastropoda. Box 3012 Bothell WA 98041-4421 United States of America 3. Special tracts of cilia move this mucus toward the mouth. The class Pelecypoda includes all of the bivalves such as clams. and scallops. abalones. Pelecypoda. 4). Other species of shellfish such as crabs.6 litres. cockles. where it is eventually swallowed and digested. Scaphopoda. viruses. the shell. the foot is compressed to form a muscular spade for digging and the head is greatly reduced. Bivalves feed by filtering large volumes of water across their gills to obtain oxygen and food. chitons. Food and Drug Administration 22201 23rd Dr. can hold the shells tightly shut against enemies (1). clams. The digestion process requires less than 2 hours in actively feeding adult bivalves (2). and 2. In most species. P. The class having species of economic and public health importance is the Pelecypoda. Plankton and algae form the bulk of their diet from seawater. lying well within the mantle cavity. and shrimp. respectively. cockles and scallops. clams. MOLLUSCAN SHELLFISH Carlos Abeyta. The phylum consists of five classes: Amphineura. squid. Many microorganisms ingested by shellfish survive the digestive process (3.E. In the process of filtering water.S. mussels. The term shellfish in this context is limited to oysters. and Cephalopoda.3. cockles. octopi. Pacific Regional Laboratory Northwest U. Strong muscles. For example.1 3.7 litres of water per hour. oysters. mussels. chemical contaminants and other impurities on the mucus of the gills. whelks. and tooth shells are a group of soft-bodied animals that usually secrete external protective shells. which includes oysters. mussels.1. slugs. completely encloses the body. they also trap bacteria.
I). 3. Clams are benthic and live burrowed in the mud. many species of bivalves are edible (Table 3.North America Saxidomus nuttali Venus mortoni Protothaca thaca Meretrix spp.I Partial list of bivalve molluscan shellfish products in commercial use internationally (5) Common name Clams Soft (shell) clam Pacific/Atlantic North America Pacific/Atlantic North America Pacific . Mercenaria mercenaria Venus mercenaria Mya arenaria Country Scientific nomenclature Hard clam (also used for Quahaug/quahog) Pacific . but maintain contact with the water by means of a siphon tube (5). Mactra sachalinensis Saxidomus giganteus Japan Pacific – North American Saxidomus nuttali Protothaca staminea or Paphia staminea 54 .2 International commercial products Globally. are not as vulnerable to contamination through pollution of their beds as the bivalves. Whatever contaminants are in the water will eventually get into the shellfish. 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.FISH AND SEAFOOD commercially valuable.1. and usually at higher concentrations than found in their habitat. dwelling on the bottom or attached to structures in the water column.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.
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.
Buried clams leave tubes. cherrystone.FISH AND SEAFOOD Mussel Mussel Mussel Scallops Scallop Coquille St. Most clams are only a few centimetres in their maximum dimensions. 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. hard-shell or soft-shell clam. but 56 . mostly marine bivalve molluscs. laterally compressed. The muscular foot is used to burrow in mud or sand. extended above the surface to maintain water currents needed for respiration and feeding. and two symmetrical shells (valves) held closed by two large muscles and joined by a dorsal hinge joint.1 Clam Clam is the common name of a large group of often edible. A mantle tissue encloses the body of the clam. Different species are variously called quahog. and other names. littleneck. geoduck.1. or siphons.2.
after precooking and brining. uncooked meats packed unprocessed. Dried: Hoshigai (Japan) shelled meat. Canned soups include clam chowder. all in cans. uncooked (shucked). trimmed and packed.MOLLUSCAN SHELLFISH the giant clam of the Indian and Pacific oceans may be nearly 130 cm long and weigh 227 kg. Semi-preserved: meats cooked and packed in spiced vinegar. Dried: shelled meats. and no siphon. hermetically sealed. oyster bisque. boiled and then sun dried for 3 to 10 days before packing in boxes (Hong-Kong) Smoked: meats usually canned in edible oil. washed. Fresh: in shell. packed in weak brine. either whole or minced. then heat-processed. the meats are smoked and packed in oil in cans or jars. smoked meats are also canned. Canned: clams are steamed open and the meats removed from the shells. tidal waters or in brackish water in the mouths of rivers. Frozen: shelled meats. Frozen: in shell.2 Oyster Oysters are marine bivalve molluscs. Smoked: meats. or shelled meats. They live free on the bottom or adhere to stones or other objects in shallow water along the seacoasts. or shelled meats. Soups: oyster stew. Canned: meats removed from the shell by steaming. Live: in shell Fresh: in shell or shelled meats. clam madrilène and clam liquor.1. oyster soup.2. having rough and irregularly shaped shells that are closed by a single adductor muscle. 3. uncooked. These molluscs feed on minute plants and animals carried to them by the current. sun-dried. 57 . Oysters lack a foot or have a reduced foot. in cans with either brine or clam liquor. skewered with bamboo sticks. Live: in shells.
Fresh: shelled meats (often only the adductor muscle and the roe are eaten. Canned: cooked meats. and held apart.1. sauces. 3. Fresh: cooked meats. They have filibranch gills. the shell is often characterised with a lustrous nacreous or pearly lining inside their shells.4 Scallop Scallops are marine bivalve molluscs living in shallow waters of protected bays with sandy or muddy bottoms. The common scallop is about 5 cm long. by interlocking tufts of hair-like cilia. in which the individual branches.1. The shells are fluted and round with a broad. mussel in butter sauce. gutted. Live: whole with shells. Internally. brine.FISH AND SEAFOOD 3. flattened ear or wing at the hinge area. The adductor muscle is the only edible part of the animal.2. also with other ingredients. The familiar marine mussel is asymmetrical. are united. They have a radially ribbed shell with the edge undulated and swim by opening and closing the valves. The posterior halves of the valves contain a large adductor muscle that holds the valves closed. cooked meats packed in dry salt for transport. 58 . Salted: cooked meats bottled in brine.2. mussel paste. smouldered and afterwards dried (Japan). Unlike other sessile bivalve molluscs. or packed with jelly. boiled. 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). Bait: used live extensively for baiting fish hooks for line fishing.) Dried: peeled. in mussel liquor. vinegar solution. Semi-preserves: cooked meats with vinegar-acidified brine and spices. with the posterior ends of the shells (valves) broad and rounded.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. or filaments. and the anterior end smaller and pointed. scallops can swim or leap by a series of rapid closings and openings of the valves. scallops possess a single large adductor muscle. Smoked: meats may be canned in edible oil.
The microflora are dependent upon a number of factors such as season. For example sessile and benthic shellfish dwelling on the bottom or burrowed in marine sediment are especially vulnerable to pollutants (sewage and/or wastewater) introduced into the shellfish water beds.2 Initial Microflora The microflora of molluscan shellfish directly reflects the environment from which the shellfish is harvested. The initial microflora of molluscan shellfish is comprised of the natural commensal microorganisms and the microorganisms accumulated from the water during feeding. which affects the temperature. Because molluscan shellfish are filter feeders. also bottled in brine. and the environment. are vectors of disease transmission (8).MOLLUSCAN SHELLFISH Frozen: shelled meats.5 Cockle A marine bivalve mollusc having a shell with convex radially ribbed valves.1. pH. Vinegar cured: bottled in malt vinegar after brining. mostly in oysters. clams and mussels. Salted: meats either lightly or heavily dry salted. depending on length of journey. 3. Primarily inhabits shallow waters near tidal zones where they bury themselves in the sandy or muddy bottoms. 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 . Fresh: meats removed from the shell by boiling. Raw and partially cooked shellfish. Canned: in brine. in sauce. salinity.2. and butter. Whatever contaminants are in the water will eventually get into the shellfish (7). they accumulate pathogenic microorganisms from polluted waters. Cockles range from about 1 to 15 cm in length and have two valves of equal size. Live: in shell. nutrient concentration and pollutants of the water column. Canned: in own juice. 3.
Vibrios.1 and 3. or by 60 . dredges.. Flavobacterium spp.1. Campylobacter jejuni Escherichia coli Hepatitis A Norwalk virus Other viral agents A variety of commensal microflora are found in molluscan shellfish. nutrients. pollution point sources and hydrological events will greatly influence the initial microflora in bivalves. spirochetes of the genus Cristispira and Saprospira. mussels. Water temperatures affect the concentrations of microflora in the water.. 11. Flow diagrams of the major processing steps of molluscs are presented in Figures 3. responding to the environment in which they are harvested. Acinetobacter spp.2. Micrococcus spp.. Alcaligenes spp. 3. The microflora of molluscan shellfish at harvest consists predominantly of Gramnegative rods while the Gram-positives constitute a minor portion of the flora. At harvest..000 /g.3 3. 3. clams. and yeast cells of Rhodotorula rubra and Trichosporon spp. and shucked shellfish that is shellfish. Other microorganisms found in lesser numbers are the Achromobacter spp.1.FISH AND SEAFOOD Aeromonas hydrophila group Staphylococcus aureus Shigella spp. salinity.. the initial flora composition and concentration of bivalves are highly variable. In summary.1. non-B Snow Mountain virus Plesiomonas shigelloides Salmonella spp. Pseudomonas spp.1 Processing description The method of processing shellstock begins with the harvester collecting shellstock by the utilisation of tongs. that have never been cultured on laboratory media. Temperature. There are microorganisms. 13). The largest groups found are the Pseudomonads. from which one or both shells have been removed.3. and cockles The processing of bivalves takes two forms.. molluscan shellfish have an Aerobic Plate Count (APC) of approximately 1000 to 100. Campylobacter coli Bacillus cereus Hepatitis non-A. 12. Enterococci spp. have been observed microscopically in the digestive tract of Eastern and Pacific oysters. whole or in part. They are termed viable non-culturable microorganisms.3. For example. and Aeromonads. mechanical harvesters.1 Processing and its Effects on the Microflora Oysters. Moraxella spp. rakes. shellstock (live) which is shellfish in the shell.. 15). (9.3..1 Shellstock 3. Bacillus spp.. 10. but have never been cultured (14. observed microscopically.
MOLLUSCAN SHELLFISH hand (Figure 3. by truck or boat. usually the boat will be docked by the processing facility with shellstock piled on the deck of the boat or dock.1. wet storage may be used to increase palatability by increasing salt content of shellfish harvested from low salinity waters. Fig. Thirdly. for transportation to the point of sale. or from other sources approved by the State Shellfish Control Agency (SSCA). after harvesting. Generic flow diagram of live shellfish from harvester to point of sale 61 . or immediately placed in refrigerated trucks. usually in open-mesh containers and/or burlap sacks. The shellstock is transported dry.1). 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 may be wet-stored at the processing facility in tanks of clean seawater.3. Shellstock are placed in drycooler storage containers in the facility. There are three primary reasons for wet storage: first. or in bay-approved shellfish-growing waters prior to sale or processing. Secondly. in containers from which they can be retrieved easily. Shellstock are given a wash to reasonably free them of bottom sediments and detritus as soon as practicable. If by boat. wet storage also may be used to de-sand those shellfish species that tend to accumulate sand in their mantles and gills thus making them more palatable. Water used for shellstock washing is from approved growing areas.
Enterobacter. An outbreak of infectious hepatitis in Sweden in 1956. including V. At this temperature the multiplication of faecal coliforms (E. 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.3. Harvest conditions are another factor that influences the growth of bacteria in shellstock. some pathogenic organisms will survive and multiply in shellfish (20. Shellstock should be refrigerated at temperatures at or below 10 ºC (50 ºF). to avoid possible contamination of the shellstock. however. 17). Washing shellstock helps prevent quantities of sediment and bacteria being mixed with. and temperature storage of shellstock to point of sale. The safety of shellfish is predicated by the cleanliness of the growing area waters from which they are harvested. Shellfish in wet storage tanks are similarly subjected to pollution if the tank water is obtained from a polluted source. 24. bivalves tightly close their shells thus trapping water and microflora. the bacterial counts in the meats will increase.1. At harvest. There is a positive relationship between sewage-polluted shellfish and enteric disease transmitted by shellfish (8.FISH AND SEAFOOD 3.2 ºC (45 ºF) or below (19). vulnificus. without adequate refrigeration. The most important factor in controlling bacterial growth in shellstock is temperature. involving 691 cases was attributed to oysters contaminated in a wet storage area (27). shellstock are not refrigerated on the harvest boat.2 Effects of microflora during processing The initial microflora found on or in shellstock is influenced by sources of shellstock. The washing of sediment and detritus soon after harvesting will prevent further contamination. Several studies have established the increase of bacterial counts in shellstock during post harvest period before the shucking process (18. there is evidence that in the absence of adequate refrigeration. Over a period. 21. Klebsiella. and Citrobacter species) and Vibrios. with the exception of A. If long delays occur before reaching the processing facility and/or refrigeration. handling. Shellfish may contain higher levels of pathogens than are found in the water in which they grow. with prolonged storage the microbiological quality of the bivalve decreases. Generally.1. Excessive sediment on the shells and dead shellfish may increase bacterial 62 . 25). Several studies have established that pathogenic Vibrios and other bacteria may be present in marine sediments throughout the year (21. Furthermore. Water used for shellstock washing should be of good sanitary quality. are prevented. 23. Food and Drug Administration (FDA) recommends that potentially hazardous food such as shellfish be held at 7. 16. Oysters will remain alive in the shell for approximately two weeks if kept at the recommended temperatures. metabolic waste accumulates in the shell serving as nutrients for bacterial growth. synthetic seawater for the purpose of de-sanding and salt uptake. shellstock are placed in refrigerated trucks for transport or placed in wet storage. 22). and the sanitary practices applied during harvesting and shipping. hydrophila (18). for a considerable length of time. Generally. and thus contaminating the shucked meat at the point of sale. 26). when it reaches the processing facility.
MOLLUSCAN SHELLFISH loads in the tanks and lead to increased microbial levels in the shellfish during storage. Thus. In the blower-washer.2 Shucked shellfish 3. the knife is used to cut the remaining adductor thus freeing the shellfish (meats). clothing becomes very soiled. shellstock is subjected to contamination when stored on the floor in standing water. and packing shellfish into containers (Figure 3. for the effects of microflora during the processing of shellstock.1. After washing of meats. If shellfish are not reasonably clean at the time of processing. or subjected to splash from foot traffic.2. removal of shellfish (shucking). Shucking and washing of meats under sanitary conditions prevents the proliferation of bacterial populations. Due to the nature of the shucking operation. Another potential risk with the use of wet storage is the commingling of bivalve molluscs with other marine species such as crab. Washing of meats with potable water is accomplished by placing the meats on a skimmer table (the stand-supported. whereas cotton become wet and 63 . Containers are placed in packing boxes for shipping at refrigerated temperatures. and fish. the tank is opened through a drain gate and chute. the meats are in a tank of water and agitated by the introduction of air from the bottom of the tank. Shuckers routinely use either cotton or rubber gloves during the shucking operation.1. It presents a risk of cross-contamination from the non-molluscan animals to the bivalves. 3.3.2 Effects of microflora during processing The same considerations as previously discussed. washing. washing and culling shellfish prior to storage is essential.2). lobsters.1 Processing description The processing of shucked shellfish is limited to washing of shellstock. through which the washed meats are discharged for rinsing and then packed in containers. are applied to the shucking operation of shellfish. 3. Sealed containers are placed in boxes and covered with crushed ice for chilling or refrigeration and transportation. To remove the shellfish attached to the remaining valve. Studies have shown the use of rubber gloves reduces the contamination level. a considerable quantity of the adhering material will be mixed into the shellfish during the shucking process. In addition.1. Under these conditions if the shuckers enter the packing room. thus contributing to high bacterial counts in the final product (28).3. the finished product may become contaminated. 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). Meats are collected in either perforated or non-perforated buckets for delivery to the washing station.2.3. 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.
Fig 3. Shucking buckets and storage containers are kept at adequate height above the floor to prevent contamination from floor splash. and sanitised before each filling. Separate rooms or lockers for storage of clothing. 2. Shucking containers are rinsed clean with running water. cleaned and delivered to the packing room or prechilled and placed in temporary refrigeration (≤ 2 hours) at 7. 64 . aprons. within one hour. and gloves help prevent the contamination of shellfish.2 ºC or less. 3.FISH AND SEAFOOD retains bacteria during the shucking thus increasing the bacterial load in shellfish (29). 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. Shucked meats are thoroughly drained. There is a tendency to store such articles on the shucking benches or in packing rooms. Shucking buckets are completely emptied at the packing room and no overage is returned to the shucker.2. which may lead to contamination of product.
abundant gas production. Much of the work has been done with oysters and clams. The spoilage process in shellfish may be divided into three stages: increase of acidity. 33. As described previously. thus producing acid and gas (38). Pseudomonas. 36). and packing the shellfish in a timely manner. Shucked shellfish are an excellent medium for the growth of bacteria. 35. where there is a breakdown of temperature control or prolonged storage of shellfish. Temperature abuse will result in high bacterial counts. In summary. in oysters.47 to 6. 32).8% (37). skimmers. or inaccessible surfaces are apt to harbour accumulations of organic material in which bacteria may grow. Glycogen is hydrolysed by the initial flora found in shellfish. For example.2 ºC or less. keeping shellfish refrigerated is the most effective method of retarding deterioration and spoilage. 34. blowers and other utensils that have cracked. Earlier studies. Lactobacilli seem to be the predominant microflora in shellfish. 3. Shucked shellfish are held and transported at temperatures of 7. Studies have shown that bacterial growth is significantly reduced at storage temperatures of less than 7. is stored in the bivalve as a source of energy. All equipment coming in contact with the shellfish such as colanders. at the turn of the century. Micrococcus. spoilage will occur. rough. 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. clean gloves. are an excellent medium for growth of bacteria. these include lactobacilli. Achromobacter and Flavobacterium levels decrease during storage. a carbohydrate. showed that organisms such as Achromobacter. 31.MOLLUSCAN SHELLFISH Unacceptable practices in the packing step can affect the microflora of shellfish. Glycogen. Refrigeration and shipping of shucked shellfish in wet ice is highly recommended. and proteolysis (10). Controls that should be applied include using clean packing equipment.2 ºC. streptococci. and Pseudomonas fluorescens are the responsible agents of spoilage in oysters held at various temperatures (10. and that storage in wet ice is the most effective method for refrigeration of shucked meats (30. whereas bacterium such as Pseudomonas. Shellfish. Alcaligenes. 65 . and yeasts. The environment has an effect on the bacterial population found in shellfish. shucking pails. particularly shucked meats.4 Spoilage There is a lack of information in the literature regarding spoilage microorganisms in shellfish. The initial flora found in shellfish is the major factor of spoilage in bivalves. These organisms may play a significant role in the spoilage processes. particularly oysters stored at 7 ºC. This process changes the bacterial profile to microorganisms favouring low pH. Proteus. Flavobacterium. anywhere during the process of shellfish from harvesting to the point of sale. glycogen levels range from 0.
primarily in brackish or saltwater.5 Pathogens: Growth and Survival This section deals with bacterial pathogens frequently present in shellfish. of V. widespread in both polluted and unpolluted waters. V. 3. It is a common marine bacterium. ranging from epidemic cholera to sporadic cases of diarrhoea. The incidence of V. Fortunately.48 hours with abdominal pain. where outbreaks occur regularly. 66 . there is a recent report of low levels of V.5. shellstock and packing of shellfish in jars) (29). 41). headache and fever. V. The majority of the illnesses were associated with the consumption of raw or partially cooked oysters and clams. however.1. diarrhoea. from harvesting to finished product (i. there were over 250 cases of illness on the West Coast of North American extending from California to Canada.000 population/year (46). Descriptions of outbreaks or reports of growth of the organism in shellfish are presented. The ability of the bacterium to cause gastroenteritis is highly correlated with the production of a heat-stable haemolysin (42). and estimates based from a hospital study show less than 0. parahaemolyticus. during processing. only a small percentage of strains in the marine environment are potentially pathogenic (45). They can be introduced to the shellfish from the environment or added during processing. and to the point of sale.FISH AND SEAFOOD 3. although non-hemolytic strains have been associated with illness (43. Between May and September 1997.1 Vibrios Vibrios are Gram-negative facultative rods naturally occurring in the marine water (estuaries. approximately 23 cfu/g. nausea. Diseases caused by Vibrios are enteric in nature. parahaemolyticus from shellfish is low. parahaemolyticus illness is usually a result of eating raw or partial cooked shellfish (8.e. and harvesting of shellfish in the cooler months. parahaemolyticus possibly producing illnesses (40).1 Vibrio parahaemolyticus V. parahaemolyticus causes gastroenteritis lasting 24 . The control of growth and survival of these organisms can be reduced by the use of good manufacturing practices including immediate and proper refrigeration.5.5 cases/100. Major outbreaks are associated with the warmer months. Products involved were from the same processor. prevention of cross-contamination. Sporadic cases are frequent along the coast of the United States. These organisms can be present in the shellfish from the point of harvesting. One of 250 cases reported involved shucked products with low levels. lot and retail outlet. parahaemolyticus multiplies rapidly at temperatures above 20 ºC. and very common in Japan. 3. Large numbers of bacteria must be present to reach potentially infective levels of >105 colony forming units per g (39). 44). coastal areas). This bacterium is present in all the major processing steps of a shellfish plant.
cholerae non-01 is ubiquitous in the estuarine environment. Inadequate refrigeration will accelerate the growth of V. diabetes. Again. and vomiting. leukaemia.8 case/100. 49). High-risk individuals. 23 of 24 major USA-West Coast estuaries were positive for V.II (8. 3. lasting 6 .000 population/year (8. The widespread distribution of this bacterium is comparable to other studies in the Gulf and Atlantic Coasts (52. The organism attaches to the intestine and release of a toxin is suspected. vulnificus causing septicaemia and death has been associated with the consumption of oysters (8. chills. or immunosuppression. The occurrence of V. with a fatality rate of 50% for individuals with septicaemia. In one study.4 . the organism has been found in the Gulf of Mexico. cholerae non-01 has been associated with septicaemia. those who have liver disease. The bacterium is widespread in estuarine waters particularly in warm waters of >20 °C. Onset of symptoms occurs in 48 hours and can be severe. primarily in immunocompromised individuals with a mortality rate exceeding 50% (50). vulnificus. or primary septicaemia. 57. in the range of 0.0. and preventing cross contamination once the product is cooked. cholerae non-01 is a common bacterial cause of molluscan shellfishassociated illness. 53. fever. Oysters are the most hazardous food associated with V. Control of oyster-associated 67 . the organism has also been found in cooler temperatures from estuaries of Washington and Oregon (47). cholerae non-01 (51). It causes diarrhoea.1. parahaemolyticus growth in shellfish meats is temperature dependent. Control of V. V. vulnificus is by rapid refrigeration during warm weather months. Healthy individuals are susceptible to gastroenteritis. 56). cholerae non-01 is generally a less severe gastroenteritis than that caused by V. parahaemolyticus. V. The bacterium is an estuarine species that causes wound infections. however. 58). and rapidly become inactive at low storage temperatures.3 Vibrio cholerae non-01 Illness caused by V.5. Hazards from this bacterium can be controlled by thorough cooking of shellfish.MOLLUSCAN SHELLFISH Control of V. gastroenteritis. 46. Primarily. which have been implicated in shellfish-related disease outbreaks (31). like V. 55. cirrhosis.2 Vibrio vulnificus The symptoms of V. nausea. parahaemolyticus the estimated annual incidence rate is very low. 3. V. 46.7 days. and nausea within 24 .1. abdominal cramps. vulnificus are fever. 48).48 hours of onset. It is one of the most severe foodborne infectious diseases. Illnesses are usually associated with the consumption of raw oysters as described in Table 3. cholerae. Death has been reported to occur within 36 hours from onset. All isolates are considered pathogenic. these individuals are advised not to eat raw shellfish. They are sensitive to cold. are particularly susceptible to primary septicaemia. 54.5.
63).10% carbon dioxide for optimal growth conditions. 61. V. salinity.10 days. acidity and heat. (8.48 hours with abdominal pain. cholerae non-01 illnesses is by cooking seafood thoroughly. mimicus. Gulf coast environment. e. vulnificus. 62. Poor sanitation and contaminated water supplies will spread the disease. and harvesting of oysters in colder months when Vibrio counts in water are the lowest (59). They are common marine bacteria. 3. Control of infection from these organisms is no different from the other Vibrios. headache and fever.5 Other Vibrios spp. and southern Asia including India and Bangladesh. In general. V. Serotype Ogawa biotype El Tor and 0139 are responsible for epidemics in South American. preventing crosscontamination of cooked seafood. undercooked. imported product from a country experiencing an epidemic. associated with shellfish-borne illness outbreaks.1. processors should know the product source. parahaemolyticus and V. Illness can include abdominal cramps. cholerae 01 in molluscan shellfish. nausea. The control of V. There are several other Vibrio spp. however. 3. the pathogenic severity is lower than V. In developing countries. There is evidence of strains of V. and shock. diarrhoea. In addition.5.FISH AND SEAFOOD V.5. The frequency of infection of these organisms is no different from other Vibrios. 68 .1. The presence of these strains appears to be in low numbers and non-virulent (59). Onset of illness is from 3 to 5 days after ingestion of contaminated food. Illnesses are associated with the warmer months. Death may occur after severe fluid and electrolyte loss. and nausea. abdominal pain. cholerae 01 establishing in the U. low pH. 60). or cross-contaminated shellfish (57. nausea.5%) and 2 .2 Campylobacter jejuni C. It causes diarrhoea. whereas excellent sanitation facilities in developed countries are responsible for the near eradication of epidemic cholera. It is considered a fragile and sensitive organism to environmental stresses such as oxygen. particularly in oysters. cholerae 01. 3. jejuni requires reduced levels of oxygen (3 . and illness lasts 7 . vomiting. and V. and the transmission of the disease by consumption of raw. hollisae. is the same as previously discussed for the non-01 strains. V. drying. fever. widespread in both polluted and unpolluted waters.5. cholerae 01 is transmitted by faecal contamination of food or water. including V. Onset of disease is 6 hours to 5 days.S.4 Vibrio cholerae 01 and 0139 Symptoms of epidemic cholera vary from mild watery diarrhoea to acute diarrhoea with characteristic rice water stools.g. fluvialis. respectively. these organisms cause gastroenteritis lasting 24 . dehydration.
and have been implicated as the causative agent in illnesses from the consumption of raw oysters and clams (68. Sixteen of 28 participants became ill with campylobacteriosis after ingesting raw clams. whereas at 10 ºC Campylobacter survive only for 5 days (71). To reduce the risk of shellfish-borne campylobacteriosis.a (8) Species Hard clams Oysters Soft Clams Mussels Scallops No. 69. Campylobacter spp. parahaemolyticus. Proper care should be taken if the finished product is to be packaged in modified-atmosphere packaging (MAP). Campylobacter appear to survive well in oysters during storage. hydrophila group (75. Campylobacters do not survive well in the environment due to the organism's lack of resistance to environmental factors. 84. and their immediate proper refrigeration. jejuni does not multiply in shellstock at refrigerated temperatures. 68. from Pacific shellstock oysters purchased from a retail grocery store (74). of Cases 4 362 0 1 0 a Includes V. 78) and P. 72. V.5. V. 77. 85). 82. Recently. 71). The seasonality of 69 . although their role as a pathogen is highly controversial and circumstantial. V. 70. they survive well at refrigerated temperatures. jejuni begins in the harvest of shellstock from approved waters. of Incidents 4 279 0 1 0 No. 83. 80) are ubiquitous in the marine environment. 81. mimicus. however. infection occurred among participants of a firemen's banquet. vulnificus.MOLLUSCAN SHELLFISH TABLE 3. They have been implicated in limited shellfishassociated gastroenteritis cases (8. In one case.5. C. shigelloides (79. jejuni were recovered. This finding is significant in that C. 76. have been isolated from shellfish beds throughout the world (67. hollisae Infective dose varies with host susceptibility and virulence of the strain. the incidence of Campylobacter infection is low from the consumption of molluscan shellfish. ranging from 0. and V. Although. V.II Molluscan shellfish associated outbreaks caused by Vibrio spp. 65. control of C.3.4 to 114 cells. 73).3 Other organisms of concern 3. fluviallis. Human feeding studies suggest that about 400 . C. 3. 66).500 cells/gram of Campylobacter may cause illness (64. jejuni survived for over 20 days in market-shucked oysters stored at 4 ºC. however.1 Aeromonas hydrophila group and Plesiomonas shigelloides A. the organism is being continually inoculated into the environment by waterfowl and farm run-offs and thus eventually contaminating shellfish growing waters. cholerae 01 and non-01.
cholerae V.FISH AND SEAFOOD Aeromonas and Plesiomonas is similar to the Vibrios. 3. coli Salmonella V. Enforcement action is considered on a case-by-case basis taking into account all the factors associated with the specific situation. 2 Source (90).000 to 3. hydrophila group stored at refrigeration temperatures. 87) of the outgrowth of A. vulnificus 1 Acceptable Levels in Molluscan Shellfish <500. parahaemolyticus Staph. after 7 days storage of shucked oysters at a shellfish processors’ cooler kept a 5 ºC Aeromonas counts ranged from 500. They are easily destroyed with mild cooking temperatures. and where indicated are intended to be used for official control purposes. vulnificus in molluscan shellfish. FDA/Administrative Guidelines 3 Source (91). prepared sandwiches. action is considered on a case-by-case basis taking into account all the factors associated with the specific situation. suggesting that these individuals are at higher risk when consuming raw or partially cooked shellfish containing these bacteria. 4 FDA and ISSC recommending establishment of an educational program to alert at risk population of presence of V. In one study.600. aureus C. soy products. being isolated more often during the warmer month.0001 negative for the presence3 Under review4 Source (89). They survive well in shellstock or shucked oyster meats held at refrigeration temperatures.000 MPN per gram1 negative for staphylococcal enterotoxin or when the viable MPN count is <10. dried milk/whey drink powders. 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. non-dairy frozen desserts.000/gram (29).6 Published Microbiological Criteria The published microbiological criteria below are included as a guide. The Agency will consider enforcement action against the shipment of molluscan shellfish if the levels exceed as stated. FDA/ Administrative Guidelines. Test Standard Plate Counts E. Caution should be exercised by individuals with haematological malignancies (88). jejuni V.000 cfu/g1 <230/100 g2 <1000 per gram. Pathogen monitoring of Selected High Risk Foods Domestic including: milk. FDA established action levels or levels of concern for certain microbial pathogens in molluscan shellfish. Control of these organisms in shellfish products is limited. Note: Although molluscan shellfish are not included in the selected high risk food. Similar results were seen by other investigators (86. coli Enterotoxigenic E. 70 .
44. 231-51. 5. 1991. Organisation for Economic Co-operation and Development. Eds..MOLLUSCAN SHELLFISH 3.E. 9. Ward D. 12. 71-6. Fish and Wildlife Service..R. 633-40.K. Effects of harvesting waters and storage conditions on yeast population in shellfish. Jr..N. Pfeifer D.W. 447-52. Rowse A. 755-76.S. 1981. Washington D. 82-7. Lee J.S. Quantitative and qualitative commensal bacterial flora of Crassostrea virginica in Chesapeake Bay.. Infectious diseases associated with molluscan shellfish consumption. 245-7. 105-8. 1960. Microbiology of bivalve molluscan shellfish. 1963.. 16. water and sediment from Galveston Bay. Lovelace T. Villee C. Government Printing Office.C..R. Fleet G.. 64. Clinical Microbiology Review. References The Phylum Mollusca. 2. W. in Fishery Bulletin 1964 No. Bernard F. Wiley-Blackwell. 1975. Hackney C. 17. 1964. 6. Journal of Milk and Food Technology. 544-8. Eds. Occurrence of the spirochaete genus Cristispira in western Canadian marine bivalves. 8.. Philadelphia.. Hood M. Tubiash H. 42.R. Rippey S. USA. Jr. 30. Gill S. American Journal of Hygiene. A typhoid fever epidemic caused by carrier bootlegging oysters. 3.S. Microbiology of shellfish: bacteriological study of the natural flora of pacific oysters (Crassostrea gigas). Saunders Company.R.B. 30. 1984. Colwell R. Cook D. 6. Eliot C. Biology of the oyster in relation to sanitation. 15.E. 8. Liston J. Smith F. Tall B. 10. Proceedings of the National Shellfisheries Association.F. 13. Nauman R. 4. Veliger. 13. 14. Journal of Food Protection.. 58. 1994. 29. Bacterial flora of the market oyster. The American Oyster.D.W. Fate of enteric bacteria in estuarine sediments and oyster faeces.A. 19-34. in Microbiology of Marine Food Products. Ed.H. Galtsoff P. American Journal of Public Health. 1970. U. 33643.R. 1995. 11. Ray S.7 1. 1968. 1973. 36.A.. Applied Microbiology. American Journal of Public Health. 1936. New York.M. 46. 7. Applied Microbiology. Vanderzant C. in General Zoology. Applied and Environmental Microbiology.C. Colwell R. 1926. 1983. 72-8. Viability and release Salmonella charity and Escherichia coli from oyster faeces. 104-9.S. 1982. Walker W. Microbiological characteristics of Dungeness crab (Cancer magister). 7.. Van Nostrand Reinhold. 26. 33-6. Journal of the Mississippi Academy of Sciences. Cook D.A. 1940. Microbial flora and level of Vibrio parahaemolyticus of oysters (Crassostrea virginica). 419-25.J. Applied and Environmental Microbiology. Scanning electron microscopy of Cristispira species in Chesapeake Bay oysters. 71 .. Galtsoff P. Thompson C.K. Schmidt C.L. in Multilingual Dictionary of Fish and Fish Products. Old H.R.
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Annals of Internal Medicine. Journal of Clinical Microbiology.. 4.. Wekell M.M. Journal of Food Protection.G.. Fitzpatrick M. 607-9. A method for isolation of Campylobacter spp...E.H. 11. Phenotypic and genotypic characterisation of urease-positive thermophilic Campylobacter (uptc) isolated in N. in Campylobacter II: Proceedings of the Second International Workshop on Campylobacter Infections. 1996. Eds. Robinson D. 56. 70. Wilson I. Brussels.R. 1987. Campylobacter gastroenteritis associated with raw clams.. 1980.A. (jejuni. S41.A.A. The occurrence of Campylobacter jejuni and Campylobacter coli in Sidney rock oyster. Brenner D. International Journal of Food Microbiology. Dalley E.. Journal of Infectious Disease. Disease from infection with Vibrio mimicus..F. 1995. 73. Matsuda M.. 257.. 256. Wilson I. 101-4. 1983.A. Kaysner. 65. Helicobacters and Related Organisms. Blaser M.. Wilson T.R. Johnston J. 68. Campylobacter jejuni in a Washington State shellfish growing bed associated with illness. 71.R. Griffin M. 66.. Feldman R... Morris G. Isolation of vibrio-like group... 4. coli and lari) from shellfish and the marine environment.. Proudford R.E.. Black R. Kaysner C. 99. 1980.A. Helicobacters and Related Organisms. Jr.S.. Blaser M.M. Kaysner C. Wekell M. Jones D.J. Journal of Rapid Methods and Automation in Microbiology. 67. New York.M.S.A. Levine M. Eds.M.. 72. 1983. British Medical Journal..L. Studies of Campylobacter jejuni infection in volunteers.W.G.D.. Moore J. Part 1 Abstracts: Shellfish as a possible source of thermophilic Campylobacter spp.. 1991. Newell D.M. Microbial Ecology in Health and Disease.G. in Campylobacters. Moore J. C. Blake P.. Jr.E.D. Wilson T..P. 1996. Davis B. 1988. Rowe B. 6-9 September 1983.. New York.M.B. 64.G.A. Incidence and survival of thermophilic campylobacters from shellfish growing water: media evaluation. Murphy P. Ireland.. 13. Ketley J. Plenum Press.. 75 .. from patients with diarrhoea.P. Abeyta C. Jr.M. 472-9.M.G. Wareing D. Alam A. Feldman R.MOLLUSCAN SHELLFISH 62. Infective dose of Campylobacter jejuni in milk.. 80. 1993. Journal of the Medical Society of New Jersey.G. Shandera W. 323-5. Pearson A. Plenum Press.. Deeter F.. Murphy P. 1981.J.K.A.M.R.E. Hughes T. infections in man. a newly recognised Vibrio species.G.A. in Campylobacters. Public Health Laboratory Service. Davis J. Newell D.K. 157. Clements M... 169-71. Hunt J.X. Huq M. 621-4. Skirrows M. Abeyta C. 4. Levine M. London.R. Wareing D. Experimental Campylobacter jejuni infection in humans. Huges T... 69. 63..L. Ketley J.. Austin S.I. Arumugaswamy R. 51-64.K. Abeyta C.. Black R.. Stott R. EF-6.. Eds. Clements M.J. 1584.J.. 282.
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1987. The Microbiological Safety and Quality of Food. Ed. Food and Drug Administration. Fish Inspection. Technomic Publishing Co. Washington D. Slavin J.S.S.S. 3.C.1987. Washington D.. U.S. Gould G. Food and Drug Administration.A. Pittsurgh.Pathogen monitoring of selected high risk foods (FY 88/89). Part I Sanitation of Shellfish Growing Areas. 1995.L. 1997.S..12). Food and Drug Administration.MOLLUSCAN SHELLFISH 88... in National Shellfish Sanitation Program Manual of Operations. Food and Drug Administration.600 (CPG 7119. Program 7303. Does Aeromonas hydrophila preferentially colonise the bowels of patients with hematologic malignancies? Diagnostic Microbiology and Infectious Disease. Food and Drug Administration.E. Chapter 5: Foods. mussels. Food and Drug Administration.C. U. Food and Drug Administration. Gaithersburg. in U...R.. 63-8. Lancaster. 7. Ed. National Academy Press. 89. 1991.S. oysters.H.030. Inc.W. 2000.S. 91. Ed. fresh or frozenadulteration by bacteriological contamination Sec. Lund B. Martin R. in U. U. U. Government Printing Office .C.8 Further Reading Ahmed F. Food and Drug Administration Compliance Program Guidance Manual. Baird-Parker T. Collette R. Burdge D. 90. Washington D.C. Seafood Safety. Smith J.M.. U. U. 77 . Clams. Sherlock C.. and HACCP: A Global Focus. Food and Drug Administration Compliance Policy Guides. 1995.E. Quality Control. Aspen Publishers Inc.S.W. 560. Food and Drug Administration.
0 kg polythene bags. Pandalus borealis (deep-water shrimp).4. with additional water to form a glaze.Ph. and are usually aquatic.5 and 1. lobsters and crabs have ten legs. Crayfish. and Trachypenaeus spp. 79 . peeled product. and available in 0. Shrimp can be purchased in the raw or cooked state in a variety of forms: Entire shrimp..g. FRSPH Ethical Trade and Food Management Group Natural Resources Institute University of Greenwich Central Avenue Chatham Maritime Kent ME4 4TB United Kingdom 4.5 and 1.. packed and frozen in 0. lobsters. Species eaten from tropical waters include Penaeus spp. CRUSTACEAN SHELLFISH Linda Nicolaides. Parapeneopsis spp. (deep-water shrimp). prawns and scampi are temperate species commonly eaten by man and include members of the families Nephrops norvegicus (scampi). They possess a segmented body. the front pair ends in claws.0 kg polythene bags. crabs. Pandalus montagui and Crangon crangon (inshore shrimp). and frozen in 1 and 2. e. Shrimps.5 kilo pack (boxes or bags). with shells and heads on or off are usually packed. Breaded shrimp and scampi are battered and coated in breadcrumbs and sold frozen to the consumer for cooking prior to consumption.. M. Peeled and cooked shrimp are ready-to-eat after controlled thawing. crayfish and shrimp (1).1 Definitions Crustaceans are invertebrate animals and represent a class of the phylum Arthropoda. jointed limbs and a chitinous exoskeleton. Such products are usually only partially cooked before freezing. Peeled ready-to-cook shrimp are raw.
for crabs and lobsters.FISH AND SEAFOOD Crabs. with the increasing demand for ready-to-eat/cook crustacean products. e. storage above the recommended storage temperature. tropical waters. Shewanella putrefaciens and 80 . their feeding and living habits. The structure of the circulatory system of crabs is not closed. Cooked and chilled. the flora will change depending upon the methods of handling and/or the environmental conditions to which they are exposed. so that spoilage may occur during landing and transportation to the processing plant. or frozen crab. Control measures to prevent these potential hazards occurring include training and supervision of operatives as well as Good Hygienic Practices (GHP). the season. Poor hygienic procedures will increase the probability of contamination with bacteria of public health significance. traditionally. 4. which means that the haemolymph can be a reservoir of bacteria. The microbial flora of crustaceans from temperate waters differs from that of crustaceans from warmer. there is a growing market for fresh lobster tails or cooked crabmeat in crab shells. the level and type of the initial microflora of crustacean shellfish will reflect a combination of factors.. lobster and crayfish are available for the consumer owing to the growing demand for ready-to-eat products. In comparison. As with finfish. particularly after it has been cooked. during the peeling of cooked shrimp.g. lobsters and crayfish are traditionally kept alive after capture and then cooked to kill immediately before consumption. e. it is important that shrimp are handled and iced correctly in order to keep them in as fresh and uncontaminated a state as possible. Following capture or harvest. particularly for members of the genus Vibrio (2). spoilage bacteria are of little importance as. which include the environment from which they have been captured or harvested. only live carcasses are processed. shrimp die immediately after capture. stored and transported as frozen product. Spoilage bacteria associated with temperate crustaceans include Pseudomonas spp. Pasteurised crabmeat is edible meat removed from the exoskeleton by hand. and the temperature and quality of the waters in which they exist. However. the potential exists for crosscontamination from both the processor and from the environment.2 Initial Microflora As with finfish.g. Microbiological risks associated with crustacean shellfish increase in proportion to the degree of handling that the product undergoes. the geography of the area they are captured from or are grown in. Spoilage begins following death once the immuno-response system of the crustacean shellfish fails to function. hence. boiled to kill the crustaceans and cook their meat prior to consumption. For example. lobster tails and cooked crabmeat presented decoratively in their shells. If these products are subjected to temperature abuse. packed into polythene bags. this presents more favourable conditions for growth by both spoilage and pathogenic bacteria.
gills and thorax. As the muscles of crustacean shellfish decompose rapidly after death dead animals are rejected. however. In contrast. In comparison shrimps die quickly after capture. 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. When the shrimp arrive at the processing plant they are washed and then sorted into size. The claws are usually bound together to prevent the animals fighting and causing damage to each other. Pseudomonas spp. Micrococcus spp. peeled and cooked. bacterial contamination and spoilage are inevitable.1 Prawns and shrimps An example of the process used for preparing frozen. there is evidence that the shelf life of tropical crustaceans is longer than that of temperate-water species stored correctly in ice . chilled 81 . These animals can survive out of water if the surrounding environment is kept cool and damp. Scrupulous hygiene is important at this stage to prevent the cross-contamination of the shrimp tails. Vibrio spp. 4.3 Processing and its Effects on the Microflora Crabs. thus.. or with the shell and central vein removed. lobsters and crayfish are harvested from cages or pots containing bait.CRUSTACEAN SHELLFISH members of the Acinetobacter-Moraxella group. up to 16 days for tropical species compared with 8 .1). and will be dependent upon whether the crustaceans were harvested from cool or warmer waters.. As with finfish. graded according to size. The latter process can expose the shrimp muscle to incidents of cross-contamination if GHP are not followed. They are then taken to market alive in either holding tanks. chilled sea water. they are washed. from the time of capture and processing. Shrimps can be processed whole. with “head on” or “head off” whereby the edible tail is removed from the head. Shrimp might be further processed with the shell on. After capture and transport to the processing plant.3. 4. by removing the head and thorax a source of enzymes is eliminated that will prevent the occurrence of black spots. and Achromobacter spp. corynebacteria. uncooked prawns and shrimp is presented in Figure 4. the rate of deterioration of the crustacean shellfish by these organisms is reduced. glazed and frozen (see Figure 4. A large proportion of shrimps are produced by aquaculture. The rate of spoilage will be a function of time and temperature. whether GHP are in place. so they are either sold on the local market or are processed in local processing plants.for example.. Shrimp will then be glazed and frozen in a raw state. especially if they are stored in ice or iced water. the genera associated with tropical species include Enterobacteriaceae. or are found in brackish water around the shores.10 days for temperate water species (3). or cooked. or transported directly to processing plants. Hence.1. Alcaligenes.
Good Manufacturing Practice (GMP). 4. Flow diagram .FISH AND SEAFOOD frozen. and packed to meet the customer’s specification. Breaded products introduce an added risk of spore forming bacteria.1. should be followed to ensure that the product is not re-contaminated with microorganisms present in the processing environment or from the workers. including GHP.frozen uncooked prawns 82 . Fig. 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. spices and cooking batter. particularly the pathogens Bacillus cereus and Clostridium perfringens originating in the flour. After cooking.
dull and sticky.3. indicative of protein degradation and the growth of the principal spoilage bacterium Sh. bacteria of public health significance. However. The shell will be soft.2 Cooked crabs and lobsters As indicated earlier. 4. Once removed and cleaned.2 Crabs and lobsters Crabs and lobsters should be alive when boiled or sold. there is no indication of life. they include Sh. there is a new hazard of contamination of the cooked meat by workers during its physical removal from the exoskeleton. bright and dry appearance.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. or in hermatically sealed cans that have undergone an appropriate heat treatment. If boiled. viruses or parasites. the joint between the tail and carapace is open and the limbs hang loosely. contaminating microflora may include enteric bacteria. An offensive odour is also produced. spoiling prawns and shrimps have shells that are soft.4. since crabs and lobsters are traditionally consumed immediately after cooking. With spoiled crabs and lobsters. If hygienic procedures are not followed. with the growing demand for ready-to-eat products from crab and lobster meat. and have a sweetish. 4. If the tail drops. wet and ‘soapy’ or ‘jammy’ to the touch. the meat is either repacked and sold in the shells. In comparison. slightly iodine smell. or packaged and processed. The shells have a crisp. the tightness of limbs will indicate whether they were killed by immersion in boiling water or.4. and a strong ammoniacal odour is produced.4 Spoilage The main genera of Gram-negative bacteria responsible for the spoilage of crustacean shellfish are the same as those that cause spoilage in finfish. 4. 83 . whilst boiled crustaceans have a pleasant slightly sweet smell. putrefaciens and Pseudomonas spp. with visible decomposition or green discoloration seen through the membrane exposed by the dropping tail. Processed crab and lobster meat can also be bought as packages of meat that have been subjected to a pasteurisation step. Heat is produced if the shrimps decompose in a confined space.CRUSTACEAN SHELLFISH 4. putrefaciens. for crabs. there is no associated spoilage problem. immediately prior to boiling.
Aquatic birds and the hands of workers have also been identified as sources of the pathogen.5 4. Salmonella cannot be completely removed from raw shrimp during processing. in compliance with the Food and Drug Admistration’s (FDA) ‘zero tolerance policy’ for L. Further surveys identified Listeria spp. An import alert was put in place in June 1987. which required automatic detention of all Mexican frozen crabmeat that tested positive for Listeria and Escherichia coli (12). They are mesophilic microorganisms usually residing in the intestinal tracts of human beings and warm-blooded animals. in 24. Moreover. shrimps and cockles sold (17). However. The pathogen was later identified in frozen raw shrimp (13) and lobster tails (14) . One of the earliest accounts of the presence of Listeria monocytogenes in crustaceans was by Russian workers in 1959 (9). 6. animal manure. cooked and smoked fish. Salmonella species are more commonly found in crustaceans reared by aquaculture or growing in faecally contaminated brackish water compared with species of marine origin (4. and to increase testing of many other domestically produced seafoods under the General Pathogen Surveillance Programme (14. monocytogenes in cooked ready-to-eat crustaceans is recognised as a potential risk to selected groups of individuals: the very young and 84 . shrimp and shellfish (11). 16). even if GMP is followed. although efficient cooking will quickly eliminate it (6). 7). Results of a survey carried out in England and Wales between 1987 and 1989 demonstrated that L. later surveys in 1990 recovered the pathogen from retail raw.2 Listeria spp. 4. a common pond fertiliser used in aquaculture. monocytogenes in cooked ready-to-eat foods.1 Pathogens: Growth and Survival Salmonella spp. 5. Although foodborne illness had not been associated with the product. often contains Salmonella and other enteric pathogens (8) and has been demonstrated to contaminate crustaceans growing in these environments. The above results prompted the FDA to develop a compliance programme for the pathogen. Following this. monocytogenes was absent from 40 samples of cooked prawns.5% of frozen raw shrimps from different countries (13). as well as for Salmonella. Hence. the first in a series of Class I recalls was issued in May 1987 to retrieve nearly 4 tonnes of tainted crabmeat (11). The genus Salmonella contains a wide variety of ‘species’ pathogenic for man or animals. The presence of L. nearly 30 years elapsed before the potential hazard of L.cooked prior to consumption. and usually for both. in domestic/imported shrimps (15). monocytogenes in contaminated crustacea was recognised in the United States of America (USA) when the pathogen was isolated from Mexican frozen cooked crabmeat (10).5.FISH AND SEAFOOD 4. fish fingers.5.
which can become colonised by V. Crustaceans have been linked with food intoxications attributed to Staph. pregnant women and the immuno-compromised. cholerae non-O1. Vibrio cholerae. the pathogen can grow at temperatures as low as 3 °C.5 Vibrio spp. cholerae O1 and V. e. The pathogen has resistant spores. 19).g. Researchers have demonstrated that inoculated shrimp supported toxin production by C. especially in products that have been vacuum packaged. is widely distributed in aquatic environments. 85 . was associated with seafood consumption. 24. 4. although more recently the non-O1 type has also been demonstrated to cause foodborne illness in crustaceans. which was eventually attributed to a ship discharging its bilge into the coastal waters of Peru. particularly crabmeat (18. therefore.70% of healthy individuals carry toxin-producing strains of Staph. cholerae. combined with the degree of handling of these types of product.5. whilst boils and cuts are also common sources of the pathogen. Crabs are covered in a chitin exoskeleton. pasteurisation and cooking. the causative agent of cholera. monocytogenes has been attributed to crustaceans. The main source of contamination is the hands of processing personnel. 4. as part of the natural bacterial flora.CRUSTACEAN SHELLFISH old. Control should be achieved by promoting GHP and staff training programmes. 25).4 Clostridium botulinum C. V. so is able to survive heat treatments that eliminate vegetative cells.5. V. about 50 .3 Staphylococcus aureus Staph. However. although contamination by types A and B may occur from the environment. Once the competing microflora has been removed by a heat treatment. 4. 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 is not a member of the intrinsic flora associated with crustacean shellfish. 22) as well as from undercooked crustacea when harvested from contaminated waters (23. GMP should be followed in the processing plant to ensure that the final product is held at 3 °C.5. cholerae is divided into two groups. aureus on their hands. aureus because of the ubiquity of this organism in the processing environment. Vibrio parahaemolyticus and Vibrio vulnificus are species that have been reported to be associated with raw crustaceans harvested from estuarine waters (21. To date. botulinum type E at 10 °C but not at 4 °C (20). An epidemic that began in South America in 1991. The former. no incidence of foodborne illness caused by L. botulinum type E is commonly associated with fish and fishery products.
4. 4. hydrophila and A. 4.5. been implicated in a case of foodborne infection associated with crustaceans. Norovirus. a marine halophile. V. hydrophila has been isolated from prawns (29). A.8 Parasites The use of animal excreta in aquaculture has also been associated with foodborne parasitic infections of crustaceans. although A. Parasitic infections are controlled by either cooking infected material prior to consumption. 32). Caliciviruses. Of these.5. It is more commonly implicated in countries where crustacean shellfish are eaten raw. 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. particularly trematodes (6). but has the potential to cause fatal foodborne illness (26). jejuni has not. occurs in crustaceans from warm environments.FISH AND SEAFOOD and caused major economic losses to South American countries that export shrimp to the United States.5. 4. to date.30 MPN/g) (27). Astroviruses and non-A. Japan.9 Viruses Crustaceans grown in waters contaminated with sewage may become carriers of viruses such as Hepatitis A.7 Aeromonas hydrophila and Aeromonas sobria A.6 Campylobacter jejuni and Campylobacter coli C. coli. 31. The organism is widespread in estuarine waters and is a common isolate from harbour water in warm climates. vulnificus is an invasive and lethal pathogen associated with wound infections. however. parahaemolyticus. jejuni was isolated from 36 (15%) of 240 samples of freshly hand-picked blue crab (Callinectes sapidus) meat. V.5. the potential hazard exists as C. However. in a survey carried out in the United States between 1977 and 86 . South America.8% of samples contained C. However. hydrophila is able to grow well at refrigeration temperatures (0 . e. Hepatitis A virus presents the most serious hazard. Quantitative levels were below limits of detection in all cases (<0. non-B Hepatitis viruses (31). sobria are known to be fish pathogens and are commonly isolated from ponds used for aquaculture. they have not been confirmed to be causal agents of foodborne illness associated with crustaceans (28).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. or freezing at a temperature of -23 °C for 7 days (30. whilst 5.g.
Both Hepatitis A virus and Norovirus are stable pathogens and are relatively resistant to inactivation (34). The toxin causes amnesic shellfish poisoning (ASP) when consumed.10 Allergens Allergens have not been reported to be a significant hazard with crustacean shellfish.47% of outbreaks attributed to Hepatitis A virus were attributed to a foodborne origin (33). or chlorine concentrations similar to those found in drinking water. 4. risk-based system based upon the seven principles of Hazard Analysis and Critical Control Point (HACCP) supported by pre-requisite or good practices programmes at all stages of the food chain. Norwalk viruses can also withstand heating to 60 °C. consistent with that in the stomach.1 EU Legislation Microbiological standards for cooked crustaceans and shellfish. which could be transferred to the meat during unskilled removal from the shell (35). from farm to fork (37.5. No foodborne incidents have been attributed to this source to date. produced or imported into the European Union (EU). 4. Selected parts from this regulation is presented below.CRUSTACEAN SHELLFISH 1981. 39).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. which results in a persistent and apparently permanent loss of short-term memory (36).5. although it is one of the European Union (EU) legislated allergens. 4. 38. Therefore any microbiological criteria need to be part of this HACCP-based system to verify that the system is under control. Provisions laid down concerning the use of additives would also apply to these types of products. 2073/2005. They can survive low pH levels. 87 . only 0.6 Published Microbiological Criteria The global legislative requirement for controlling the safety (and quality) of crustacean shellfish is to apply a preventive. The risk of such hazards is far higher with fin fish and molluscan shellfish. are included in the Commission Regulation (EC) No. 4. A summary of microbiological criteria for crustacean shellfish is listed below.6.
FISH AND SEAFOOD
220.127.116.11 Microbiological criteria applicable to the production of cooked crustaceans (Commission Regulation (EC) No 2073/2005 on microbiological criteria for foodstuffs) 18.104.22.168.1 Chapter 1 - Food safety criteria
L. monocytogenes Prepared crustacean products fall into the category described in the regulation as “Ready-to-eat foods available to support the growth of L. monocytogenes other than those intended for infants and for special medical purposes”. During capture and preparation there is a zero tolerance for the presence of L. monocytogenes on the product. However, to take into account the issues of natural habitats being contaminated with low levels of this pathogen there is a slight loosening of the regulation at the point of sale and for the duration of the products shelf life (refer to Table 9.I, Food Category 1.2). If the level of L. monocytogenes should be above this level then the consignment is considered to be unsatisfactory. Salmonella All cooked crustaceans and molluscan shellfish should be free from Salmonella at the point of sale and for the duration of their shelf life (refer to Table 9.I, Food Category 1.16). Should any sample test positive for Salmonella then the product is declared to be unsatisfactory. 22.214.171.124.2 Chapter 2 - Process hygiene criteria
Process hygiene criteria for E. coli and coagulase-positive staphylococci in shellfish and shucked products of cooked crustaceans and molluscan shellfish are laid down (refer to Table 9.II, Food Category 2.4.1). 4.6.2 International requirements
International requirements are based upon having a risk-based, preventive management system in place at all stages of the supply chain. The requirements for this preventive approach is described as follows (37): • Fish and fishery products should be prepared in plants certified by the local competent authority. All certified plants should comply with the GHP (39); • The fisheries industry should take responsibility, implement and maintain safety management systems based upon HACCP; • The national competent authority is responsible for the certification of fish processing and manufacturing plants, verification of effective systems, and
issuing of certificates of compliance for export products. This includes auditing and inspection programmes; • National surveillance and monitoring programmes should be in place to demonstrate that all identified hazards are under control, e.g. biotoxins, and to identify potentially emerging hazards; Within this requirement is the flexibility for countries and trading blocks to set specific standards required to deliver an equivalent requirement with those at national level. Such standards need to be science-based, especially if they are more stringent that those recommended by the Codex Alimentarius Commission. Verification that GHP have been followed during the handling and preparation of crustacean shellfish for consumption is usually done by assessing the level of E. coli and/or coagulase-positive staphylococci in the products. Legal requirements generally require that these groups of indicator organisms are present in levels <100 cells/gram of product. Pathogenic bacteria such as Salmonella spp. should be absence is 25 grams multiplied by the number of subsamples tested. 126.96.36.199 United States The FDA brought into force a Federal Mandate Seafood Rule in 1995 which forms the foundation for good practices required for the handling and processing of imported fish and fishery products into the U.S. The best practices include GHP and HACCP (37). All fish and fishery products produced in and imported into the U.S. after 17 December 1997 are required to demonstrate that HACCP principles were applied to their handling and processing. The USFDA has a zero tolerance for Salmonella in fish and fishery products; this includes farmed shrimp. The US FDA considers that the presence of pathogenic microorganisms in seafood is an indicator of adulteration (32). 4.7
Encarta 1997. Encarta 97 Encyclopedia World English Edition. Microsoft. The International Commission for Microbiological Specifications for Food. Fish and Fish Products, in Microorganisms in Foods 6. Microbial Ecology of Food Commodities. Ed. The International Commission for Microbiological Specifications for Food. New York, Kluwer Academic/Plenum Publishers. 2005, 174-249. Cann D.C. Bacteriology of shellfish with reference to international trade, in Handling, Processing and Marketing of Tropical Fish. Ed. Tropical Products Institute. London, Ministry of Overseas Development. 1976, 377-94. D’Aoust J.Y., Gelinas R., Maishment C. Presence of indicator organisms and the recovery of Salmonella in fish and shellfish. Journal of Food Protection, 1980, 43, 679-82.
FISH AND SEAFOOD 5. 6. 7. 8. 9. 10. 11. Fraiser M.B., Koburger J.A. Incidence of salmonellae in clams, oysters, crabs and mullet. Journal of Food Protection, 47, 343-5. Reilly P.J.A., Twiddy D.R., Fuchs R.S. Review of the occurrence of Salmonella in tropical shrimp. FAO Fisheries Circular No. 851, 1992, (FIIU/C851). Reilly P.J.A. Salmonella and Vibrio cholerae in brackishwater cultured tropical prawns. International Journal of Food Microbiology, 1992, 16, 293-301. Twiddy D.R. Antibiotic-resistant human pathogens in integrated fish farms. ASEAN Food Journal, 1995, 10, 22-9. Olsufjew N.G., Petrow V.G. Detection of Erysipelothrix and Listeria in stream water. Zhurnal Mikrobiologii Epidemiologii i Immunobiologii, 1959, 30, 89-94. Anon. First Listeria finding in crabmeat confirmed by FDA. Food Chemistry News, 1987, 29 (14), 38. Ryser E.T., Marth E.H. Incidence and behaviour of Listeria monocytogenes in fish and seafood, in Listeria, Listeriosis and Food Safety. Eds. Ryser E.T., Marth E.H. New York, CRC Press. 2007, 617-54. Anon. Mexican crabmeat, Greek pasta automatically detained. Food Chemistry News, 29 (19), 43. Anon. FDA checking imported domestic shrimp, crabmeat for Listeria. Food Chemistry News, 1987, 29 (24) 15-7. Archer D.L. Review of the latest FDA information on the presence of Listeria in foods. WHO Working Group on Foodborne Listeriosis, Geneva, Switzerland. 1988, 15-9. Anon. FDA to sample shrimp for Salmonella, Listeria. Food Chemistry News, 1988, 30 (36), 9-10. Anon. Monitoring of changes to listeriosis problem urged. Food Chemistry News, 1989, 31 (20), 13-7. Richmond M. Report of the Committee on the Microbiological Safety of Food. London, HMSO. 1990, 133-7. Swartzentruber A., Schwab A.H., Duran A.P., Wentz B.A., Read R.B. Jr. Microbiological quality of frozen shrimp and lobster tail in the retail market. Journal of Food Protection, 1980, 40, 765-9. Wentz B.A., Duran A.P., Swaartzentruber A., Schwab A.H., McClure F.D., Archer D., Read R.B. Jr. Microbiological quality of crabmeat during processing. Journal of Food Protection, 1985, 48, 44-9. Garren D.M., Harrison M.A., Huang Y-W. Clostridium botulinum type E outgrowth and toxin production in vacuum-skin packaged shrimp. Food Microbiology, 1994, 11, 467-72. Molitoris E., Joseph S.E., Krichevsky M.I., Sindhuhardja W., Colwell R.R. Characterisation and distribution of Vibrio alginolyticus and Vibrio parahaemolyticus isolated from Indonesia. Applied and Environmental Microbiology, 1985, 50, 1388-94.
12. 13. 14.
15. 16. 17. 18.
CRUSTACEAN SHELLFISH 22. Varma P.R.G., Iver T.S.G., Joseph M.A., Zacharia S. Studies on the incidence of Vibrio cholera in fishery products. Journal of Food Science and Technology, 1989, 26, 341-2. Madden J.M., McCardell B.A., Morris J.G. Jr. Vibrio cholerae, in Foodborne Bacterial Pathogens. Ed. Doyle M.P. New York, Marcel Dekker. 1989, 525-42. Twedt R.M. Vibrio parahaemolyticus, in Foodborne Bacterial Pathogens. Ed. Doyle M.P. New York, Marcel Dekker. 1989, 543-68. Oliver J.D. Vibrio vulnificus, in Foodborne Bacterial Pathogens. Ed. Doyle M.P. New York, Marcel Dekker. 1989, 569-600. Pearson A.M., Dutson T.R. Quality Attributes and Their Measurement in Meat, Poultry and Fish Products. Advances in Meat Research Series. Volume 9. London, Chapman & Hall. 1994. Reinhard R.G., McAdam T.J., Flick G.J., Croonenberghs R.E., Wittman R.F., Diallo A.A., Fernandes C. Analysis of Campylobacter jejuni, Campylobacter coli, Salmonella, Klebsiella pneumoniae and Escherichia coli 0157:H7 in fresh handpicked blue crab (Callinectes sapidus) meat. Journal of Food Protection, 1996, 59 (8), 803-7. Buchanan R.L., Palumbo S.A. Aeromonas hydrophila and Aeromonas sobria as potential food poisoning species: a review. Journal of Food Safety, 1985, 7, 15-29. Palumbo S.A., Maxino F. Williams A.C., Buchanan R.L., Thayer D.W. Starch ampicillin agar for the quantitive detection of Aeromonas hydrophila in foods. Applied and Environmental Microbiology, 1985, 50, 1027-30. Wekell M.M., Manger R., Colburn K., Adams A., Hill W. Microbiological quality of seafoods: viruses, bacteria and parasites, in Seafoods: Chemistry, Processing Technology and Quality. Eds. Shahidi F., Botta J.R. Glasgow, Blackie Academic and Professional. 1994, 196-219. Schantz J., McKerrow J.H. The dangers of eating raw fish. New England Journal of Medicine, 1989, 320, 1143-5. Ahmed F.E. Seafood Safety. Seafood Safety. Committee on Evaluation of the Safety of Fisheries Products, Food and Nutrition Board, Institute of Medicine. Washington D.C., National Academy Press. 1991. Blackwell J.H., Cliver D.O., Callis J.J., Heidelbaugh N.D., Larkin E.P., McKercher P.D., Tayer D.W. Foodborne viruses: their importance and need for research. Journal of Food Protection, 1985, 48, 717-23. Kapikian A.Z., Chanock R.M. Norwalk group of viruses, in Virology. Eds. Fields D.M., Knipe D.M. New York, Raven Press. 1990, 359-77. Horner R.A., Postel J.R. Toxic diatoms in western Eashington waters (US west coast). Hydrobiologia, 1993, 269/270, 197-205. Perl T.M., Bedard M.S.M., Kosatsky T., Hockin J.C., Todd E.C.D., Remis R.S. An outbreak of toxic encephalopathy caused by eating mussels contaminated with domoic acid. The New England Medical Journal, 1990, 322, 1775-80.
23. 24. 25. 26.
34. 35. 36.
FISH AND SEAFOOD 37. Ababouch L., Gandini G., Ryder J. Causes in Detentions and Rejections in International Fish Trade. FAO Fisheries Technical Paper 473. Rome, Food and Agriculture Organisation of the United Nations. 2005. Australian Government. Australian Quarantine and Inspection Service Hazard Analysis and Critical Control Point (HACCP). A Guide to Compliance with Export Control (Fish and Fish Products) Orders 2005. FAO/WHO. General Hygiene Requirements. Rome, Food and Agriculture Organisation of the United Nations. 2003.
Further Reading Spoilage and pathogens
Gram L., Huss H.H. Fresh and Processed Fish and Shellfish, in Microbiological Safety and Quality of Food Volume 1. Eds. Lund B.M., Baird-Parker A.C., Gould G.W. Maryland, Aspen Publishers Inc. 2000, 472-506. Ababouch L., Gandini G., Ryder J. Causes of Detentions and Rejections in International Fish Trade. FAO Fisheries Technical Paper 473. Rome, Food and Agriculture Organisation of the United Nations. 2005. International Commission on Microbiological Specifications for Foods. Fish and fish products, in Microorganisms in Foods 6. Microbial Ecology of Food Commodities. Ed. International Commission on Microbiological Specifications for Foods. London, Blackie Academic and Professional Press. 1998, 130-89. Sutcliffe P., Disney J. Handling, Processing and Marketing of Tropical Fish. London, Tropical Products Institute. 1977. Connell J.J. Control of Fish Quality. Oxford, Fishing News Books. 1995 Ward D.R., Hackney C. Microbiology of Marine Food Products. New York, Van Nostrand Reinhold AVI. 1991 Pearson A.M., Dutson T.R. Quality Attributes and their Measurement in Meat, Poultry and Fish products. Advances in Meat Research Series. Volume 9. London, Chapman & Hall. 1994.
4.8.2 Pathogens - general
International Commission on Microbiological Specifications for Foods. Microorganisms in Foods 5. Characteristics of Microbial Pathogens. Eds. International Commission on Microbiological Specifications for Foods. London, Blackie Academic and Professional Press. 1996. Varnam A.H, Evans M.G. Foodborne Pathogens - An Illustrated Text. London, Wolfe Publishing Ltd. 1991. Doyle M.P. Foodborne Bacterial Pathogens. New York, Marcel Dekker. 1989.
and before significant spoilage takes place. SMOKED AND DRIED FISH Dr. split and opened out flat. to assure a salt concentration in the fish flesh that is appropriate for flavouring purposes prior to the use of other preservation techniques. It includes the salting. rather than for preservation itself. alternating fish layers with a layer of salt. fish is immersed for a few hours in a relatively low concentration brine. both to preserve and to create a variety of new products. and marinating processes.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. and the extracted moisture is allowed to drain away. and is sometimes known as ‘pickling’. where the salt replaces moisture in the fish flesh. The fish is headed. and a preliminary operation to some smoking.5. The fish is immersed in a brine solution in which different salt concentrations can be used according to the process. 93 . Salting is both a method of preserving fish. dependent on the country or the type of product. The immersion in concentrated brines for long periods is usually used for fatty species and for longer-term preservation. and salting is done by stacking the fish in layers. their combinations and/or variations. CURED. Wet salting or brining. drying and smoking processes. Usually. This can be called ‘kench’ curing and is mainly used for non-fatty white fish. The processes take anywhere from a few hours to a few days or even months. Simon Derrick The Grimsby Institute Humber Seafood Institute Europarc Grimsby DN37 9TZ United Kingdom 5. Fish curing is practised worldwide. drying. Fish can be salted in three different ways or their combination: Dry salting.
spices). The product is dried by contact with heated surfaces or by radiation. depending on the type of product. where an organic acid. thus delaying spoilage for a few days. Fish in contact with refrigerated plates freeze. Usually. Vacuum drying. The fish is mixed with dry salt but in a watertight container. usually vinegar. After the freezedrying process. Traditionally. Food products can be dried in three different ways: Air or contact drying. this ‘brine’ can be collected and placed again over the fish several times. Included in this group are some lightly preserved products where no acid is added to the marinade. As the water is drawn out of the flesh by the salt a brine is formed. There are some marinating processes where salt is used for the first cure and a firmer product is obtained. It is not commonly used for fish products. Marinating or pickling. 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. Drying is the preservation of fish by removing the moisture content of the fish flesh by evaporation. The products are usually consumed raw. and the air movement removes the vapour. it can also be named ‘pickling’ or ‘blood pickling’. thus making the product unsuitable for microbial growth. the product rehydrates quickly but does not retain reabsorbed moisture on cooking. as concentrated brines are formed. without other preservation methods applied after boiling. evaporated water is removed by vacuum pump. Salt. such as ‘gravad’ fish. Heat is transferred to fish from heated air or a heated surface. This is the common method of drying fish. Marinades are acidified brines. Freeze drying. Smoking is an ancient process for preserving and flavouring food. prior to or during the boiling process. thus reducing the water activity of the product. The principle of preservation of the first method is the inactivation of enzymes and killing the non-spore-forming organisms.FISH AND SEAFOOD Dry / wet salting. under normal pressure. the ice sublimes and the vapour is removed from the fish by vacuum pumps. and the fish can be immersed in such brines for long periods. this was done naturally by exposure of the fish to the air. is used as a preservative instead of salt. Salted-boiled fish are the products obtained on either boiling fish in a brine. Salt-boiling. or boiling it with dry salt and water. Longer shelf life is achieved in the products when salt is added. It has found limited application with fish. as well as other flavourings (sugar. This is a mixture of a dry and a wet salting process. the preservative effect is due to the low pH used in the process. are added to the marinade. When the process is running in a closed system.
With the exception of some Asian countries. Cold-smoking. which impart the flavour of smoke to fish. In this process. After the salting step. 95 . Smoking can be either traditional.2 Initial Microflora The initial microflora are those of the raw fish from which the products are made. Hot-smoking. the fish is subjected to heat for a sufficient period of time to coagulate protein throughout the fish. In this process. The different spices used in the immersion brines are also a potential source of microorganisms. temperatures below 30 °C are used. or liquid smoke. by exposure to the wood or sawdust smoke. smoking is now used mainly to give the fish a characteristic flavour. which irreversibly combine with the haemoglobin in the tissue to maintain the red colour. It can be injected into the product. 5. by exposure to the wood or sawdust smoke. and filtered to remove tars and particulate matter. the fish is subjected to heat for a period that does not coagulate protein. process temperatures are over 60 °C. including pathogens. Smoked fish is prepared by subjecting the fish to the direct action of smoke from wood or wood sawdust burning. the sugar used in some products may contain yeasts and moulds as well as spore-forming bacteria. Usually. or added to the brine with other flavourings. and the preservative effect is only slight. or by means of smoke-flavouring processes with liquid smoke. the salt used in the processes may contain halophilic bacteria. 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. leaving only elevated levels of carbon monoxide. Clear smoke. The secondary objective is to impart sensory characteristics such as smoke colour and smoke flavour to the product. Liquid smoke is a natural aqueous condensate of wood smoke that has been aged. It can be smoked either traditionally.CURED. Smoked-flavoured fish. or by means of smoke flavouring processes. However. sporeforming bacteria or osmophilic moulds. such as injection or dipping in a solution of liquid smoke. Also. It is commonly used in place of traditional smoke. fish can be smoked by means other than the direct action of smoke. SMOKED AND DRIED FISH of smoke. The process was designed so that the smoke is filtered to remove all smoke particles. The smoke itself may contain spores of moulds. A dry or wet salting step is always used in the process before exposure to the wood or sawdust smoke. Usually.
8 211.3 Processing and its Effects on the Microflora Owing to the wide variety of products. When a 100º brine is used.I). etc.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. processes and producers’ private recipes. Therefore. it is important to understand the concept of salt concentration. it causes swelling because the fish absorbs water from the salt solution (1). Different brine strengths are required according to the curing processes and/or the type of product. about 50º (132.6 g of salt or more per litre of brine) may cause salt burn in this way. Salt in the water phase is the amount of salt compared with the total amount of water and salt in the fish.6 132. a 0º brine saturation means pure water. is employed.2 237. which is the percentage of salt in the whole weight of fish that is water plus protein plus fat.FISH AND SEAFOOD 5.80º brine (184.4% salt (Table 5. This should be distinguished from the salt content. For smoking salmon. When working with salting. A 100º brine solution means that the solution is saturated and no more salt can be dissolved. only the most common processes can be described here.2 105.4 52.6 264. The dissolution of salt in water is dependent on the temperature.4 184. Thus. a 90º or above brine (237. or salt in the water phase.8 79. % 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. TABLE 5.0 158.8 211.0 . When a milder brine. the fish is usually salted with a 70 . and salt content. smoking or marinating processes. At 18 °C a 100º brine is produced when the solution contains 26. The strength of the solution is described as degrees of saturation (brineometer degrees). Salt in the water phase links the two conditions of moisture and saltiness. Salt concentration means the percentage of salt in the water contained in the fish. it is important to state the temperature. all using salting steps. sometimes the salt crystallises onto the surface of the fish. it is impossible to establish standardised processing stages in the curing of fish.2 g of salt per litre of brine).0 g of salt per litre of brine).
It is common for the dried salted cod in the Mediterranean countries to be made using solar salts. which consequently inhibits the diffusion of the water from the bulk of the product to the surface. However. for which there is a high demand in Mediterranean countries.3. See Figure 5. and salt can speed this oxidation process. can lead to the decomposition of the centre of thick muscles before it reaches the level of salt concentration sufficient to prevent spoilage. When pure salt is used.3. if very pure sodium chloride is used for curing purposes. This is because fatty fish turn rancid quickly. ‘Salt burn’ might occur when the surface of the fish dries out too quickly as a result of the small grain size of the salt. The most common fish used in the northern hemisphere is cod (Gadus morhua). but the concentration of copper and iron must be kept under 0. and for a heavy salting a ratio of 1:3 salt:fish is used (1). SMOKED AND DRIED FISH 5.15% calcium and magnesium. are likely to cause hydroscopicity of the product and. 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. It takes about 3 weeks until proper salt penetration and moisture loss are achieved. 97 . outgrowth of halophilic bacteria may occur.15 . the fish is mixed with one part of salt per eight parts fish.1. having a short section at the end of the tail. addition of levels of 0. This condition is known as ‘putty fish’ (4). mainly in tropical conditions. however. For a light salting. The calcium and magnesium impurities. Usually it is followed by other preservation techniques such as drying and/or smoking. and essentially free from iron and copper (3). and a ‘putty’ fish product is also obtained (1).1. 5. when the air humidity is high. lean white fish is used for this purpose. On the other hand. and the salt has penetrated throughout the fish (3). Salting can be done in a number of different ways. Also. it tends to lead to a slight yellowing and softening of the product. producing characteristic pink patches. which include calcium and magnesium. and such delays.1 ppm and 10 ppm. caught in North Atlantic waters.CURED. when rock salt is used the ‘dun’ patches caused by moulds are prevalent (1). respectively. are recommended to help make the product whiter.1 Salting Salting of fish for preservation purposes has been used since pre-history. the fish is restacked with fresh salt after about a week. can be dried further to produce ‘klipfish’ or ‘bacalão’.1 Dry salting / Kench curing Usually. low in calcium and magnesium.0. The fish is then stacked in piles of about 2 metres and the water is allowed to drain away. The salt used should be of food-grade quality. A ‘green cured’ product is obtained when the water content has been reduced to about two-thirds of the initial amount. ‘Green cured’ split cod with about 60% of the backbone removed.35% and 0. respectively (1).
Food-poisoning organisms. Where fish are salted in their whole form.1.90 are rarely achieved. The greatest barriers to salt penetration are the skin and scales. when a light brine is used.5. Processing of dried salted cod 5. eviscerated and/or split). it appears that water activities much less than 0. such as 98 . so that the risk of spoilage is high. and the uptake of salt is dependent on the type of fish (fatty or lean) as well as its presentation (whole. For large fish it is advisable to split it before salting. the process is mainly for flavouring purposes prior to the use of other preservation processes.3.2 Wet salting or brining As referred to above. Fig. Only small fish can be salted whole. such as Staphylococcus aureus could survive and grow at such water activities in the product. although toxin production would be unlikely (4).1.FISH AND SEAFOOD Dry salting can also be used for small species without the need for opening the fish flat.
they can be stored at 28 °C.23 3.64 4.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. sardines.95 3.5 4. For preservation purposes.56 6.11 9.76 4. the brine is then replaced over the fish.91 2.3 Dry / wet salting In this process.51 3.84 1.44 2.23 8. The main objective of the brining method of salting fish is to limit air penetration during processing.78) (1).17 4. the fish is dry-salted. and can be collected and replaced again over the fish.23 2.05 7.3 days.26 4.03 4.CURED. limiting the amount of rancidity that can develop.76 4.04 5.26 7. the water that comes out of the fish forms a brine in which the fish stays immersed for 2 .19 3.25 5. the fish enzymes play an important role by maturing or ripening the product.03 2.74 9.1. It is then taken out and dried in the sun or in kilns until about 35% moisture in the product is obtained.30 2. SMOKED AND DRIED FISH smoking.0 3.85 3.41 5.98 6.78 2.09 8.II.26 3.86 2.49 4.51 5.65 2. Histamine formation is of importance when using scombroid and some clupeidean species.66 5.99 2. with a shelf life of about 4 weeks (water activity (aw) about 0.5 10.70 3. Histamineforming bacteria do not generally grow at water activities below 0.76 4.70 6.41 4.98 6. herrings and other fatty fish can be used.66 5.47 6. thus. Fresh sardines. See Table 5.3. but mackerel. can be cured in a saturated brine solution for 6 days and.22 6. a saturated brine is used. if split fish is dry-salted.45 3.54 5.5 3. and a red burgundy colour will develop in the product. headed and gutted.44 2.33 5.46 4. the product obtained is called gaspé-cured (5).23 2. with pressure being applied to the top of the container to help the brine to drain away.90 (1).78 6. The fish are kept immersed in their own brine near saturation point for several days. after being drained and pressed at 465 kg/m2 for 15 hours.09 8.88 5.5 3. Here.16 7.17 5. Anchovies are the main fish used for these purposes.26 2.26 1.41 6.61 3. For example. This is why it is the preferred method for fatty fish.0 2. 5.98 5.80 5.06 4.0 4. The brining step for smoking purposes also helps the product to develop a desirable glossy appearance.60 2.76 4.25 5.85 3. the liquid that comes out of the fish forms a brine.96 1.11 4. There are several variations of this process.35 4.10 1.76 4.76 99 .73 1. TABLE 5.11 3.00 3. but as it is placed in containers.41 3.61 3.
because of the salt content of the product. they can be cut and cured for 3 days in a solution with 3% white distilled vinegar. and spices) is added to the brine.2 months. Another salt-boiled Indonesian product. usually requiring refrigeration below 10 °C for longer-term storage. underlines the need for careful control (4). even if the product is hermetically sealed. Usually. They will have a salt content >6% (w/w) in the water phase. or a pH <5 and they are stabilised for a limited period. A much firmer product is obtained when the fish is salted prior to acidification. herring or other fatty species are used. All these products are ‘semi-preserves’. or can be boiled by being placed in a basket. outgrowth of bacterial spores after processing may occur (4). salt. ‘ikan kaju’. only the nonspore-forming organisms are eliminated in the process and.3. placed in a marinade of lime juice. and lasts for 3 .3. The Indonesian ‘pindang’ is made by boiling the fish in a saturated salt solution and it is sold as is. until the flesh is soft. but it has further smoking. After this.7%) for as long as 3 weeks. and 6% salt. giving a cooked appearance to the product. There is epidemiological evidence that these products have been the cause of some 100 .2% acetic acid and 2 . then packed in barrels with a 70º brine. Afterwards. Ceviche. and packed (3). 5. Another variation uses sour cream instead of the second brine step. whereas the latter method preserves the fish for only a few days.4% of salt) for distribution and sale (1). In a second step. a Peruvian product. flavouring (sugar. the occasional implication of pindang in cases of foodborne illness. Traditional jellied eels are a product where pieces of fish are cooked in boiling water. they are packed for consumption within 2 weeks at chill temperature (6).5% 120grain distilled vinegar. Other preservation techniques can be used for extending the shelf life of these products. is made of different varieties of fish. it can be repacked in milder brine (1 . The first process can give a longer shelf life. The product is usually consumed raw with the high strength acid solution softening the bones enough for them to be eaten. drying and salt-boiling steps until a hard translucent block is obtained (4). up to 1 .3 Marinating / pickling Fish can be brined in solutions based on organic acid instead of salt.30 minutes. As referred to above.6 days without refrigeration (1). containing 1 kg of salt for every 25 kg of fish. The fish can be soaked in a high concentration of vinegar (about 4 .FISH AND SEAFOOD 5. 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 . Once cooled. However.2 Salt-boiling Fish can be boiled after being dry-salted in a watertight container and heated for at least 1 hour. The lime juice plays the role of the vinegar. usually flavoured with onions and hot chillies. has a shelf life of several years. The cooked portions are poured into bowls containing gelatin dissolved in water (10% solution) and sometimes a small amount of vinegar. and even death.
5. In the first case. and the stacks are placed in the refrigerator. a very fine delicacy for Oriental food lovers. sea cucumbers and seaweeds. 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). The most common method used for fish products is the air-drying process. some fish species such as moray eel and varieties of dog fish. such as squid. Matjes are made of fat young herring. used as an appetiser. split and dried gadoid species) are traditionally made in cold climates. In Scandinavian countries. Usually. air flows naturally. At pH levels below 4. which are placed in a mixture of salt.) where no acid is added to the process. Fresh dill is placed between the layers of fish. by using either solar heaters or mechanical dryers. bacterial toxins and parasites (7). and a fine product is obtained. etc. are generally air-dried at sea. Stockfish (dressed. Shark pectoral fins. Italy and Spain (5). 101 . Some bacteria and enzymes are still active in the presence of acid and salt. but sometimes it is dried without prior salting. jellyfish. SMOKED AND DRIED FISH foodborne diseases related to the presence of biotoxins. it being preferable to use frozen salmon or to freeze the product at the end of the process to avoid parasite survival.There are several products named marinades (matjes. They are not cooked prior to consumption and they have a limited shelf life even at refrigeration temperatures. In some coastal areas of Portugal. Other products. gravad fish. gutted. sugar and spices. It is important to assure the quality of the raw material. Provided the product is kept in dry and fairly cool conditions.3.CURED.4 Air drying This is probably the oldest method of fish preservation. 5. while in the latter case there is a provision for fans or other means of mechanically moving the air (1). They are an important dried product. such as Listeria monocytogenes and Clostridium botulinum. It takes about 2 days to marinate completely. then mild-cured with special seasonings and packed in barrels with 80 º brine (5). Dried shrimps are widely used in Asia. the fish is dried naturally in a relatively cold environment. it will last for many years (5). are likely to survive in these mild-processed products. are sundried and. the temperature and the amount of pressure applied (5). marinated products will keep for several months at a temperature of 4 °C.0) (7). under a light pressure. South America and the Caribbean. Gravad fish is a Scandinavian product made of salmon fillets. depending on the thickness. still made without prior salting on a large scale. Stockfish products have a golden brown colour and are very hard. and the flesh will break down. the fish is dried after salting or brining. Stockfish is extensively used in Portugal. are commonly dried in Asian countries. Foodborne pathogens. mixed with salt. 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. Dried octopus is also a very common dish in the interior of the country. after frying.
making a water barrier and consequently. but fast enough to avoid spoilage caused by bacterial and enzymic activities (1). typically in stews and soups. the quality is more difficult to control. Dried salted products must have a water activity below 0. Owing to restriction of fishing areas for Spanish and Portuguese vessels since the 1970s. Although controls are in place to ensure that products processed in such relatively uncontrolled conditions do not enter the supply chain.III. They must be soaked in water. more commonly nowadays. In many such artisanal fisheries the drying is simply conducted by spreading the fish on the beach. the skin will form a hard case (case-hardening). aureus) or spores (e. or wooden drying rack to dry in the sun. which leads to mould growth (aflatoxins. skin up. and has the primary objective of creating a product with specific taste and textural characteristics. To prevent this. C. some of which are listed in Table 5. In conclusion. by warm air circulating in a drying chamber. tarpaulin. The fish must be stored in reasonably dry conditions to avoid moisture absorption. the protein must be set or denatured with low-temperature drying before applying higher temperatures. Portuguese and Spanish fisheries cannot satisfy their own countries’ demand for cod. and increasing amounts of such products are making their way into the market place. dried salted cod (bacalão) is now mainly produced in Norway. Thus. and the water must be changed several times before cooking.g.g. the product can be held for months at room temperature. Staph. botulinum). 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 diffusion of the bulk water to the surface of the fish is blocked. Iceland and Canada. the better the moisture transfer. Italy and South America. The amount of moisture transfer from the fish to the air is dependent on the relative humidity . Spain.83 (9)).the lower the humidity. When no more water can be removed from the fish. France. neither it. there is an increasing demand from people that have emigrated to industrialised nations for traditional products. if the rate of water removal is too high in comparison with the diffusion rate. After dry salting.75 (1). Whilst the low aw of the final product will inhibit microbial growth. spoilage and possibly C. the fish must be dried slowly enough to avoid case-hardening. A highly demanded dried fish product in Mediterranean countries is dried salted cod. Whilst the production of dried fish products is tightly controlled in industrialised countries. toxic secondary metabolites of the moulds Aspergillus flavus and Aspergillus parasiticus are not produced at aw <0. However. and it is exported to Portugal. botulinum growth might occur in the bulk of the product.FISH AND SEAFOOD When the fish is sun-dried in tropical conditions. 102 . nor subsequent cooking operations will affect the presence of toxins (e. that may have contaminated the product during the drying process. the ‘green cured’ cod is rinsed or washed with water and then hung to dry either by the sun and breeze or. Traditional processing conditions provide ample opportunity for product contamination from a wide range of hazards and sources. The product is dried to a final water content of about 25 to 38% (3). It is important to balance the drying rates and quality/safety issues.
salmon. eel. Staph. 5. The smoking procedures vary widely. aureus (toxin) C. However. clams. Blends of various woods can be formulated to produce sawdust with specific flavours. using electrostatic filtration of the smoke. temperature and humidity can be automatically controlled and the process is faster because the directed air movement draws the smoke through the fish. Aroma and flavour are a blend of smoke components (3). It has been reported that the smoke from mahogany.5 Smoking Smoking today. as referred to above. Preventing the occurrence of carcinogens during smoking has been an issue for the past several years. SMOKED AND DRIED FISH TABLE 5. mussels and many others from tropical countries. tuna.CURED. haddock. Hardwoods are considered better because of the milder flavour they impart to the food. making full description impossible here. such as Escherichia coli. cod roes. white mangrove and abura has an inhibitory effect on some microorganisms. are some of the ways of reducing the polycyclic aromatic hydrocarbon (PAH) compounds (10). is mainly a process for flavouring fish rather than a preservation technique.3. sprats. oysters. It can be used for cold or hot smoking. 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. The production of smoke by wood or sawdust burning can be done in a mechanical kiln or in traditional chimney kilns. botulinum (spores) Pesticides / Chemicals Physical Glass / sand etc. swordfish. aureus and Saccharomyces cerevisiae (10). softwood colours the product quickly. trout. Proper cleaning and preparation prior to smoking and washing of the fresh or frozen fish by water spray or a continuous 103 . Top-quality fish must always be used. There are many fish species that are used for smoking purposes . herring. cod. and the use of smoke generated by superheated steam or liquid smoke. The quality of the smoke is related to the type of wood used. mackerel. In the mechanical kiln.
but this concentration will again depend on the type of product and the length of the brining stage. temperatures must be kept under 30 °C throughout the process. or packed in retortable foil pouches. but it will increase towards the end of the process (10). it will take up salt twice as fast as fresh fish (10). Thus. such as smoked salmon-trout. but a ‘safety’ (i. Drying should be minimal at the start. 70 – 80 º brine is commonly used.18 hours at 4 °C.3. skin on or off).5. Also. The salting period is dependent on the size of the fish and its presentation (whole.FISH AND SEAFOOD water flow system with chlorinated water (25 . are not cooked prior to consumption. temperatures and times used in processing coldsmoked fish are very favourable for the proliferation of food-spoilage and foodpoisoning microorganisms. after being smoked need to be chilled immediately to 4 °C before being packed. it will reduce the salty flavour. tuna and swordfish.10% of the weight of the fish. owing to the elution of soluble proteins. This difference is crucial for safety issues because the flesh is not cooked. Sugar can be added together with the salt. and the cold smoke is denser. However. giving the product the desired appearance and flavour (4). skin off or on. as it retains more moisture. all cold-smoked products should be cooked before consumption. which is important for the appearance of these products (4). thus allowing the deposit of smoke on the fish surface. Depending on the type of product. spices. usually dry-salted and cold-smoked. Wet salting or brining is preferable to dry-salting when smoking fillets because. Some herring products. 104 . canned or bottled. whether cold.e. All the smoked products. In cold-smoking. When frozen/thawed fish is used. The fish can be dry-salted using a weight of salt 5 . covering the fish surface completely (10). an attractive glossy pellicle on the fish fillet surface is achieved after draining. the following step is salting. pasteurisation) temperature is not achieved in any stage of the process. permitted colourings or smoke flavours can also be added to the product in the brining step. 5. smoked cod fillets and finnan haddock are usually cooked before consumption. as well as many other products prepared in the same way. The maintenance of temperatures below 30 °C avoids protein coagulation.1 Cold-smoking The major difference between cold. it will double the drying time in the kiln to get the same total weight loss as for dry-salted fillets (11). filleted. If wet salting is chosen. If frozen fish is used. Cod roes. wooden boxes or master cartons (10). gutted or filleted. Thus. The length of the smoking process for such products is much greater than for the hotsmoked fish. Brining at 10 15 °C is recommended (10). the highly desirable cold-smoked salmon. are boiled before smoking (12). Sugar. they can be vacuum packaged. shrink packaged.or hot-smoked. however.50 ppm) should always be done just before process initiation (3). Whether the product is prepared headed. low temperatures will slow the uptake of salt. the excess salt is removed with water. or quick-frozen and stored at -18 °C (2).and hot-smoking is the temperature of the process. thawing must be done slowly for 12 .
5. botulinum type E the product must have at least 3% salt in the water phase and be stored below 5 °C. if a 105 . The protein must be set or denaturated with lower temperatures before the application of higher temperatures to avoid case-hardening. Although other human pathogens that can grow at refrigeration temperature. typically mesophilic. can grow with salt levels of 10% in the water phase. Pathogenic vibrios. there is no point in the process that can fully assure the absence of L.2. and hotsmoked salmon (kippered) (3). a partial cooking period at 50 °C. If the refrigeration chain is maintained.2 Hot-smoking Hot-smoking must use temperatures between 70 and 80 °C at any stage of the process to ensure protein coagulation throughout the product. Vacuum packing extends the shelf life of the lightly smoked products by suppressing the growth of other bacteria and moulds. but the proteolytic types A and B. a final cooking period at 80 °C (12). This temperature should be maintained during storage and distribution (3). condensation will occur inside the pack. thirdly. refrigerated storage and vacuum packaging still allow its growth (12).CURED.3. in order to produce toxin.5. Hot-smoked products require a longer cooling stage. On the other hand. Vacuum packing is recommended only for coldsmoked salmon. stored at 4 °C and packed. Smoked products can be also packed under controlled-atmosphere packaging. See Figure 5. Length of the smoking process at each stage will depend on the species and the type of product. a drying period at 30 °C. monocytogenes. thus allowing any psychrotrophic C. and the recommended gas mixtures are 60% carbon dioxide and 40% nitrogen (15). botulinum type E present an even longer time than before to grow and produce toxin (14). Furthermore. which underlines the danger of storage at abuse temperatures of >10 °C (13). if present. Smoked salmon fillets are usually vacuum packed whole or after slicing. To avoid growth of non-proteolytic C. an initial humidity of 70% seems to be preferable because it is a good compromise between maximum smoke adsorption and minimum casehardening (1). are of concern when smoking fish from tropical waters (13). the products are cooled. gravad salmon (also called gravad lax or gravlax). 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. secondly. Afterwards. but they should be cooled to a temperature of 10 °C within 3 hours. During smoking. The exception is vacuum packed fish. can be isolated from seafood. they are more sensitive to sodium chloride and unlikely to grow in smoked products (13). SMOKED AND DRIED FISH During cold-smoking. A typical temperature set is as follows: firstly. and their sensitivity to salt (13). as. These products should be vacuum packed at room temperature. such as Yersinia enterocolitica. if they are taken from a chill room at 3 °C and packed. 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.
5.2. it will have a reduced shelf life. and moulds will grow on the product (12). Fig.FISH AND SEAFOOD product is packed too warm without being cooled to room temperature. Process of cold-smoked sliced salmon 106 .
are used. Aspergillus tamarii) and several Penicillium spp. The classical liquid smoke is 107 . 21. synthetic smoke flavours. to have no further advantage over other packing methods for other fish (12). including ‘umami’ (16. The composition of commercial liquid smoke can vary widely depending on the method of manufacture. a very expensive and fashionable smoked product in the Netherlands. A type of dried hot smoked tuna called masmeen or masmin is produced in the Lakshadweep islands of the Indian Ocean. but mechanical drying is used instead of sun drying. (Penicillum cyclopium and Penicillium puterellii) (19). Katsuobushi. break down of fats and the moderation of smoke flavour. moulding it into long thick sections. such as yeast derivatives. rewetted and remoulded to give the desired shape and consistency. Aspergillus oryzae. Eels. hot-smoked fish can be classified as a ‘heat-treated or pasteurised’ product (7). is a boiled smoked fish product that is used in Japanese cooking to impart flavour.3 Liquid smoke Smoke-flavoured fish represents a small percentage of the total smoked fish industry output. 18). 22. apart from the improved appearance. cooking in brine. Growth of these moulds on the tuna results in the extraction of moisture. including Eurotium repens and Eurotium rubrum. white fish about 8 days. resulting in a characteristic flavour (20. Boiled or steamed clams. Aspergillus ostianus.5. They are generally not cooked prior to consumption. In terms of safety. The production method is similar to masmeen. The product is removed from the drying cycle. 5. Hon kare bushi is a form of Katsuobushi from which shavings are cut to make aromatic stocks for use in Japanese dishes (24). Vacuum packaging retards the onset of rancidity when fatty fish is used. 23). 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). fatty fish will remain in good condition for 6 days. Natural smoke extracts. Mackerel. and many closely related Aspergilli (Aspergillus ochraceus. are usually 80º brined for 5 .6 hours and smoked for 3 . SMOKED AND DRIED FISH There are several species that are usually hot-smoked and the process can vary markedly. 17. are usually hot-smoked after being immersed in 80º brine or dry-salted (6). and substances unconnected with smoke but with a smoky flavour or smell. then hot smoking and finally sun drying it. Masmeen is usually prepared from bonito tuna by eviscerating and filleting the fish. oysters and mussels are usually wet-salted in 50º brine for 5 minutes and hot-smoked at 80 °C for less than 1 hour (12). The product is often allowed to go mouldy by incubating it in a chamber containing a natural fungal flora.3. When refrigerated at 3 °C. chilled or freeze-thawed.CURED. they can be stored for 6 months at -30 °C and for even longer when frozen in vacuum packs (12). Hot-smoked products made from white fish will usually keep better than those from fatty fish. They will keep well frozen. similar to masmeen.3½ hours using the typical temperature set for hot-smoking referred to above (12). but seems.
applying it as a dip after brining. 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). 5.8% and still provide protection for 7 days (25). The European Scientific Commitee for Food. According to some recent research. spraying is preferable because it is easiest to control flavour and acceptable with this method (3). There are two groups of chemicals of concern in smoke: polycyclic aromatic hydrocarbons (PAHs) and nitrosamines. Liquid smoke was considered to provide advantages of safety. the salt level could effectively be reduced from 4. so as to impart an unnatural red colouration to the flesh. it has been reported that the liquid smoke-treated product has less pronounced aroma and flavour (25). drying.03 μg/kg of benzopyrene. Whilst neither the CO or the carboxymyoglobin complex are harmful in themselves.06 μg/kg of benzoanthracene in the final foodstuff (28). pickling and marinating fish were originally used to lengthen the short shelf life of fresh fish. injecting it into the fish flesh. In experiments comparing traditionally smoked fish with liquid smoke.3. It has been shown that liquid smoke can reduce the formation of PAHs (3). or applying it as an atomised spray within a modern automatic smoke kiln. nutritional neutrality and antioxidant activity (27). 5. When liquid smoke was used. or smoke extracts in organic solvents. Smoke-flavoured fish processed using liquid smoke can be prepared by including liquid smoke in the brine.FISH AND SEAFOOD smoke condensate that is dissolved in water or oil. and less than 0. botulinum types A and E spores in several fish species that were stored at 25 °C for 7 and 14 days (25). Fresh fish spoilage may be measured by 108 .4 Spoilage Brining. 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.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. consistency of flavour and colour. both of which are considered potential carcinogens. the result of this process is seen as a manipulation of the natural degradation of tuna colour that hides the true nature of the product quality and therefore has been made illegal in the EU. The process relies on the generation of the smoke to produce elevated levels of carbon monoxide (CO) which interact with haemoglobin in the muscle to form the very stable bright pink/red carboxymyoglobin complex. smoking.6 to 2.5.
Spoilage needs to be carefully defined for each product. chemical oxidation. marination or drying). Endogenous marine fish bacteria prefer to grow in the presence of salt (marine concentration approximately 2. Micrococcus spp. for others. The microflora attached to the fish is relatively unaffected by light brines used before smoking. Pickling and/or the anchovy process uses solid salt extraction of fish fluids to form a brine solution (cure). Within the limits of safety. these bacteria tend to continue to dominate the flora in the liquid. spoilage is due to endogenous microorganisms and enzymes of the fish. When acid marinating or smoking. which are found in the salt. Even saturated brines allow some halophilic organisms to grow. Brining. the fish are cut and splayed to allow greater surface contact with the salt. In salts. mainly comprising corynebacteria and Gram-negative bacteria from the fish and water (29). the pickle and dry-salting preservation processes require further processing for significant shelf life increases (e. a process similar in microbiological effect to that of saturated brining. a product may become undesirable for consumption as a result of changes in its sensory characteristics. and. as the fish are added. or extended refrigerated storage. For some products. the fish are first cleaned (known as ‘rousing’) by a salt rub to remove any outer slime. Upon salt packing. limits the types of microorganism capable of growing. Often. these halophilic organisms may grow and cause spoilage. Once these changes are defined. otherwise. also. SMOKED AND DRIED FISH grading schemes or quality indices. the bulk of the microflora comprises Bacillus spp.3%). for some. Salts used for brining have been found to carry a microbial load of between 10 and 1000 organisms per gram of wet salt. the shelf life may be estimated and investigation made into the causes of the loss of sensory quality. The use of drying and/or salt to lower the water activity. 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. and of smoke to add chemical preservatives. both of these will affect the spoilage patterns. Stronger and longer brining has an effect like that of pickling. contact with the bacteria 109 .g. and coryneform bacteria. When heavy brining or salting is important for product shelf life. but. or their products require refrigerated storage. Once placed into brines. the production process will modify the microbial flora present on the product. which may also be produced to describe the quality attributes and their loss over time for processed fish. This is not usually the case for smoked fish as the residual salt concentration is not high enough to inhibit all the non-halophiles. and high concentrations are usually required to inhibit their growth fully. a very dry product is required to produce 'shelf-stability'. Most marinated and smoked fish products are first brined or salt-pickled. pickling and dry-salting all introduce bacteria to the fish. the consequent reduction in the pH will help to inhibit spoilage organisms.CURED.. rock and manufactured salts did not usually contain halophilic bacteria (29).000 cfu/ml of brine may build up. solar salts investigated carried the highest load and included halophilic organisms as a result of their production method (open-air evaporation of sea water). however. the fish flora are transferred to the brine mix. therefore. Usually..
‘Pink’ describes a condition associated with surface growth of the pigmented halophilic bacteria Halococcus spp. flavus. Sporendonema sebi)). if not too contaminated. rancidity due to lipolysis. most notably Penicillium citrinum and Penicillium thomii. Insufficient salt penetration into the flesh leaves areas where Gram-negative rod-shaped bacteria and micrococci grow and cause a putrefactive spoilage. Unlike ‘pink’. including hydrogen sulphide (26). (syn Oospora halophila) and Polypaecilum pisce are two white halophilic moulds that are associated with spoilage of salted fish (31. ‘dun’ is not characterised by the production of offensive odours. including A. may also be found in the salt used in production. The bacteria form coloured colonies on the surface of salted fish prior to drying. gaspé). Bacteria from these two genera also grow sufficiently in brine and even salt to cause pink discolouration. and Halobacterium spp. B. like the ‘pink’ organisms. the best way of controlling the organisms is to refrigerate the fish. lactic acid bacteria (LAB) and moulds. ‘pink’ and ‘dun’. Aspergillus sydowii. and still produce a reasonable dried product.g.FISH AND SEAFOOD in the salt is made and the flora will develop accordingly in the blood pickle. (syn. Only halophilic microbes may grow. sometimes covering the whole of the fish with a whitish. The metabolism of the organisms will produce off-odours. as the water activity of saturated brine solutions is low enough to inhibit most bacteria. Therefore. The condition describes the growth of the halophilic moulds (Wallemia sebi. A. rubrum are also common isolates from dried fish. were also isolated. E. or fat breakdown) caused by enzymes rather than by microbial growth. Their growth produces noticeable spoilage of the fish appearance but is also controlled by refrigeration. or after drying if the fish is stored in excessive humidity. 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). powdery growth. yeasts and fungi. Several Penicillium spp. as little if any growth will occur below 5 °C on salted fish (29). niger. spore-forming bacteria. Studies conducted on dried and salted fish from Indonesia (31. 33). This condition is often associated with case-hardening. The bacteria are osmosensitive and will lyse in water. Aspergillus wentii. it was found in about 50% of samples examined. repens and E. The microbiology of heavily salted products is dominated by Gram-positive. W. At the pre-drying stage. together with a range of Aspergilli. 34) have shown that P. yeasts. halotolerant or halophilic micrococci. halophilica was isolated 110 . and Aspergillus penicilloides. Eurotium amstelodami. sebi is considered to be the main spoilage agent of dried fish produced in temperate regions (30). though only the latter two species were considered likely to have grown on the fish (32). pisce was the main fungus isolated. which. although it may occur in other salted fish. spoilage of these pickled and dried fish in refrigerated storage tends to be due to chemical changes (textural changes due to proteolysis. protein breakdown. Basipetospora halophila. 32. A number of spoilage organisms can grow and produce conditions such as ‘sliming’. ‘Sliming’ or ‘putty’ fish is a condition that occurs in poorly or insufficiently salted cod (e. it is possible to wash off the infected parts..
as already stated. and microbial spoilage is more likely.. has been isolated from dried fish (35). Pseudomonadaceae. most marine bacteria are moderately halophilic and are not effectively inhibited by these low salt concentrations. Technical studies demonstrate the ability of many of the xerophiles above to grow at typical water activities seen in dried salted fish (32. able to grow on dried salted fish. Fish processed by cold-smoking. including A. Refrigeration is necessary. Aspergillus clavatus and Aspergillus niger. but. The brining to 3 . but this will not prevent the growth of spoilage bacteria. so the most probable spoilage organisms will be spore-formers or post-smoking contaminants. Smoked fish products tend to have a light brine treatment. Marine vibrios were the dominant flora in smoked salmon that was drysalted. whereas a mixture of Enterobacteriaceae and LAB were prevalent in brineinjected salmon (37). It has been reported that the type of salting (dry.5% sodium chloride in the water phase will inhibit some bacteria. The heat during hot-smoking will kill vegetative microorganisms but not all spores.CURED. with a lower aw limit for growth of 0. In Vietnam. The preservative effect of the smoke will inhibit some spoilage microorganisms. where a high internal temperature is reached. it was. The salt in the water phase must be high enough to inhibit the growth of C. around the water activity of saturated sodium chloride.V gives details of pH ranges and aw minima for the growth of some important spoilage microorganisms and pathogens (38). so the bacteria spoiling these products will be psychrotrophic or psychrophilic. SMOKED AND DRIED FISH infrequently at low levels.or wet-salting) in the cold-smoked salmon process leads to different microflora growth at the onset of spoilage. Vacuum packing limits chemical oxidation. Refrigeration of hot-smoked fish will greatly reduce the number of organisms able to grow and will lengthen the shelf life. Table 5. 36). however. LAB. flavus.747. 33. 111 . botulinum type E.IV. Reports for vacuum packed cold-smoked salmon have identified more than one organism associated with spoilage. which does not create the salty conditions required for spoilage by those halophilic microorganisms. but often colour changes due to chemical oxidation of the smoked product will limit the shelf life. Cold-smoked fish will contain microbial flora representative of that of the raw fish. For more information on spoilage see Table 5. not the growth of spoilage organisms. Vibrionaceae and Enterobacteriaceae may all play some part in shelf life limitation. or any smoking process where temperatures in the interior of the fish do not reach >50 °C for a significant period of time. will have a different residual post-smoking flora from 'hot-smoked' fish. Aspergillus spp.
Histamine producers Halophilic spoilage organisms Spore formers and PPC None Acid-tolerant spoilage organisms Xerophilic moulds Fresh fish spoilage organisms.FISH AND SEAFOOD TABLE 5. sebi and B.variable.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. . Matjes v3 + ++ v - +++ ++ ++ - v +/- ++ ++ ++/++/- Pindang Bacalao + +++ Gaspé cure Anchovie ++ Anchovies ++ - - - ++ ++ Saturated brined fish Salted then boiled Salted. LAB.not used/unchanged. + to +++ strength of preservative effect 4 Post-process contaminants 112 . and Halobacterium spp.IV Spoilage of salted. boiled. Enterobacteria and PPC Spore formers and PPC Variable Salt marinades Salted then boiled Brined then boiled Salted then dried Canned Fish Gravad. halophila Halophilic spoilage organisms Halophilic spoilage organisms. Dun. 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.
under optimal conditions.5% to 5%.0-9. although the combination of the two factors.97 0. but if the fish products are stored at temperatures <10 °C. A number of surveys have shown that fish from temperate waters is mainly contaminated with the psychrotrophic C.5. The concentration required to prevent growth at room temperature can vary from as low as 3. temperature ≤5 °C and salt ≥3% in the water phase is sufficient to prevent any growth of type E. and as botulism is a serious hazard in foods. when present in the raw material. smoked and dried. the fermented products being the most dangerous group (7). It has been shown that.5% of salt in the water phase and great care must be taken to avoid any temperatures >10 °C (13). aureus Yeasts Moulds Lactic acid bacteria pH 5. However.CURED. Before. popular demand for healthy foods has led to a change in the tastes preferred by the public and. botulinum type E. In the United States.5 5. salted and pickled products accounted for 10. 9 and 8 of the outbreaks. the importance of cold-chain integrity must be emphasised. 3% sodium chloride in the water phase is enough to inhibit growth of type E for at least 30 days (39).75 0. The presence of salt in cured products has a great effect on the growth of the bacteria. sodium 113 .5-10. and C. pressure on the traditional methods of curing. respectively.V pH ranges and aw minima for growth of microorganisms (38) Organisms Most Pseudomonadaceae C.5 4.8 1. botulinum.86 0. the non-proteolytic type E strains can grow in up to 5% sodium chloride. botulinum type E and production of toxin.97 0. the products were heavily salted. with this change.8 0. could not grow and produce toxin.1 Pathogens: Growth and Survival Clostridium botulinum In the last few decades. although approximately 10% salt in the water phase is necessary to fully inhibit the mesophilic types A and B strains (14). types A and B are not affected by 3 .5 0-11 3. Examination of 165 outbreaks of botulism caused by fish products showed that smoked. but the concentration of salt in smoked salmon or trout (usually varying from 1 4% salt) is not high enough to prevent growth. Sodium nitrite in conjunction with sodium chloride is known for its ability to inhibit C.5-8.61 Organisms Most Pseudomonadaceae C. Since neither the smoking nor the drying parts of this curing process are particularly severe. SMOKED AND DRIED FISH TABLE 5.5-8.9 0. botulinum type E Staph. botulinum Staph.5 5. aureus Most spoilage bacteria Most spoilage moulds Halophilic bacteria Xerophilic moulds Aw 0.5-10 4. but the presence of the mesophilic types A and B is of concern in fish from warmer waters (13).
if recontamination occurs and storage conditions are favourable. but. they will grow rapidly (7). growth and toxin formation may occur. the organism may contaminate the product. if it contains sodium nitrite ≥100 ppm.5. However. after processing. they do not grow well in the raw material. when the bacterial load of the product is lower. its occurrence in a final product is due to contamination during processing by an infected asymptomatic carrier with poor personal hygiene. Shigella and E.3 Enterobacteriaceae The pathogens Salmonella. As with other organisms. monocytogenes is a ubiquitous organism that has been detected in fresh fish skin. The degree of concern in coldsmoked salmon is low unless temperature abuse >10 °C occurs (13). 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. The human carrier rate can be as high as 60% of healthy individuals with an average of 25 .2 Staphylococcus aureus This is a ubiquitous organism but its main reservoir and habitat is animals and. Thus.5. more importantly. 5. In foodstuffs. has been reported (42). An outbreak of Shigella in an Italian cruise ship. As they are poor competitors.5. The organism is mesophilic with a minimum growth temperature of 7 °C. if the person handling the product is a carrier of Staph. but requires higher temperatures for toxin production (>10 °C).FISH AND SEAFOOD nitrite can be added to the brine but this is not permitted in EU countries because nitrites might lead to formation of the carcinogenic nitrosamines. aureus. It is a halotolerant bacterium able to grow at water activities as low as 0. and the minimum pH for growth is 4.5% salt in the water phase.or cold-smoked fish should have at least 3.0 .4 Listeria monocytogenes The psychrotrophic L. if storage temperatures are over 10 °C. the organism can survive most of the light curing processes. are particularly susceptible to contamination and. as they require more handling.94 (7).0% salt in the water phase (30). The FDA established that hot. 5.5. but. they can survive lightly cured processes when the sodium chloride content is less than 5% and water activities are higher than 0. Sliced smoked fish products. aureus growth. throat and skin. coli all occur on fish products as a result of contamination from animal/human reservoirs. 5. Infection by Shigella sonnei has increased in the UK.86 and 10% sodium chloride. it should contain not less than 3. the human nose. Usually.30% of the population positive for enterotoxin-producing strains (41). gills and guts as well as in several lightly preserved fish 114 . related to the consumption of a raw smoked fish meal.4. the risk of infection can be eliminated by proper cooking. Once the product is contaminated.
6 and water activities below 0. a short period of freezing either of the raw material or of 115 . which include intermediate hosts. and in CT95-1207FAIR). and other local traditional products (7). and the tape worm Diphyllobothrium spp. favours growth of psychrotrophic LAB.2. Until 1997. salt levels of 8 . which considered L.514.95 (45).5 Parasites Despite the presence of parasites in fish being very common.92 . in the EU Fisheries R&D-FLAIR program.8 . Marine LAB is part of the natural flora of chill-stored vacuum packed salmon and cod. pH 4. when an outbreak of nine cases of listeriosis was suspected to have been caused by gravad or cold-smoked trout (47).9. 5. monocytogenes’ in 25 g of the finished product.45 °C. Thus.12%. The round worms. in combination with curing agents and refrigerated storage. SMOKED AND DRIED FISH products such as cold-smoked fish. the only concern to public health is when eating raw or uncooked fish products. researchers are looking for other ways of preventing the pathogen’s occurrence or growth. the USA established ‘zero tolerance’ for the organism. most of them are of little concern to public health. monocytogenes in levels at or above 102 per gram (46).the round worm Anisakis spp. As freezing to -20 °C and maintaining this temperature for at least 24 h will kill all the nematodes. both being detected in cold-smoked salmon. This includes matjes herring. The use of LAB in fish preservation is a topic that has been studied in projects (UP. a number of fish products.5. widely used as starter cultures in food processes. such as ‘lightly preserved’ fish products.CURED. a round worm. They have complicated life cycles. Two outbreaks in the United States have been reported to be linked to salmon (10). are considered unsafe. Pseudoterranova dicipiens. Vacuum packaging (VP) of sliced smoked fish creates a microaerophilic atmosphere. do not survive 1 min at 60 °C. gravad fish. which means that the process must conform to a level of ‘no detectable L. which. marinades (ceviche) (43) and gravad fish (44). from freshwater fish. As they are heat-sensitive. Its limiting conditions for growth are 0 . the CT 3162-FAIR program.0. from seawater fish. lightly salted caviar. cold-smoked fish. there were no reported outbreaks related to cured fish. which means that cooking a fillet 3 cm thick for 10 min at 60 °C will kill any worms present (48). As its presence has been reported in lightly preserved fish products whose processing chain cannot fully assure the absence of the organism. Of microbiological concern in the northern hemisphere are two parasites . monocytogenes pathogenic for only specific segments at risk. or nematodes. Some LAB. have been shown to have an anti-listerial activity in many food products. The Health Protection Agency (HPA) in the UK considered that it was unacceptable for readyto-eat foods (including all the products that are consumed without prior cooking) to contain any serogroup of L. Unlike some European countries. has also been detected in cod (7).
‡ On occasions some strains may be pathogenic.<100 >104 >100 N/A N/A <20 20 .<103 N/A 100 .<100 20 . and unpasteurised yoghurt. monocytogenes) (cfu/g unless stated) <106 106 .100 >100 N/A Pathogens Salmonella spp. the organism should be identified # N/A denotes not applicable 116 . perfringens B. coli (total) Listeria spp. salami. Acceptability is based on appearance. texture. for example.<104 104 .VI Guidelines for the microbiological quality of ready-to-eat smoked fish.<100 200 . monocytogenes Staph. not detected in 25 g Campylobacter spp. E. aureus C. coli O157 & other VTEC Vibrio. † Guidelines for aerobic colony counts may not apply to certain fermented foods.<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. (not L.<107 ≥107 N/A <100 <20 100 . These foods fall into category 5. cereus and Bacillus subtilis# <20 <20** <20 <10 20 . smell.<104 100 .<100 20 .6 Published Microbiological Criteria TABLE 5.<104 present in 25 g ≥103 >100 ≥104 ≥104 <103 103 .FISH AND SEAFOOD the final product should therefore be included in the processing as a means of controlling parasites (7). If the Bacillus counts exceed 104/g. and the levels or absence of indicator organisms or pathogens. 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. 48h ± 2h) Indicator organisms‡ Enterobacteriaceae E. soft cheese. 5.<100 10 .<104 20 .
1997. 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. Van Nostrand Reinhold. Torry Advisory Note No. Urner Barry Publications Inc. 9. in The Seafood Industry. 8. 117 . USA. 334..CURED. Regenstein C. 120-38. Birnie K. Food spoilage fungi. 3273. Introduction to Fish Technology. Van Nostrand Reinhold.F. Hall G. Wareing P. cured and dried fish. monocytogenes Salmonella spp.A. References Regenstein J. Doré I. 7. Eds. 2.. botulinum * Limit of tolerance 1x 103 ETEC/g. New York. 56. coli (ETEC) L. Hot Smoking of Fish. Horne J. Catching.8 ppm (saxitoxin equivalent) 20 ppm domoic acid.W. 254-82 5. Assurance of Seafood Quality. 2007. 37(revised). Chapman & Hall. 1979. parahaemolyticus Vibrio vulnificus C.M. Torry Advisory Note No. Martin R. Wareing P.H.F. Huss H. Leatherhead. aureus * Vibrio cholerae V. Horner W. 6. 1990. * Staph. Marinades. Fernandes R. 1994. Leatherhead Publishing. salting and smoking). 381-406. Bannerman A. Preservation of fish by curing (drying.M. New York. where 30 ppm is permitted Paralytic shellfish poisoning* Amnesic shellfish poisoning * *These criteria are applied to all fish 5.7 1. or presence of toxin 0.E. 82 (revised). Ed. 1993. Smoked. Moody M. 1982. Flick G. 3. FAO Fisheries Technical Paper. Eds..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. SMOKED AND DRIED FISH TABLE 5. in Micro-Facts: The Working Companion for Food Microbiologists. in Fish Processing Technology. London.. Handling and Processing Eels.. Fernandes R. 4. Flick G. 1984. 1991. McLay R. The Smoked and Cured Seafood Guide. Torry Advisory Note No.. except in the viscera of dungeness crab.
20. Embarek P. Dillon R.. Yamauchi H... London. 23.. 51-81. 27 (9). 53 (4). Torry Advisory Note No. Horne J. Doi M... Ninomiya M.C. Technomic Publishers. Shuto Y. 22. 1997. 56 (6). 19. The discovery of umami. Asymmetric reduction of acetophenone by Aspergillus species and their possible contribution to Katsuobushi flavor.. 1989.. 61 (8). Botulism and Fishery Products. Druschitz A. Cann D.. Lindemann B.B.M. Food Reviews International. Food Chemistry. Halpern B. Doe P. 958-60. Eklund M.M. 6 (4).E.. Perusing Asian soup options. Pelroy G.M. 12. Yokayama S. Huss H. 11. Shuto Y.C. Microbiological control for fish smoking operations. 2002. Bioscience.. Bioscience. 108 (3). What’s in a name? Are MSG and umami the same? Chemical Senses. Journal of Food Protection. 1998. 6-dimethoxy-4-methylphenol by Aspergillus glaucus and their possible contribution to Katsuobushi flavour. 1995. Doi M. Kinoshita Y. Doi M. 508.. in Fisheries Processing. 1995. Martin A. Ogiwara Y. 1386-7. 24. 59 (12). Food Control. 14. Oxidation of acetophenone by Aspergillus species and their possible contribution to Katsuobushi flavour.H. 1984.... 843. Shuto Y... 1992. Yajima I.. O-methylation of 2. 1990.W. Bannerman A. Doi M. 21. 18. 18. Kanayama T.. 935-41.. Kinoshita Y. Inhibition of Clostridium botulinum types A and E toxin production by liquid smoke and NaCl in hot-process smoke-flavoured fish. Teeny F. 2324-5. 335-40. 22 (revised). 1980. Peterson M. Food Product Design. 15. Packing Fish in a Modified Atmosphere. Aoshima H. 25. Sakakibara H. Cann D. Hosokawa M. Biotechnology and Biochemistry. 1981... Biotechnology and Biochemistry.. 2008. Biotechnology. Ed. 17.. Fish Drying and Smoking: Production and Quality. 1977. Ninomiya Y. Agricultural and Biological Chemistry. 2007. 13. 840-6. Control of biological hazards in cold smoked salmon production.E. Lancaster. Shiemori Y. Paranjpye R. Jeppesen V. Torry Advisory Note No.A. 6 (6). 88. Mitou M... Bioscience. 1994. Flavour constituents of dried bonito (Katsuobushi). Degradation and omethylation of phenols among volatile flavour components of dried bonito (Katsuobushi) by Aspergillus species. Matsui M. Hayashi K. Bannerman A. Processing Cod Roes. and Biochemistry. Patel T. 1051-5. 553-72.. Torry Advisory Note No. Recommendations for the Preparation of Smoked Salmon. 1982. Matsui M. Martin A.R. 5 (revised). 17 (9). Torry Advisory Note No. 118 .F. Effect of dried bonito (Katauobushi) and some of its components on GABA-A receptors.K. 45 (10)..P.FISH AND SEAFOOD 10. Chapman & Hall. Biotechnological Applications. 16.
Luxembourg. 22. Hocking A.K. in Fish and Fishery Products Hazards and Controls Guide. 1995. The microbiology of sea-water fish.A.T. 351-7. Lancet.. Smoke flavourings. CEC. Cox J. The control of botulism in hot-smoked trout and mackerel.. 1991. 1986.J. Institute of Medicine. 1994.F.. 42. 348. in Smoke in Food Processing. Washington DC. 43.. 30. Springer Science. Commission of the European Communities.. Golden D. Food Technology. Hocking A. London.. 27. Maga J.. 1961. 1996. Borgstrøm G. Press Inc. in Reports of the Scientific Committee for Food (34th Series). 34.H. US Food and Drug Administration. Journal of Tropical Medicine and Hygiene.CURED. 31. Shewan J. Pitt J.I. 1971. 29. 28. Studies on Sporendonema epizoum from ‘dun’ salt fish. Incidence of Listeria monocytogenes in an acidified product..A. Tour highlights production and uses of smoke-based flavours. Blackie. 74 (2).I.M. 70-2. Food Research International. Fungi and Food Spoilage. Townsend J. Food and Drug Administration. 311-7. Ed.M. An outbreak of Shigella. Oxford.S.A. Food and Drug Administration. Pitt J. Food Microbiology. 1993. Ed. 37. Adebajo L. SMOKED AND DRIED FISH 26. Fuchs R.C.I. 1941. Wheeler K. Rockville. Hansen L. 1979.1988. 5. 119 .D. Taylor L. 74. Ed. National Academic Press. 1996.V. 3 (4). Huss H. 181-8. Lucas F. Journal of Food Technology. Antimicrobial properties of smoke. Journal of the Fisheries Research Board of Canada. 2005. 36. Smoked and smoke-flavoured fishery products. 218-25.F. Jay J. Pszczola D. Letters in Applied Microbiology.D. 1991. V. Committee on Evaluation of the Safety of Fishery Products. 742. Academic Press.Y. 40. 41. CRC.E. Seafood Safety. Anggawati A. 14. 27686. Sprouse R. Mycotaxon.. 1995. Gill T.D. 197-208. Ahmed F..M. ceviche. Pitt J.. 123-9. Scientific Committee for Food. 1997... 88-90. Frank M. Hess E. 1985.R.O. 33. 12. in Fish as Food.E. Effects of sodium chloride and relative humidity on growth and sporulation of moulds isolated from cured fish. 1994.A. 32. Journal of Applied Bacteriology. Studies on salt fish. Wheeler K.. Cann D. New species of fungi from Indonesian dried fish. 38 (3).D.B. 39. Nahrung. 29 (2). Fungi associated with Indonesian dried fish. Loessner M. Importance of autolysis and microbiological activity on quality of cold-smoked salmon. Diyaolu S. 38. New York. Boca Raton.. Simini B. Ed. Sirvas S. Røntved S. Interactions among xerophilic fungi associated with dried salted fish.. 98100. Fungal flora of South Vietnamese fish and rice.. 49 (1). Hocking A. 164-9. 35. Modern Food Microbiology. 74. 107-11... Hocking A.
3.. Acta Veterinaria Scandinavia. 507-10. 5 (1). Garren D.. Loncarevic S. Amsterdam. Journal of Food Protection. Unnerstad H.. Cann D. Hughes A. Richards J. Cured.. 1996. Moral A. 80. 2000. Bolton F.A. Paranjpye R. 1980. Loncarevic S. 47. Microbiological guidelines for some ready-to-eat foods sampled at the point of sale. 5-7. 1991. US FDA Centre for Food Safety & Applied Nutrition. 26 (4). Liston J. http://www. Growth and production of toxin of Clostridium botulinum type E in rainbow trout under various storage conditions. in Quality Assurance in the Fish Industry. Persson I. 48. Nye K.. Jacobsen M. 45.. 49-59. Inhibition of Clostridium botulinum types A and E toxin formation by sodium nitrite and sodium chloride in hot-process (smoked) salmon. Ed. Zeitschrift für Lebensmittel-Untersuchung und –Forschung. 45 (9). 5.. 232-6. Journal of Clinical Microbiology. US FDA Centre for Food Safety & Applied Nutrition. Harrison M. 1997. Eklund M. 46. de Louvois J. Ericsson H. 1982. Food Control. 863-6.. salted...S.R. 197-209. Clostridium botulinum and its importance in fishery products. 37 (1).html Gilbert R.. Journal of the Association of Food and Drug Officials.. Mentzing L-O. Tham W.. 1995.M. Journal of Food Protection. 1992. 13-8. Keeping quality of vacuum packed smoked sardine fillets: microbiological aspects.D. Roberts D. 120 . in Fish and Fishery Products Hazards and Controls Guide. 135-85.J.W. Ribeiro C..8 Further Reading Gibson D.J. Washington D.N.L.. Hurst A. Round Worms in Fish. The effect of NaCl on the upper temperature limit for growth and enterotoxin synthesis by Staphylococcus aureus. Beltran A. Fletcher R.. Wooten R.gov/~comm/haccp4x5. Prevalence of Listeria monocytogenes and other Listeria spp.C. Association of Food and Drug Officials. Tham W..fda. Huss H. Danielsson-Tham M.FISH AND SEAFOOD 44.. in smoked and "gravad" fish. Eklow A. Huang Y.. and smoked fish establishments good manufacturing practices. 833-41. Food and Drug Administration.. 35 (11) 2904-7. 22.cfsan. Canadian Journal of Microbiology. Pathogenic microorganisms of importance in seafood.M.V..W. Advances in Food Research. Little C. Pelaez C.A. Danielsson-Tham M-L.. 1989.. 1976. Hobbs G.C.. Pelroy G. Donovan T. 55 (4). Incidence and growth of psychrotrophic Clostridium botulinum in food. Davies A. Gibbs P... Torry Advisory Note No. Peterson M.. 188 (3).A. Eds.E. Appendix 5: FDA and EPA Guidance Levels. 1980.. 1994.. 163-7. 58 (8). Elsevier Science Publishers B. Susuki E..H. 2001. An outbreak of listeriosis suspected to have been caused by rainbow trout. Communicable Disease and Public Health.
27 (6). 217-36. 14. 1734. pH and sodium chloride. Ben Embarek P. in Seafood Quality Determination. Occurrence of Listeria monocytogenes in freshwater farms and fishsmoking plants. Paranjpye R. Food Technology.A.. Eds.T. Incidence of Listeria in hot and cold-smoked fish. 19. 25 (5).A. Presence. 1974. 1994. 11. Lashbrook L... 23 (1). Efficacy of high sodium chloride concentrations for the destruction of Listeria monocytogenes. 1973. 1992.. 844-6. Keusch A. 11.S. van Spreckens K.. Rørvik L... Prevalence of Listeria in smoked fish. solute. Bayliss A. Yndestad M. in a salmon slaughterhouse and smoked salmon processing plant. Davies A. Interacting effects of pH. Roberts T. 309-18. Elsevier. Tran T. 25. Hitchins A. 1995. Histamine production by psychrophilic flora. Journal of Food Safety.. 94-8.K.. Dallas H.J. New York. Caugant D.178-80. 1994. Ratnam S. Contamination pattern of Listeria monocytogenes and other Listeria spp.. Competition of food bacteria with Listeria monocytogenes during enrichment culture.W. 61 (12). SMOKED AND DRIED FISH Sutherland J. Letters in Applied Microbiology. temperature. 293301. Fuchs R. and salt concentration on growth and survival of Vibrio parahaemolyticus. Hudson J. Poysky F. Taylor S.R.E. 121 . Combined effects of water activity. Letters in Applied Microbiology..CURED. Eklund M. Patel T. 1994.N. International Journal of Food Microbiology. Beuchat L. Slade A.R. Journal of Food Protection. Peterson M. 19-27. Letters in Applied Microbiology. Predictive modelling of growth of Staphylococcus aureus: the effects of temperature. 1075-80. 4389-95. Poindexter C. 502-8. Beuchat L. detection and growth of Listeria monocytogenes in seafoods: a review. Pelroy G. 42... Applied Microbiology. Food Microbiology.R. Jemmi T. 58 (5). 1995. 1995. 1988. Applied Microbiology. and temperature on the growth of Vibrio parahaemolyticus. 309-16. Fate of Aeromonas and Yersinia on modified-atmosphere packaged (MAP) cod and trout. 55 (11). Marine toxins of microbial origin. 1994.. Nicolaides L... 1991. Dillon R. International Journal of Food Microbiology.. Journal of Food Protection. Incidence and sources of Listeria monocytogenes in cold-smoked fishery products and processing plants. 394-6. 1992. International Journal of Food Microbiology.. Kramer D.A.C. 1987. Liston J.P. 866-70. 21 (3).
As with most traditional products that are still produced principally on a cottage industry or domestic scale. there are numerous variants of some common themes and a host of local names used to describe them. they are exported to European.1.1 Definitions As applied to fish products. Martin Adams Faculty of Health and Medical Sciences University of Surrey Guildford Surrey GU2 7XH United Kingdom 6. the most important of all fermented fish products. It is produced from the sandfish Arctoscopus japonicus and has the rice-based enzyme preparation. North American and other markets and are. For example. typically in the range 15 . The relative importance of these two activities depends on the formulation of the product. 123 . they can be divided into two categories: fish/salt products and fish/salt/carbohydrate products. the best-known of the fish sauces (gyoshoyu) of Japan is shottsuru. Sugar can also be added to the Indonesian fish sauce bakasang and the Thai product nam-budu. producing the fish sauces and pastes. Fermented fish products are largely confined to east and south-east Asia. to those processes in which the activity of lactic acid bacteria (LAB) plays an important part. lactic fermentation can play a significant role. in economic terms.6. whereas in those where a carbohydrate source such as rice or sugar is also added. Essentially.25% and are used mainly as a condiment. the term ‘fermented’ describes a spectrum of processes that ranges from the largely autolytic degradation of fish protein. though some are produced elsewhere. added during preparation. 6. The fish sauces are produced on the largest scale. Those to which only salt is added tend to be mainly autolytic. FERMENTED FISH Prof.1 Fish/salt products such as the fish pastes and sauces tend to contain relatively high levels of salt. The principal fish sauces and pastes of south-east Asia and their local names are listed in Table 6. Carbohydrates are added to some fish sauces during their preparation. koji.I.
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. who had adopted them from the Greeks. a variety of products can be produced within a single country. salty taste.2 Fish/salt/carbohydrate products range from those that resemble the fish sauces and pastes in the sense that extensive autolysis has occurred. cheese-like aroma Burma Indonesia Ngapi ketjap-ikan Paste Red/brown salty paste nga-ngapi trassi-ikan trassi-udang (shrimps) prahoc mam-ruoc (shrimps) Kampuchea nuoc-mam nuoc-mam-gau-ca (livers only) Korea myulchljeot jeots. and the more widely known Worcestershire sauce a very distant relative. may be direct descendants.1. A selection of Asian products is presented in Table 6. jeotkals (shrimp) nam-pla (pa) Budu Patis nam-pla nuoc-mam padec Belachan (shrimps) bagoong kapi mam-ca man-tom (shrimps) Laos Malaysia Philippines Thailand Vietnam Even within the limitations of fish and salt as the sole ingredients. and pissala. to products analogous to other lactic fermented foods such as salami.I Fish sauces and pastes of south-east Asia Country Sauce Amber/brown liquid. In Europe.II. where the bacterial production of lactic acid is a major feature. although garos.III. These have long since fallen from use.FISH AND SEAFOOD TABLE 6. made from fish livers in Greece. Fermented fish products are of very minor importance outside Asia. 124 . 6. made from whole fish in France.
pla-chao. mam-chao. som-pa.12 months 10% raw sugar added. crustaceans Mackerel eviscerated fish Variety of fish Whole 1:4 Condiment Pla-thu-khem Whole 1:3 2 . som-pa-keng pekasam. shrimps.FERMENTED FISH TABLE 6. e. pa-khem.III Fish/salt/carbohydrate products of south-east and east Asia Country Japan Kampuchea Korea Laos Products I-sushi. burong-bangus pla-ra. e. burong-ayungi. crustaceans Variety of fish Variety of brackish or marine fish 1:4 3 .II Fermented fish/salt products common in Thailand Name Type of fish Form Approx. funa-suchi. ayu-sushi.4 months Condiment and main dish Whole 1:3 18 months Condiment Whole 1:3 After 3 . cencalok burong-isda.g. pa-chao. tai-suchi phaak. som-fak/ som-fug 125 Malaysia Philippines Thailand . burong-dalag.g. pla-som. mam-pa-kor.3 months Main dish Tai-pla Bowels 1:3 6 . salt/fish ratio Production period Type of use Hoi-dong Molluscs Whole or without shells Whole - Main dish Kapi (fish or shrimp paste) Nam-pla (fish sauce) Nam-budu Small fish. the mixture is then boiled and bottled - Condiment Nam-khoei Shrimps.8 months Main dish TABLE 6. mam-seeing sikhae som-kay-pa-eun.
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. Here. Table 6.FISH AND SEAFOOD In Northern Europe. This could lead to quite marked variations since the same fish can be obtained from a variety of sources. which employ lower levels of salt than Asian products. the most popular varieties of which are rakørret. for example. which has been increasing in popularity over the last 50 years. relatively mild-tasting product.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. 6. fermented trout. TABLE 6. and their microflora tends to reflect their local environment more than that of marine species. in Norway and surstrømming. malt or flour (1). is a traditional. A modern account of their production. 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. and old descriptions of their production often refer to the addition of whey. paddy ponds beside rice fields used for collecting fish when the field has dried up. however. especially in fish/salt products.IV) and in the way in which they are handled prior to processing means that the initial microbial levels are far from uniform. freshwater fish are the usual raw material. Penaeus indicua Many of the fish used in fermentation are uneviscerated. it was found that several different sources were used for the fish . rakefisk or surfisk. in a survey of fermented fish products in north-east Thailand. some fermented fish products remain popular. and counts ranging from below 104 /g to in excess of 107 have been reported. in Sweden. states that sugar is often included with the salt. made from herring. For example. The diversity of fish used in fermented products (see. There are also more heavily fermented products. or buried salmon. and a large local freshwater reservoir (2). Norwegian gravlaks. 126 .the flooded rice fields. These are often described as fish/salt products. and the initial microflora is likely to be considerably larger in these cases.
Generally. 6. the product contained about 500 cfu/g comprising mainly Micrococcus and Bacillus spp. principally species of the genera Rhizopus.1 – 6. work with nam-pla in Thailand has isolated Halobacterium and Halococcus spp. Other halophilic species have been identified from fish/salt products. Mucor. In some cases. in many cases. It also demonstrated their significant proteolytic activity during the first month of fermentation. It has also been shown that a perfectly acceptable patis could be produced from fish that had been sterilised with ionising radiation (6). suggesting that halophilic bacteria do play an important role in the production of fish sauce (8). were found to be the predominant isolates.organisms that play an important part in the production process but can also contribute to the ultimate spoilage of the product. in numbers reaching 108 /ml after 3 weeks of fermentation but declining thereafter. mean that microorganisms have been generally held to play little or no role in the process that follows. the Philippine fish sauce. however. Bacillus spp. where the total counts dropped from above 107 cfu/g to below 103 cfu/g after 14 days and below 102 after 40 days. 4). Similar results were seen with patis. More recently. its contribution is likely to be insignificant compared with that of the fish microflora in terms of both spoilage potential and public health concerns. Similarly microbial inactivation by irradiation has not been found to adversely affect the properties of Korean fermented squid (7). however.74 or below. probably reflecting their ability as spore-formers to survive rather than any capacity to multiply under the prevailing conditions (5).3 6.4.1 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. Studies have shown that the number of bacteria declines rapidly during the production of the Thai fish sauce nam pla and that. after one month. traditional starter culture preparations such as koji (Japan). 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. In another microbiological study of four fish sauces. Hansenula.3. 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. These preparations contain a mixed microflora of moulds and yeasts. Endomycopsis and Saccharomyces . This.FERMENTED FISH Where other ingredients are used. and studies 127 . which included nam-pla and patis. and the anaerobic conditions that prevail in the bulk of the material. When this is a simple carbohydrate source such as sugar. (3. sufficient to saturate the water phase present and produce an aw of 0. angkak (Philippines and Thailand) and look-pang (Thailand) are used. either directly or as the partially saccharified rice they produce. or boiled or roasted rice. they can contribute to the initial microflora.
Fig.FISH AND SEAFOOD have demonstrated the production of halophilic proteinases from fish sauce isolates (9 . 6.1. Belacan (Condiment) Fig. 6.2.12). Patis (Condiment) 128 .
thought to be largely the result of autolytic breakdown of the fish tissues. To produce semi-solid fish pastes as opposed to fish sauces. In fish sauce production. the 129 . 14). Myulchijeot (Staple food) The extensive proteolysis and liquefaction that occur during production are.4.3. and muscle cathepsins have been shown to be inhibited at relatively low salt concentrations (13.6. however. 6.FERMENTED FISH Fig. eviscerated fish are sometimes used to slow the autolytic process. 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.
Some examples of products and their formulation are presented in Table 6. In contrast to free-swimming fish. and the low pH produced contributes a second protective barrier or hurdle against the growth/survival of the normal fish-spoilage organisms and pathogens.5). filtered and bottled.5.2 Fish/salt/carbohydrate products of Asia This category encompasses an even greater diversity of products than the fish sauces and pastes. Their main feature is that the provision of a source of carbohydrate allows a lactic acid bacterial fermentation to occur.FISH AND SEAFOOD high salt level used also assists in the osmotic extraction of a supernatant rich in amino acids. 6.V and Figures 6. 6. Hoi-Malangpu-Dong (side dish. Fig. Figure 6.3.6 – 6. particularly glutamate. which is eventually separated from the residual fish tissues.8. snack) 130 . volatile fatty acids and nucleotides. shellfish such as mussels often have appreciable levels of carbohydrate in their tissues and in these cases addition of carbohydrate is not necessary for a lactic fermentation to occur (see. for example.
C NE.12 months Shelf life Use 1 .7 days 5 . Ang-kak: red mould rice obtained by fermenting rice with Monascus purpureus. north. north-east. N S 5 .V Fish/salt carbohydrate products of Thailand Name 1 Pla-ra Region C. Fig.5:2-3:1 Fish:salt:boiled rice: ang-kak 3:1:3:0. C NE. In these cases. the prefix 'pla' is replaced by 'kung'. 3:1:0. S. 3:1:1-3 Fish:salt:boiled rice: garlic 10:2:1:0. N.6. NE Formulation Fish (whole or pieces): salt: roasted rice.6.4 are also produced using shrimps. south of Thailand. NE.20 days 3 Pla-som 4 Pla-jom 5 Som-fak 6 Pla-paengdaeng NE. NE 10 .03 Preparation time 6 . C.FERMENTED FISH TABLE 6.12 days 3 . side dish) 131 .12 months Main dish 2 Pla-jao C. Kung-Chom (staple food. N.5-1.10 days 5 days Main dish Main dish Main dish Condiment and main dish Note: Products 1 .25-1 Fish:salt:roasted rice: garlic 10:1:3:1 Fish (minced):salt:rice: garlic 10:0.2-4 Fish (pieces): salt:khaomark (fermented rice). N. central.3 years Condiment and main dish 2-3 months 3 weeks 2 weeks 2 weeks 6 .
6. their growth and metabolism are nonetheless slowed. 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 6.1 The use of higher salt levels results in a longer production phase but a better keeping quality product.VI). It also indicates that. 2. then this will take longer to develop than if lower salt levels are used.7. The LAB isolated from these products are those with more marked salt tolerance but.salt and lactic acid. but there is considerable variation in this.10%. while they are able to grow at high salt levels.. Phillipines Journal of Food Science and Technology. Red burong isda 132 . giving a reduced rate of acid generation. Generally. If acidity is a desired sensory property of the product.FISH AND SEAFOOD The extent to which lactic fermentation occurs depends largely on the salt content. (from Arroyo et al. A survey of fish/salt/carbohydrate products in Thailand (15) noted that there were two empirical rules governing formulation:6. typically in the range 2 . the salt is of far greater importance in determining the product’s long-term stability. of the two factors contributing to shelf life . This is readily explained by the susceptibility of LAB to elevated salt concentrations.2.3. these products are salted at lower levels than the fish sauces and pastes. 106) Fig.
0 .1 .17.9 4.5 3.9.3 .5.6.0 Shelf life 3 weeks 6 .5.2 7.6 3.FERMENTED FISH (from Arroyo et al.6.4 .5. 133 .5 .5.8.9 4.2 4. White balao-balao TABLE 6.0 . Philippines Journal of Food Science and Technology.8 pH 188.8.131.52 4..6.2 4.0 . 2.8 2.3 years 1 .5 .4.VI Shelf lives reported for fish salt carbohydrate products Product Pla-som Pla-paeng-daeng Pla-ra Pla-chao Pla-chom Som-fak Salt 2.7 .3 5.12 months 1 .9.2 years 2 weeks 2 weeks Source: A Concise Handbook of Indigenous Fermented Foods in the ASCA Countries (16).8 . 106) Fig.8 .9 .
VII Fish/salt/carbohydate products in which saccharified rice or a saccharifying starter culture is used Product red burong-isda i-sushi.VII). For example.3. Although the ability to ferment starch is not widespread in the LAB.4.1 . Rice is the most common carbohydrate source used. TABLE 6. The effect of this will be to decrease the product’s final pH and increase the amount of total acidity. 21).0 . they increase the amount of soluble sugars produced and thus the range of LAB that can grow. Addition of carbohydrate will. a survey of burong-isda in the Philippines reported consistently lower pH values for red burong-isda where ang-kak was used (pH 3. efficient lactic acid production will depend upon the presence of amylolytic LAB or saccharification by other microorganisms or enzymes that may be present. In several products. When added as boiled or roasted rice alone. 19). alternatively. 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 (20.2. the rice may be added in a partially saccharified form such as kao-mark (Thailand).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.FISH AND SEAFOOD 6. inulin. being similar (18.9) compared with the white product (pH 4. of course. also serves as a valuable source of the more readily fermented carbohydrate. 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. also lower the unit cost of the product and this is probably a significant factor in many cases.5) despite other factors. Where they are used. a common ingredient in these products. This results in a more active lactic fermentation and a lower pH. 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). although the final population of LAB may be similar. Garlic. a saccharifying agent such as koji (Japan) or ang-kak (the Philippines) may be added separately (Table 6.3.
around 12%. red pepper powder and garlic in the ratio fish:millet:pepper:garlic of 80:7. 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. 135 . which ranged from 7. reporting the product’s pH to range from 4. but they are likely to behave similarly to lactic fermented products produced from other commodities. A combination of irradiation and chill storage has been shown to prevent both over-acidification and fungal growth in som-fak. but in most cases it is probable that yeast and mould growth would be an important cause of spoilage. will not reliably exclude air. thereafter declining to about 4. such as wrapping in banana leaves. Traditional packing techniques used with lactic fermented fish products. the level of salt employed is on the high side for fish/salt/carbohydrate products. Clearly. In a successful fermentation.8 .3 weeks at around 20 °C.7 up to 6. Generally. and there is a marked regional variation in the preferred type.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). and surface mould growth has been shown to be a cause of rejection in the Thai product som-fak (24). markedly extending the shelf life (25). The growth of yeasts and moulds could. It seems that a lactic fermentation takes place in many types of pla-ra but is not significant in others. in turn. which is sufficient to prevent lactic fermentation by all but the most halotolerant LAB. The Thai product pla-ra exemplifies these basic rules as well as the difficulties associated with attempts to classify such products. lead to an increase in pH. The mixture is allowed to ferment for 2 . the pH drops from 6.7 to below 4. This suggests that use of oxygen-impermeable wrapping films could extend the shelf life of such products.9%.2 at the end of fermentation. Excessive growth and acid production by the LAB could render products too sour for acceptability.3 days.2.17. 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). presumably correlating with the salt content. Any loss of acceptability must arise from chemical or physical changes occurring over prolonged storage. the Korean fermented fish product. the salt levels in the latter are sufficient to rule out microbiological spoilage in all but the most exceptional of cases.5. There is little published information on the factors that limit the shelf life of the low-salt products. Cleaned pieces of eviscerated fish flesh are salted with 6% salt overnight before mixing with cooked millet. One study found that any effective measure to exclude air from a fermenting fish/carbohydrate mix prevented yeast growth (23). Results of a product survey were consistent with this. allowing other spoilage organisms to grow. is something of an exception since millet is used along with garlic as the carbohydrate source.5 in the first 2 . 6.5:9:3.FERMENTED FISH Sikhae.
After capture. parahaemolyticus has been reported growing down to a pH of 4. and consumed without cooking. but we are unaware of any reports of this happening.8 and at salt levels as high 10%. under conditions inimical to growth and high ambient storage temperatures. viable numbers will probably decline quite rapidly. the pH decreases below 5 and safe processing is recognised to depend on proper salt levels and temperature control until the pH drops. fish products are the most common source of botulism.5 Pathogens: Growth and Survival Many fermented fish products are stored at ambient temperatures. V. and the most frequently implicated product is the fish-salt-carbohydrate fermented product Izushi. The epidemiological evidence points to greater awareness of the importance of these factors since outbreaks have declined over recent years (27). In the fish sauces and pastes. fish can potentially be contaminated with almost any foodborne pathogen as a result of improper handling and storage. Their safety therefore is critically dependent on the quality of the raw materials used and the inhibitory effects of the fermentation process. In Japan. 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. although these limits apply when all other factors are optimal (26). Improper storage of fish prior to processing could result in the production of toxins capable of persisting through to the product. There is little in the way of documented information associating fermented fish products with foodborne illness. Because of the severity of the illness it causes. however.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. are Vibrio parahaemolyticus and Clostridium botulinum. During fermentation.87. such as soaking the fish at temperatures below 5 °C before fermentation and the use of starter cultures to ensure a rapid and reliable decrease in pH. and failure to achieve a sufficient combination of inhibitory factors could lead to their growth or survival. As in Norway. The fish/salt/carbohydrate products depend on the combination of salt and pH/lactic acid for their safety. The bacterial pathogens most commonly associated with the living environment of fish. outbreaks of 136 . The Scandinavian fermented fish product rakefisk was responsible for 16 (84%) of the outbreaks of botulism recorded in Norway in the period 1961 90. Several measures have been suggested for reducing the risk of botulism. 6. The combinations of pH and salt in many fermented products are likely to preclude growth of the organism completely and. typically 30 °C. which accounted for 88% of outbreaks in the period 1952 .
though 27% survival was still recorded (33). One study with the trematode Haplorchis taichui. 137 . beaver tail and paw. Novus Press. Many of these products are described as fermented. which is generally consumed raw (29). Saisithi P.. Botulism has also been associated with a number of traditional foods of the peoples of northern Canada. They are packed into containers such as buckets or pits in the ground and left at ambient temperature to putrefy. and salmon heads and eggs. Baseline Study (1987-1988). Journal of Food Science. Infection with the trematode Opisthorchis viverrini is associated with cholangiocarcinoma. botulinum.O.. Thailand. 31. 1966. Where these products are consumed without final cooking. showed that inclusion of fermented fish in the Thai dish lab-pla improved inactivation of the metacercariae.. Greenland and Alaska. although the absence of any source of fermentable carbohydrate means that the process is putrefactive.. Salting levels are often insufficiently low to prevent growth of C. viverrini were a strong risk indicator and that consumption of fermented fish products was an independent risk factor in the contraction of cholangiocarcinoma (31).M. while increasing the acetic acid content of the formulation had little or no effect (32). 2. 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). Liston J. Foodborne trematode infections are emerging as a major public health problem. Kasemsarn B. and these are often used in the production of fermented fish products.. References Riddervold A.FERMENTED FISH botulism have become less frequent in recent years. botulinum and the production of toxin. which will persist into the product. Mahidol University. they could act as an important vehicle of transmission. Dollar A. giving an alkaline fermentation. Fermented Fish Products Study (Thailand) Phase 1. The more widespread current use of non-traditional containers such as glass or plastic jars is thought to be one contributing factor (28). probably reflecting greater awareness of the critical factors that determine the product’s safety (27). Puwastien P. 1989. affecting an estimated 40 million people. particularly in south-east Asia and the western Pacific regions (30). Yhoung-Aree J. Oslo. Rakefisk and Herring in the Norwegian Tradition. Microbiology and chemistry of fermented fish. With this particular organism. Institute of Nutrition. 6. salt content was found to have the greatest impact on the viability of larvae. Dhanamitta S. 105-10. Raw materials subject to this process include seal flipper. 3. whale meat. walrus nose. and warm weather can result in rapid growth of C. Freshwater fish are the most important intermediate reservoir hosts of foodborne trematodes.6 1. An investigation of survival of the fish parasitic nematode Anisakis in marinated herring has given some indication of what these studies might find. A case-control study conducted in northeast Thailand showed that elevated levels of antibodies against O. 1990. Lutefisk.
Thawai C. 3649. 8..T. Tsunemi M.. Kyong-Haeng L. Ono H. in General and Applied Aspects of Halophilic Microorganisms.FISH AND SEAFOOD 4.A.. Lentibacillus kapialis spp. 6.. Government of Australia. 7.. 124-30. Nov. Moonmangmee S. 63. Sinsuwan S..M. Microflora of four fermented fish sauces. 643-7. Journal of Fermentation and Bioengineering. 61-73. Journal of Food Science and Technology. 1995. Plenum Press. 106-8. 13. Journal of Food Protection.. 15. 29. Hiraga K. 5..K. Crisan E. Takada K. 353-63. 17-30. Rattagool P. Chung Y.W. 2006. 43. Backhoff P. Bioscience. Saono S.H. A halophilic serine proteinase from Halobacillus spp. Halophiles in Thai fish sauce (Nam pla). Kaneko Y. 1957. SK33 isolated from fish sauce fermentation.. Journal of Scientific and Industrial Research (India). Yook H. 141-2. 2007. Lee S.. Namwong S. Constantinides S. 1395-401. isolated from jeotgal. 57. Journal of Food Science. Fukuda H. Characterisation of starch-hydrolysing bacteria isolated from a fermented fish and rice food “burong isda” and its amylolytic enzyme. New York.. Catheptic activity of fish muscle. Agents of proteolysis and its inhibition in patis (fish sauce).H. Lim J. 9. SR5-3 isolated from fish sauce: purification and characterisation. Canberra. Production and characterisation of NaCl-activated proteinases from Virgibacillus spp.K. International Journal of Systematic and Evolutionary Microbiology. 16. Tropical Science. 57. Journal of Food Technology. Applied Microbiology.R. Suntinanalert P. 1985. Reddi P.R.. Orejana F. 11. Hull R..J. Some chemical changes in fish silage. 934-9.M.... Velankar N. Nov. Kudo T... 1991. 17. Aslam Z. a traditional Korean fermented seafood.. 2008. 2007. 6338. Dymsza H.S. Salinococcus jeotgali spp. Oda K. Tanasupawat S. Rodtong S.D.T. 138 . Cooke R.R. Ed. Jae-Hun K.. 16A.. 12.. 1979.. Biotechnology and Biochemistry. Phil. Im W. 2000. Adams M. from fermented shrimp paste in Thailand... 1975.. Sands A. 1972. 185-92. Olympia M.. Protein hydrolysate from fish. Process Biochemistry. microbiological and chemical properties of salted and fermented squid.. 1986.. 80.. Dong-Ho K. Ahn H. 37. Fermented fish products of south east Asia. Liston J.. 14. 25. Byun M. Itoh T.V. 381-8. Rodriguez-Valera F. International Journal of Systematic and Evolutionary Microbiology. 3. Tanasupawat S. Yongsawatdigul J.. Thongthai C. Pakdeeto A. Takano M. Yasir M. 70. Dhamcharee B. Effects of gamma radiation on sensory qualities... 11.. 10. A Concise Handbook of Indigenous Fermented Foods in the ASCA Countries.. 1976..
139 . 24. Riebroy S.H. Blackie. Wedell-Neergaard C. 27.B. 19. in Microorganisms in Foods. Rustad T. 2002. Sakai H.L. Hazards from northern native foods.. Volume 5: Microbiological Specifications of Food Pathogens... Eds. Benjakul S.. 21... Journal of Applied Microbiology.. Huss H.. 104-6. Owens J. 1815. a Thai low-salt fish product. ASEAN Food Journal. Eds. 69-104. New York. 28 (6). Tanaka M.H. Gram L. Caldo G. Fermentation and spoilage of som fak. Caldo G. 27. New York. a Thai low-salt fermented fish product. Canada Communicable Disease Report. 22... 1983.. Wainwright R. Ed. 20. Epidemiology of human foodborne botulism.A. Journal of Agricultural Science (Tokyo). Hauschild A. 2007.W. Hauschild A.. London..FERMENTED FISH 18. Vibrio parahaemolyticus. Saisithi P. Two outbreaks of botulism associated with fermented salmon roe – British Columbia. The influence of oxygen accessibility on the growth of yeast in fish/rice fermentation. Paludan-Müller C.. Patrick D. Huss H. 3-10. 1995. 92.. Valyasevi R.H. Tropical Science.. Traditional fermented fish products with special reference to Thai products. Control of foodborne trematode infections.. Dodds K.M.. 28. The fermented fish food. Effect of irradiation on properties and storage stability of Som-fug produced from bigeye snapper.W..B. 105-12. Yeast-flora in red burong-isda a fermented fish food from the Philippines. Isaac-Renton J.. 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.H. Gram L. Kozaki M. Genotypic and phenotypic characterisation of garlic-fermenting lactic acid bacteria isolated from som-fak. 1-4. 103. in Clostridium botulinum: Ecology and Control in Foods.. 2002. Sakai H. Avurhi J. burong-isda.L. 1999. Martin J. in the Philippines.H. Hauschild A. 38. 1987. Marcel Dekker. International Commission on Microbiological Specifications for Foods.K. 1992. Moody L. Yamprayoon J. 1990.. 219-29. 1983. Dawar M. 1996. in Clostridium botulinum: Ecology and Control in Foods. 29.A. 305-22. Journal of Food Science and Technology. 426-35. 28.. 25. Food Chemistry. Kozaki M. 1998. Journal of Agricultural Science (Tokyo). 23... 26. Erikson U. 28.W. World Health Organisation Technical Report Series. 30. Marcel Dekker. 3. 1992. Huss H. 138-44. Visessanguan W. Fung C. International Commission on Microbiological Specifications for Foods. International Journal of Food Microbiology. Dodds K.D. WHO Study Group. Ben Embarek P.H. Paludan-Müller C. Gram L. 46. Ostergaard A.D. 307-14.
6. Kikukawa H. Huss H.H. International Journal of Food Science & Technology.FISH AND SEAFOOD 31. Adams M.. Srivatanakul P. Miwa M. 23. Jedpiyawongse A. International Journal of Cancer. 36.. Progress in Industrial Microbiology... 2005. 32.. Fermentation and Food Safety. Wongsawad C.7 Further Reading Adams M.R..R.. Nout M. Fermented flesh foods. Survival of Haplorchis taichui metacercariae in lab-pla. Sriplung H. 854-60. 1986. northeast Thailand.. Horst K. Uchida K. Chuboon S. Ruamsuk A. Roepstorff A.. 29. Southeast Asian Journal of Tropical Medicine and Public Health.J... Genetic and environmental determinants of risk for cholangiosarcoma via Opisthorchis viverrini in a densely infested area in Nakhon Phanom.. Gaithersburg... Thai traditional food preparation.. 110-1. Survival of Anisakis larvae in marinated herring fillets. Honjo S. 661-70. Sripa B. Hanai S. 117. Bloemsma B.R. Deerasamee S. 140 . 33.. 2001. Nithikathkul C. Aspen Publishers Inc.. 15998.. Kittiwatanachot P. Todoroki T. 1995. 2005.
C. Ph. While some of these toxins only occur in seafood from a restricted geographical region. the export of exotic seafood that may contain such toxins can lead to unexpected public health issues. Since the original chapter in this series was published (6). including the well known marine toxins 141 . Liquid Chromatography/ Mass Spectrometry/ Mass Spectrometry -LC/MS/MS) is employed to identify and confirm the presence of known toxins. C. more traditional methods such as the mouse bioassay continue to be used in many countries to warn of the presence of toxins. Wright. Nevertheless. 5).1 Introduction Shellfish and finfish toxins continue to be a threat to public health and local fisheries around the world. several scientific reports have described their efficacy and potential. known and unknown.M. a number of new toxins and toxin families have been identified. An emerging technology is the use of immunosensors (1 .. Increasingly sophisticated instrumentation (e. Another book dealing with the clinical effects of a variety of poisons and toxins.7. It remains to be seen if this approach gains momentum in the future..3) and some practical applications for detecting shellfish toxins have been reported (4. The manual on harmful marine algae and their products. most notably the azaspiracids and the cyclic imines as well as some additional polyethers. produced by the Intergovernmental Oceanographic Commission (IOC) of UNESCO (United Nations Educational.g. Scientific and Cultural Organisation) has been updated (7). and is often necessary in light of the complex toxin profiles that can occur in shellfish tissue. Although most of these antibody-based test kits have yet to receive international regulatory approval. Another important development is the introduction of sensitive and selective antibody-based detection methods for a number of toxins.D. and a second updated edition dealing with the pharmacology. physiology and detection of marine and freshwater toxins has also appeared (8). In the last decade seafood monitoring has intensified and detection methods have continued to develop in selectivity and sensitivity. FCIC UNC Wilmington Center for Marine Science Marvin Moss Lane Wilmington NC 28409 United States of America 7. FISH AND SHELLFISH TOXINS Jeffrey L.
some are the result of biotransformation processes by the filter-feeding host. temperate. can become contaminated with these toxins. PSPs are watersoluble and heat stable so cooking has little or no effect on PSP toxicity. and another. with intoxications occurring in many countries even today (15). The Ncarbamoyl sulfates are readily hydrolysed to STX. 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. 142 . Around twenty PSP toxins are known. and a tropical species Pyrodinium bahamense (16). An account of this meeting detailing recommendations and scientific advice on marine biotoxins has been published (10). all of which contain a tetrahydropurine skeleton (Figure 7. Saxitoxin (STX) and neosaxitoxin (neoSTX) are the parent members of the group and are the most toxic. even crustaceans such as lobsters. 7. All PSPs degrade rapidly in alkaline conditions. and sub-tropical waters and habitats (15). neoSTX and Gonyautoxins (GTX) 1-4. and a review on the risk assessment of marine toxins has also been produced (14).2. Gymnodinium catenatum. The remarkable frequency of neurotoxins from marine dinoflagellates has been noted (13). Consequently.g. 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.FISH AND SEAFOOD has been produced (9). Many of these species are extremely common and are found throughout the world in a variety of cold.1). a great variety of shellfish from all latitudes. Saxitoxin Group instead of the Paralytic Shellfish Poisons). In addition. dealing with the detection methods and regulatory requirements for many marine toxins has been published (12). 7. A brief report on the status of rapid detection methods for phycotoxins in the European Union (EU) has appeared (11).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.1 Origins and distribution The PSP toxins are produced by several dinoflagellate species belonging to the genera Alexandrium. Not all the toxins found in shellfish extracts are microalgal products.
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. However.2.1. and fingers within 5 .30 minutes of consumption. 143 . Human symptoms begin with tingling and numbness of the lips. 7. tongue. Structure of saxitoxin and neosaxitoxin and their sulphated derivatives 7.FISH AND SHELLFISH TOXINS STX GTX2 GTX3 GTX5 (B1) C1 C2 NEO GTX1 GTX4 GTX6 (B2) C3 C4 R1 H H H H H H OH OH OH OH OH OH R2 H H OSO3 H H OSO3 H H OSO3 H H OSO3 - R3 H OSO3 H H OSO3 H H OSO3 H H OSO3 H R4 H H H SO3 SO3 SO3 H H H SO3 SO3 SO3 - Fig. leading to paralysis within a few hours and in extreme cases death by asphyxiation due to respiratory paralysis. this is never sufficient to render the seafood safe to consume. 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). the less potent C toxins are readily hydrolysed by acid to the more potent GTX toxins.2 Toxicology PSP toxins affect the central nervous system of mammals. Cooking does not destroy these compounds and although some of the toxins may be extracted into the cooking water. and humans seem to be particularly susceptible.
and more standards of individual PSP toxins are required to validate the complete profile to toxins that could occur in contaminated shellfish tissue. other derivatives have been reported. and a second rapid dipstick kit (MIST Alert™) is available for the qualitative detection of positive and negative samples (19). An antibody-based kit that can detect STX in shellfish is commercially available (RIDASCREEN®). 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. 24) (Figure 7. 17). While the chemical detection methods are most promising for the future. and while simple and relatively inexpensive to operate. namely oxidative conversion to fluorescent derivatives after High Performance Liquid Chromatography (HPLC) analysis (post-column derivatisation). a pre-column method has received Association of Analytical Communities (AOAC) approval (18).2. This kit was found effective in all the samples tested in Europe (20) and was applied recently in a survey of Scottish shellfish (21). and islands of the South Pacific. Since its discovery. After some development and various trials. as well as other parts. The mouse bioassay has been in place for decades and the method has been standardised. specifically 144 . 7. Alternative HPLC methods have subsequently reduced this to a single HPLC run. and has also been evaluated by a number of EU laboratories.2). TTX most commonly occurs in many species of puffer fish. Most intoxication events have occurred in South East Asia.8 µg/g) of shellfish or seafood tissue. Taiwan.3 Tetrodotoxin Tetrodotoxin (TTX) possesses a unique chemical structure and was the first marine toxin to be chemically characterised. However it remains as the monitoring method of choice in many countries.3. China.1 Origins and distribution Unlike many other marine toxins there is no temporal distribution of TTX episodes. or conversion before HPLC separation (pre-column derivatisation) and these have recently been thoroughly reviewed (12. a criticism has been that such methods are time consuming. 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. One approach has been to take advantage of the ready conversion of these toxins to fluorescent derivatives. Thailand. 23. 15. including Japan.FISH AND SEAFOOD 7. 7. There continues to be a significant effort applied to the development of alternative chemical and biochemical detection methods for these toxins.3 Detection methods The safe limit set by most countries for PSP toxins is 800 µg/kg (0. Two approaches have been taken here. and over ten analogs are known (22. Early post-column methods were somewhat time-consuming and required three separate HPLC runs to complete the analysis.
and blocks diffusion of sodium through the sodium channel. Structure of tetrodotoxin 7. this has expanded to include bacteria within the Alteromonas and Pseudomonas genera. In Japan. All of the observed toxicity is secondary to the action potential blockade.3 Detection methods TTX can be extracted from suspect tissue using dilute acid conditions. and even newts. Typically this has been accomplished using HPLC followed by post 145 . and once again the mouse bioassay has been used to detect the presence of the toxin. and ingestion of larger amounts leads to cardiovascular effects including bradycardia. the blue-ringed octopus. hypotension. 26). This was indeed confirmed and a number of marine Vibrios have been shown to be capable of TTX production (24. 7. Fig. nausea and vomiting. However TTX has been found to occur in a remarkable variety of genetically unrelated species including gastropods. Chemical methods for TTX detection have also been investigated and are mainly based on its smooth conversion by heating in an alkaline solution to a detectable fluorescent (C9) base.2 mg of purified toxin can be lethal.2. molluscs. preventing depolarisation and propagation of action potentials in nerve cells. The onset of symptoms usually occurs rapidly. 25. horseshoe crabs.3.FISH AND SHELLFISH TOXINS in the liver. 27). TTX acts on the central nervous system by binding extracellularly to site 1 of voltage-gated sodium channels. 7. Since then. where puffer fish of the genus Fugu are a delicacy.2 Toxicology Like the PSP toxins. and in mild cases results in distal muscle weakness. An oral dose of 1 . gonads and roe (24). 23.3. raising speculation that TTX is produced by a bacterial source. A review of TTX detection methods has been published (12. respiratory failure and coma. Severe poisoning causes paralysis and respiratory failure. and more recently by an actinomycete Norcardiopsis dassonvillei (26). it is served in speciality restaurants with qualified chefs to properly prepare the fish.
7. Structure of domoic acid and three naturally occurring photoisomers 7. Immunoassay methods have also been investigated. and while promising.3. both the toxin and the diatoms capable of producing it have been found to be distributed worldwide. but it is known that such congeners are less toxic than DA itself (30). but all of them are considerably less toxic than DA itself (29). 7. Surveys by a number of groups have identified over a dozen species of Pseudonitzschia capable of producing DA. Currently 12 isomers of DA have been reported (see Figure 7.4.3 for examples).1 Origins and distribution The cosmopolitan diatom Pseudonitzschia multiseries was identified as the culprit organism in the 1987 poisoning event (31).4 Amnesic Shellfish Poisoning (ASP) Toxins The parent compound of this group is domoic acid (DA). 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). Since the original episode of human poisoning following consumption of DA-contaminated shellfish in Canada in 1987 (28). a polar glutamate agonist. Fig. These isomers arise as a result of isomerisation and location of the double bonds in the side chain of the molecule. need further development (24). However this method has been criticised for being complex. but 146 .FISH AND SEAFOOD column oxidation to the fluorescent derivatives.
another diatom species Nitzschia navisvaringica. cormorants and pelicans (34). scallops. and this has led to ASP toxicity in birds. disorientation and coma. seizures.. but any impairment of the blood brain barrier results in additional risk. The original 1987 event was followed some years later by a more widespread episode along the western coast of the US and today toxin-producing strains of Pseudonitzschia have been found in the Gulf coast of the US and along the western shores of the US and Canada. and clams. 147 .4. has been reported as a medium-level producer of DA (33). 33). Pseudonitzschia australis. For example.FISH AND SHELLFISH TOXINS only three strains (P. and review of the toxicologic pathology of DA has been published (36). southern and western portions of Ireland. Recently. the Philippines. To a great extent. Indeed. Penetration of the blood brain barrier is poor in healthy subjects. and sea animals such as sea lions and even whales (35). 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. nausea. and Pseudonitzschia seriata) are considered to be high producers of DA (>10 pg/cell).2 Toxicology DA is an acidic amino acid and potent neurotoxin unaffected by cooking or steaming. Another factor is the movement of the toxin through the food chain. 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. The effects of DA may be augmented by the excitotoxic effects of glutamate and aspartate in the sample. and is mainly distributed in the blood. Four deaths were attributed to the Canadian incident and years later a few seriously affected individuals still display chronic neurological deficits. multiseries. the Mediterranean. as does impaired renal function which increases serum concentration and residence time of the toxin. 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. which has been well documented in studies along the west coast of the US. Okinawa and more northerly parts of Japan. herbivorous fish become toxic after ingesting strains of Pseudonitzschia spp. found in the Pacific around Vietnam.10% of the administered dose.39). The oral absorption of DA is around 5 . 7. including persistent short term memory deficits (36 . Levels of DA in the other strains are generally <1 pg/cell. and in locations around New Zealand (32. 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. Victims display a variety of symptoms including headache.
and this is typically the method of choice.. in Monterey Bay. Provided the concentrations in shellfish are high enough (>50 µg/g shellfish tissue). and dinophysistoxin-2 (DTX-2) (Figure 7.4). but just over 15 years ago a third toxin. California. although in the latter study the probes for Pseudonitzschia pungens.. DTX-2 was discovered in contaminated Irish shellfish (46). Since their first characterisation it was believed that the only causative agents of this group were OA and DTX-1 (45). are amenable to laboratory culture and consequently are used in many experiments involving studies of DSP toxins. However. 48. Alternatively. An EnzymeLinked ImmunoSorbent Assay (ELISA) kit (Biosense) for DA has been approved by the AOAC. The benthic Prorocentrum spp. A rapid immunodiagnostic kit is now available to detect DA in phytoplankton and is reported to work satisfactorily in this application in the field (43). produce a variety of other 148 . and an immunosensor method has also been reported (4). dinophysistoxin-1 (DTX-1).FISH AND SEAFOOD 7. though this method is most commonly used to analyse plankton and seawater (42). 49). the heterotrophic dinoflagellates Dinophysis spp. To further complicate matters. DTX-2 has been found to be extremely widespread throughout many European maritime countries and is often the major DSP toxin present. are the more potent source of toxins – often very low cells/ml of seawater can result in human poisoning. as can LC/MS. Detection levels can be further lowered by conversion of DA to the fluorescent fluorenylmethoxycarbonyl (FMOC) derivative.4.5 Diarrhetic Shellfish Poisoning (DSP) Toxins The active agents responsible for DSP incidents are a group of polyethers that include okadaic acid (OA). australis. In the context of this chapter. 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. Of additional interest is the use of molecular probes originally developed to detect the presence of cells of potentially toxic Pseudonitzschia spp. The probes have been employed in trial studies in New Zealand (44) and Scotland (43) to detect P. shellfish tissue can be efficiently extracted with aqueous methanol (1:1). the term “DSP toxins” will only be used to refer to these three polyether compounds and any derivative based on them. Pseudonitzschia fraudulenta and Pseudonitzschia delicatissima were found to be less effective. 7. DSP toxins are produced by dinoflagellates of the genera Prorocentrum and Dinophysis (47. 41). The sensitivity of the LC/UV (ultra violet) method (detection limit ~ 1 µg/g) can be improved (20 .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. and these methods have been reviewed (40. Dinophysis spp. and consequently were often overlooked as the real source of the toxins.30 ng/g) with a rapid clean-up step using a strong anion exchange resin. 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. Since then.
FISH AND SHELLFISH TOXINS lipophilic compounds that are toxic in the mouse bioassay. and Dinophysis norvegica among others (47. but do not have the same mechanism of action as DSP toxins nor induce the same symptoms. Dinophysis fortii. Today over ten Dinophysis spp. Prorocentrum maculosum. 7. they can be converted to the parent toxins by nonspecific esterases or lipases either in the producing organism or in the host shellfish. Dinophysis acuta. and DTX-6) all of which are rapidly converted to OA when the plankton cells are ruptured (48). This was probably not the first DSP episode. Fig. 49). Dinophysis caudata. Norway and Japan were likely due to DSP toxins produced by a Dinophysis spp. 149 . which include the pectenotoxins and the yessotoxins. 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. Lately these compounds. and anecdotal evidence now suggests that several seafood poisoning episodes during the 1960s and 1970s in the Netherlands. While these esters are not toxic in vitro. and include Dinophysis acuminata.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.5.4. such as Prorocentrum lima. and Prorocentrum belizeanum. DTX-5. In this case Dinophysis was suspected as the culprit organism. Structure of okadaic acid and the two related DSP toxins 7. 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. The parent acids are the only DSP congeners to be found in shellfish tissue. DSP toxins produced by benthic Prorocentrum spp. In comparison.. are also recognised as a threat to human health. analysis of Prorocentrum cultures have uncovered an additional series of diol esters of OA and sulphated diesters (called DTX-4. are recognised as DSP toxin producers.
vomiting.5. and DTX-2. various European countries. This regulatory level includes the possibility of LSPs in the DSP fraction being tested. DSP episodes have been reported worldwide and are included in many monitoring programs (47).FISH AND SEAFOOD Perhaps as a consequence of these multiple sources of DSP toxins. but the method is complex and matrix effects as well as interferences with other lipophilic compounds such as the pectenotoxins compromise the method.3 Detection methods Once again the mouse or rat bioassay is relied upon to detect DSP toxins in contaminated seafood.2 Toxicology The DSP toxins are potent inhibitors of the serine/threonine phosphatases PP1 and PP2A. some careful clean-up steps are required to avoid such matrix interferences and to simplify data interpretation (48). A regulatory level of 80 . 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).160 µg OA equivalents/kg of whole shellfish tissue has been implemented in some countries. 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.3 days. even with the minimal of clean up steps (48). although a comparative study of in vitro phosphatase inhibition with mouse bioassay data has shown that OA is about twice as potent as DTX-2 (51). and abdominal pain. 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. At the first Meeting of the EU National Reference Laboratories on Marine Biotoxins and Analytical Methods and Toxicity Criteria in 1996. which can lead to an underestimation of the total DSP content in a sample. All the DSP toxins have around a thousandfold greater affinity for PP2A than PP1.5. New Zealand and Chile. it was agreed that the established mouse bioassay (52) with an observation time of 24 hours is currently the preferred method for the detection of the acute toxicity of acetone soluble DSP toxins. DSP episodes have been reported from Japan. Since the late 1970s. nausea. it does not provide information on the DSP toxin profile. DSP-Check is a commercially 150 . 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. and a detection limit of 100 µg/kg has been reported (12). Human symptoms include diarrhoea. In recent years. and while this provides critical seafood safety information. The onset of symptoms can occur within 30 minutes of consuming contaminated shellfish and generally last about 2 . 7. Due to the lipophilic nature of the toxins. 7. Preparation of a fluorescent 9-anthryldiazomethane (ADAM) derivative followed by HPLC offers reasonable sensitivity. Immunoassays have been developed for OA detection but an inherent problem of these methods has been the low cross reactivity with DTX-1.
for many years these compounds were included in the DSP toxin complex.FISH AND SHELLFISH TOXINS available ELISA test kit that can detect both OA and DTX-1 with comparable sensitivity. Some doubt remains as to the significance of these compounds as marine or shellfish toxins with respect to human health since the PTXs for example are co-extracted with the DSP toxins. with good agreement with the mouse bioassay and LC–based methods (54. However. 55). Structure of the principal pectenotoxins and one of the seco-acid derivatives 151 . 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.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. Some representative examples of both groups of toxins are shown in Figures 7. Fig. they are toxic in the mouse bioassay and can result in false positives and hence are included here for completeness. Within this group.5. 7. with a claimed detection limit of 20 µg/kg. over 20 PTXs are known (56) and more than 30 YTXs have been reported (57. As a result these compounds are now being referred to as lipophilic shellfish toxins (LSTs) (49). 58). In yet another approach. and another rapid antibodybased test kit for DSP toxins has also been reported (53). fluorometric protein phosphatase inhibition assays have been found to perform better than colorimetric assays. 7. and the latter group are usually found to be responsible for any reported shellfish toxicity (59).5 and 7.6. If a European collaborative study of the fluorometric protein phosphatase inhibition method is successful it may eventually be approved for regulatory purposes in the EU.
many parts of Europe and the Mediterranean. 7. eastern Canada. The sources of YTXs have now been identified as the dinoflagellates Protoceratium reticulatum and Lingulodinium polyedrum. Structure of the two parent yessotoxins. 7. While 152 .6. 58).FISH AND SEAFOOD Fig.2 Toxicology PTXs show varied toxicity in the mouse bioassay (59). 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. YTX and homo-YTX 7. New Zealand and South America (49.1 Origins and distribution It is now well recognised that PTXs are co-produced with DSP toxins by various strains of the dinoflagellate Dinophysis and hence the distribution of both families of toxins in shellfish is similar (49). As is typical for many algal species. the toxin content and profile may vary among different strains. which may co-occur in blooms with other organisms.6. 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). Japan. 57. but nevertheless the toxins are extremely widespread and have been found in various shellfish in Norway and Scandinavia.
but less so following oral dosing (61). Many YTX toxins contain a conjugated diene system. The crude extract may be used directly in the mouse bioassay. For many years it was believed that 153 . but various congeners are acutely toxic in the mouse bioassay following IP injection. The regulatory level of 160 µg/kg shellfish tissue has been established and the mouse bioassay is often the primary detection method although LC/MS methods are preferred and this requires some simple clean-up of the crude extract prior to chemical analysis.FISH AND SHELLFISH TOXINS demonstrating considerable toxicity following IP injection. but a fluorescent derivative has been prepared for others. which is further compounded by differences in the experimental design. 58). the PTXs are susceptible to bioconversion in shellfish tissue and consequently it is often desirable to determine the profile of toxins present. While LC/UV detection is possible for PTXs. 64).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. Because of the number and complexity of the YTX group. 7. Acetone extracts of shellfish tissue are dried and re-suspended in 60% aqueous methanol before partitioning against dichloromethane. Estimates of acute toxicity of the parent compound following IP injection vary considerably. a regulatory level of 1 mg/kg shellfish tissue has been set. and this is believed to be due to hydrolysis of PTX-2 to the seco acid derivative. There are no clear cases of human illness resulting from ingestion of PTXs (59).7 Neurotoxin Shellfish Poisoning (NSP) Toxins NSP episodes are caused by a family of fused polyether compounds known as the brevetoxins (63. The YTXs are retained in the aqueous methanol fraction while the PTX and DSP toxins are concentrated in a dichloromethane fraction. There are two skeletal types known as PbTx-1 (or brevetoxin A) and PbTx-2 (brevetoxin B) (Figure 7. Although there is no evidence that yessotoxins are responsible for human illness. However. the PTXs show considerably less toxicity when administered orally.7). which aids detection by LC/UV. Some of the PTXs are potent cytotoxins in vitro (60). Their toxicology and mechanism of action remain uncertain. some of which are found as glycosides and sulfates. it may be compromised by other biological materials in the matrix.6. The EU has recommended an extraction and solvent partition process to separate the polar YTX compounds from the more lipophilic PTX and DSP compounds. The structures and chemical data including the molecular weight of each member have been collated (56). though its cell effects may be linked to modulation of calcium channels (62). 7. The YTX group has not been reported to cause human illness.
and Heretosigma akashiwo. Many of these species are associated with massive fish kills. Fibrocapsa japonica. and off the coast of New Zealand (65). They have also been associated with fish kills along the eastern mid-Atlantic coast of the US. Fig. whelks. and cockles (65. which are found in Japanese waters (66). verruculosa off the mid Atlantic coast of the U. the same toxins have been reported to be produced by three genera of Raphidophytes. oysters. namely Chatonella marina. 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).7. brevetoxins are found in mussels. 66).7. 154 . but in 1992/1993 they were identified as the culprits in a human poisoning event in New Zealand (65.S.FISH AND SEAFOOD NSP toxins only occurred in the Gulf of Mexico and along the western coast of Florida. In the Gulf of Mexico.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”. PbTx-1 and PbTx-2. 7. and one of the brevetoxin metabolites BTX-B2 7. 66). and recently the major toxin associated with an ichthyotoxic unialgal bloom of Chatonella cf. was chemically characterised as PbTx-2 (67). Interestingly.
The mean time to onset of symptoms is around 3 hours.7. A second generation competitive ELISA assay has been developed that has successfully quantified brevetoxins in a variety of biological and environmental matrices including seawater. Ingestion of brevetoxins results in a variety of symptoms including parathesia. malaise. Exposure to brevetoxins can also occur through inhalation of brevetoxin aerosols resulting in irritation of the upper respiratory tract (65. vertigo.7. beginning with herbivores that feed on benthic dinoflagellates associated with reefs and which are 155 . In this application. but this method is now used routinely in New Zealand and the US to monitor for the presence of brevetoxins (68). considerable progress has been made in the development of LC-MS methods to routinely monitor for brevetoxins in shellfish. with hot spots in the coral reef areas of the Caribbean and French Polynesia. Outbreaks of ciguatera poisoning are confined to tropical and sub-tropical regions of the ocean. The human health implications following exposure to NSPs have been reviewed (69). In a comprehensive multi laboratory evaluation of five different detection methods as possible replacements for the mouse bioassay. and it has also been used in studies to quantify occupational and recreational exposure to aerosolised brevetoxin exposure (73). and mammalian body fluids without pretreatment or dilution (72). The mouse bioassay is used as the primary detection method. shellfish homogenates. 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. which can result in a cascade of associated events in a cell (66).2 Toxicology Brevetoxins bind to site 5 of voltage-gated sodium channels. nausea.FISH AND SHELLFISH TOXINS although recently it has been shown that herbivorous fish can store brevetoxins in their flesh (68). and diarrhoea. 7. 69). 7. this family of toxins has proven to be a challenge both to identify and monitor. 7.8 Ciguatoxins Known for centuries and originally referred to as ciguatera by Spanish explorers in the Indo-Pacific ocean and Caribbean. and in severe cases seizure.3 Detection methods Three basic approaches for the detection of NSP toxins have been recently reviewed (71). 66. the competitive ELISA method correlated most closely with the mouse bioassay results. While the mouse bioassay continues to be used. pre-treatment or clean-up of the shellfish sample is essential in order to obtain reproducible results. with a mean duration of symptoms of around 1 day. The brevetoxin metabolites are reported to be less toxic than the parent compound (70). Ciguatera toxins (CTXs) are extremely potent and occur in finfish rather than shellfish.
8). 7. most likely a consequence of biotransformation of the parent toxins as they pass up the food chain. and other genera of neighbouring reef dinoflagellates can further contribute to an extremely complex profile of toxins in finfish. 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. Indeed even the chemistry of CTXs from the Pacific differs subtly from those found in the Caribbean (Figure 7. 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. and only a few structures have been identified. The diversity of benthic phytoplankton that exist on reefs further complicates the picture. Distribution through the food chain begins with 156 .FISH AND SEAFOOD transferred through the food chain to the larger carnivores. Culprit species producing CTXs may simultaneously produce unrelated toxins such as maitotoxin. Fig. but more are suspected to exist. lipid soluble polyethers. CTXs are large. a group of closely related polyethers that are considered the source of ciguatoxins. A detailed comprehensive review of ciguatoxins has recently been published (74). green and brown seaweeds and also occur in sediments and coral debris.8.8. complex. although the dinoflagellate Gambierdiscus toxicus is considered to be the major source of these toxins.
These include paresthesias (numbness and tingling of the extremities). myalgia. and inducing membrane depolarisation resulting in prolonged symptoms indicative of damage to nerve tissue. weeks or even months (74 . 78). Other symptoms include a metallic taste. Another symptom that can develop over time is extreme fatigue. sharks.2 Toxicology Ciguatoxins (and maitotoxin which is often co-produced with CTXs) are extremely poisonous.1 µg of ciguatoxin can result in human illness (74. Globally. and the sensation of loose teeth (77). and although the gastronomical problems may dissipate within a day or so. where cold objects feel hot and vice versa. though the mouse bioassay continues to be relied upon as the most common method of detecting CTXs. there are neither standards nor an official testing program.76). Even when suspect samples are available.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. arthralgia. they bind to voltage-dependant sodium channels in the cell membrane causing them to open. The pharmacological effects following CTX exposure are numerous and include severe gastrointestinal distress and neurological symptoms. and it is estimated that ingestion of as little as 0. all of which can be linked to the effect of CTX on excitable membranes. which in turn are consumed by carnivores such as snappers. although not all CTXs are equipotent. and moray eels. the absence of a strong chromophore means that simple detection methods based on UV or fluorescence-based detection methods are ineffective. 75). Like the brevetoxins. only trace amounts of CTXs can cause illness following consumption of contaminated fish. 7.8. A good knowledge and careful observation of the symptoms in the bioassay (Minimal 157 . 7.8. Local island populations attempt to avoid poisoning by catching only smaller fish.3 Detection methods As a consequence of their potency. 75. Differences in potency among the CTXs correlate directly with their binding affinity for the sodium channel. which can be confused with chronic fatigue syndrome. and temperature sensation reversal. barracudas. Patients have reported symptoms lasting weeks and even months. and in fact CTX poisoning is often only identified after consumption of contaminated fish. pruritus. the neurological symptoms can persist for days. but fish toxicity can vary widely depending on where the fish are caught and how long they have been feeding in that area. and there are suggestions that such incidents are increasing and spreading worldwide (74). Nevertheless CTX poisoning continues to be a threat to local and visiting populations. 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. The onset of symptoms can occur rapidly within 24 hours. groupers.
and following synthesis of the molecule some time later.30%) and AZA3 (5 . following consumption of contaminated mussels Mytelius edulis (82). almost 20 additional derivatives have been reported (83 . Recent reviews of this new group of toxins have appeared (83. and the corrected structure was published in 2004 (86) (Figure 7. which are based on an LC-MS/MS approach. The original structure proposed revealed an entirely new class of shellfish toxin. Recently.5) have been fully characterised by Nuclear Magnetic Resonance (NMR) and MS analysis.25 µg/kg) can provide useful clues as to the toxins present but more specific and informative assays are required.g. Immunoassays may be an alternative detection method (79).9 Azaspiracids Azaspiracid (AZA1) was originally identified in 1995 as the causative agent for a poisoning episode in Ireland.9). with AZA2 (10 . and another dealing with the chemistry and detection methods has been published (85). which features characteristic fragment ions corresponding to sequential losses of three molecules of water. Usually the additional AZA derivatives found in shellfish tissue are hydroxylated derivatives (e. only a few laboratories are able to undertake such analyses. and the separation of these polyethers from the lipid-soluble fraction of fish provides an additional challenge. although specificity and sensitivity remain major hurdles (74) and the presence of DSP toxins affects the test (80). at C-3 and C-23). and it is likely that most of these compounds are biotransformation products formed in the shellfish tissue. In this way Caribbean and Pacific CTX-1 compounds can be distinguished (74). high-throughput screen (74).FISH AND SEAFOOD Lethal Dose (MLD) 0. A sodium channel-specific assay using mouse neuroblastoma cells has been described although this approach has still to be developed in a routine. another in vitro sodium channel-based assay. has been developed.85). Usually the major AZA derivatives found in shellfish are AZA1 (50 .80%). Consequently. This is then subjected to LCMS/MS analysis. 84). Since the original discovery and identification of the toxin. The majority of these have been proposed based on LC/MS analytical data. 158 . Acetone extracts of fish tissue are subjected to solvent partitioning. and only a few structures (AZA 1 . 7. specific for CTX activity. Chemical detection methods are severely compromised by the lack of standards. but this only has application in a clinical setting (81).9). followed by clean-up on silica and aminopropyl cartridges.20%) present in lesser amounts (Figure 7. minor revisions of a few stereocenters were made.
). which can persist for 2 . but current data indicates that unlike the DSP compounds. and to date this has been the only source from which these toxins have been found at toxic levels. diarrhoea and stomach cramps. it has been impossible to establish that P. which are ingested by P. 90). and to date no long-term effects have been recorded (83. While this result is not necessarily in debate. oysters (Crassostrea gigas. it was reported that the heterotrophic dinoflagellate Protoperidinium cassipes was the planktonic source of the toxins (87). There has been considerable debate over the source of azaspiracids. 88.FISH AND SHELLFISH TOXINS Fig. AZA toxins do 159 .3 days. AZA occurrence has been reported by several European countries including France. The bivalves affected include mussels (Mytilus edulis. Portugal. clams (Tapes philipinarum. These symptoms are not unlike those experienced following the consumption of DSP toxins. However. 89). 84.1 Origins and distribution The azaspiracids were first isolated from contaminated shellfish. Donax spp.2 Toxicology Consumption of AZA contaminated shellfish can result in acute symptoms that include nausea. Following the original episode. scallops (Pecten maximus). cockles (Cerastoderma edule).9. 7. as in other cases of shellfish toxins. these molluscs were never considered to be the biogenetic source. Structure of the three principal azaspiracids found in shellfish 7.9. and some time after the original episode in Ireland. Mytilus galloprovincialis). and off the coast of Morocco (83. Ostrea edulis). 7.9. and most recently in a crustacean (Cancer pagurus). In fact the routes of trophic transfer of AZAs through the food chain ending in contaminated shellfish remain unknown. cassipes (83). vomiting. Scandinavia. Ensis siliqua. and variable field results have led to suggestions that the actual producing organisms are Dinophysis spp. cassipes produces AZAs in culture.
although autopsies conducted after 4 and 8 hours revealed various gastrointestinal (GI) abnormalities including accumulation of fluid in the ileum. The mouse bioassay has been used to monitor shellfish extracts. though the possibility exists that they may inhibit other phosphatases. and closely related ptierotoxins (Figures 7. 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). Due to their relatively recent discovery and limited knowledge of their toxicological properties and impact on human 160 . 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. 7. Using the same approach. This common cyclic imine feature is believed to be a common pharmacophore of this group of toxins which comprises the gymnodimines. abnormalities in the thymus and spleen. the spirolides. Based on IP minimum lethal dose data. but all contain a spirocyclic or cyclic imine function within their structure. which are particularly amenable to detection by positive ion electrospray mass spectrometry. 7. However.3 Detection methods The regulatory level for AZAs in shellfish has been set by the EU at 160 µg AZA equivalents/kg shellfish tissue. 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. and ASP. and erosion and bleeding in the stomach. 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. 91. Once again. Currently LC/MS and LC-MS/MS analysis is the preferred chemical analytical detection method for AZAs. AZAs also display cytotoxicity towards various mammalian cell lines (83. 85).FISH AND SEAFOOD not inhibit the phosphatases PP1 and PP2. PSP. 92). it is suggested that the order of toxicity is AZA2 (110 µg/kg) >AZA3 (140 µg/kg) >AZA (150 µg/kg). Importantly these pathological outcomes were reported as being different from those caused by DSP. The limited amounts of AZA toxins available continue to hinder a complete understanding of their mechanism of action. it is believed that the hydroxylated derivatives AZA4 (470 mg/kg) and AZA5 (<1000 mg/kg) are considerably less toxic. the pinnatoxins.9.12).10 Cyclic imines These toxins are represented by several distinct structural types. parenchymal cell pyknosis in the pancreas.10 – 7. Injection of a lethal dose of AZA1 in mice (>150 µg/kg) caused vacuole formation and fatty acid accumulation in hepatocytes. and necrosis of epithelial cells on the microvilli (91). Key fragment ions are useful in identifying the various AZA compounds that can occur in shellfish tissue (83. but unlike DSP monitoring methods which use only the hepatopancreas of the shellfish for extraction.
scallops.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.10.520 Da (94.10. two additional congeners. cockles. In only two cases (gymnodimines and spirolides) has a planktonic source been identified. Fig.. Some time later. Pacific oyster. gymnodimine B and C were isolated from the dinoflagellate Karenia selliformis (formerly Gymnodinium selliformis).10). The structure is novel in that it contains a spirocylic imine group and a single tetrahydrofuran ring embedded within a macrocyclic structure. 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. Reviews of the current status of the cyclic imine group were published by Molgo and Cembella (94. 95). At the time of the initial observation of gymnodimine. Subsequent taxonomic studies revised this again to a new dinoflagellate K. it was proposed that a dinoflagellate identified as Gymnodinium spp. 161 . 7. 7. they are sometimes referred to as emerging toxins of uncertain human health concern.FISH AND SHELLFISH TOXINS health. 95). surf clam and New Zealand abalone.10.1 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. 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 . but they are included here for completeness. selliformis (97). Structure of the cyclic imine gymnodimine 7. led to the identification of a new marine toxin with an unprecedented structure named gymnodimine (96) (Figure 7.1.
11). The original observation of toxicity in New Zealand followed from IP injection of lipophilic extracts in the mouse bioassay. Whether isolated from dinoflagellate cells directly or from shellfish tissue. and so spectroscopic detection by UV methods is not favoured. all or nothing” mouse bioassay response to gymnodimines and cyclic imines in general continues to be a useful monitoring approach.3 Detection methods The characteristic “fast-acting.1.100). 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. They were originally discovered in extracts of mussels (M. Fractions are then dried and re-suspended in methanol or acetonitrile in preparation for LC/MS analysis (94. Based on this diminished oral toxicity.1. 95) (Figure 7. The structure of the spirolides bears a strong resemblance to both gymnodimine and 162 .10.FISH AND SEAFOOD 7. However. There is a rapid onset of symptoms including respiratory distress. The time lapse between ingestion and death is between 5 and 15 minutes. Most cyclic imines lack an extended chromophore. cyclic imines are readily observed by electrospray mass spectrometry in the positive ion mode. finishing with 100% methanol. together with a lack of evidence of human intoxication. This led to the suggestion that gymnodimine acts by blocking nicotinic acetylcholine receptors at the neuromuscular junction. edulis) and scallops (Placopectin magellanicus) from eastern Canada that displayed abnormal symptoms in the mouse bioassay. abdominal breathing.10. Reduction of the imine group eradicated the toxicity.2 Toxicology A review of the toxicology of all cyclic imines has been published (98). In particular. extracts exhibited the fast acting symptoms that became associated with cyclic imines following IP injection. 50%) of 96 µg/kg. and there was a further ten-fold reduction in toxicity when the toxin was supplied with food. paralysis of the hind quarters and eventually complete immobility. and this is the detection method of choice.2 Spirolides The spirolides comprise the largest group of cyclic imine compounds.to10-fold reduction of toxicity (LD50 600 . it has been stated that regulation against gymnodimine is not required (98). and lack of responsiveness to stimuli. almost a dozen derivatives in total have been identified to date (94. 95). Since their original discovery which resulted in the characterisation of spirolides A-F (99.10. Oral administration of the toxin to mice by gavage resulted in about an 8. A later study using purified toxin established an LD50 dose (Lethal Dose.900 µg/kg). and short-acting cholinesterase inhibitors such as neostigmine or physostigmine protected mice against the toxic effects of gymnodimine. 7. 7.
While oral toxicity is considerably less than by IP injection. southern Ireland.1 Origins and distribution At the time of the original discovery of the spirolides in shellfish from the coastal waters of eastern Nova Scotia. or by oral ingestion (381 . these are the toxins that are also most resistant to acid hydrolysis of the imine function 163 . Fig. They have also been reported in the southern hemisphere in Chilean mussels and in New Zealand shellfish and plankton (94. those spirolides possessing the vicinal dimethyl grouping (see Figure 7. 7. the gulf of Maine. The LD50 values vary considerably depending upon whether the compound is administered by IP injection (5 . 7. All data reported here are for desmethyl spirolide C. in fjords in Denmark and Norway.2. Since then the spirolides have been found to be globally distributed and are found in shellfish or plankton fractions along eastern Canada. fasted mice). Structural differences among the various spirolides result from variations in the number of methyl groups and the sequence of spiroketal rings 6-5-5 in one group and 6-6-5 in the other. the western coast of Scotland.707 µg/kg.2 Toxicology The spirolides are the most studied group of the cyclic imines.11. the Bay of Biscay and most recently from Mediterranean shellfish and plankton in the Adriatic (102). Structure of four spirolide toxins A-D 7.10.FISH AND SHELLFISH TOXINS the pinnatoxins (vide infra). 95). Interestingly. R1 = CH3) of the cyclic imine ring are the most toxic of the group (98).2.8 µg/kg). This proved to be the case and eventually the dinoflagellate Alexandrium ostenfeldii was identified as the culprit organism (101). their localisation in the digestive gland of mussels and scallops led to the suspicion that plankton were the real source of the toxins. by gavage (87 .166 µg/kg.11. fasted mice).10.
7.FISH AND SEAFOOD (100). 95. consistent with modulation of acetyl cholinergic receptors (103). To date. 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. and in fact these compounds share a number of other structural features.12. Once again this relies on the sensitivity of these cyclic imines to detection by positive ion electrospray mass spectrometry.10. which includes two diastereomers.3 Detection methods As discussed for gymnodimine. 7. Structure of the pinnatoxin group and the closely related pteriatoxins 164 . Three members of this group have been reported.2.3 Pinnatoxins and pteriatoxins Pinnatoxins A . While the mechanism of action remains to be clearly defined. the mouse bioassay can be used to detect these fast acting toxins.12).10. However. 7. 102). Fig. The compounds are zwitterionic and therefore somewhat more polar than the spirolides. A number of methods following this procedure have been described (94. The pteriatoxins were isolated from a different mollusc but bear a strong structural resemblance to the pinnatoxins (Figure 7.B contain a spiro imine moiety similar to that found in the spirolides. four members of the group have been identified – two of them being the C-34 diastereomers. experiments have shown upregulation of muscarinic and nicotinic receptors in mice following dosing with spirolide C.
3. Only the acute toxicity in mice following IP injection has been reported.10. but the mouse bioassay data and LC-MS can be used to monitor suspect seafood for these compounds. there is no compelling evidence to suggest that they have resulted in human toxicity (98). once again reflecting improvements in purification and spectroscopic methods permitting structural identification of new and complex structures. 7. and later the closely related pteriatoxins were isolated from the digestive glands of another Okinawan bivalve Pteria penguin (105). This is generally a result of increased monitoring and continually improving detection technologies and methods. There is strong circumstantial evidence to suspect that these toxins. and more advanced means of detection.2 Toxicology There is little information describing the toxicology of these compounds (98). has come the realisation that a “toxin profile” can be a complex mixture of many different classes of toxins. This prompted a chemical investigation of Pinna muricata that ultimately led to the identification of the pinnatoxins in 1995 (104).10. Coupled with this. Despite this progress. 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. 99%) values for pinnatoxins A-C are in the range (22 .1 Origins and distribution Shellfish of the genus Pinna. to date action or 165 . 7. The LD99 dose for the pteriatoxins is reported to be in the range (8 . 7.3. have long been suspected of causing human illness after ingestion.11 Conclusion There has been considerable progress in the detection of known marine toxins.180 µg/kg).10.100 µg/kg).3. new toxin groups such as the azaspiracids and cyclic imines have also been reported. With increasing knowledge of the chemistry of toxins. which in many cases are now found to be more widespread than previously believed. which occupy tropical waters around Okinawa and the south east coast of China. while pinnatoxin D was essentially inactive. However. arise from an algal source . The LD99 (Lethal Dose.3 Detection methods No formal detection methods for these compounds have been reported. like the gymnodimines and spirolides.most likely a dinoflagellate. While this is recognised as a real possibility for a wide range of toxins. but this remains to be conclusively established (106).FISH AND SHELLFISH TOXINS 7. and in some cases it is known that one toxin can potentiate or augment the toxicity of the other.
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J.C. Henry M. Ed. 2004. 2002.. Cheng Y.A.. Buonocore M.. Ciguatera fish poisoning: treatment prevention and management.M.. in FAO Food and Nutrition Paper 80..M.S. Reich A.. Baden D. Johnson D. Baden D.S. Weisman R. 171 .. Baden D. Kirkpatrick B. Schrank K. Kubanek J. Bienfang P. Fernandez M. 75. Lenzi A. Chest. Pierce R.M.. 110.D. Intrinsic potency for synthetically prepared brevetoxin cysteine metabolites BTX-B2 and desoxy-BTX-B2. Baden D.D. Mild ciguatera poisoning: case reports with neurophysiological evaluations. Wanner A.. Granholm A. Abbott J.. Backer L.. Abraham W. Naar J. Jacocks H... Toxicon. Reich A. Kubanek J..FISH AND SHELLFISH TOXINS 65. 707-23. Clark R. Wang Z.C. Fleming L. 2007. Bossart G. Detection of brevetoxins in the twenty-first century. 69. 220-6. Poli M. 2002. 179-85.. Abraham W. Flewelling L. Marine Drugs.A. 110. Landsberg J. 23.. Whitney P. are potent ichthyotoxic neurotoxins that accumulate in fish... Ed. toxicology. Bean J. CRC Press...E. Flewelling L. Naar J. Gandini C. 2008. Boca Raton. Kirkpatrick B. 1598-603. Literature review of Florida red tide: implications for human health. 117-71...E.G. in Seafood and Freshwater Toxins.. 72. Cheng Y.B... Pierce R. Food and Agriculture Organisation of the United Nations..G. Ramsdell J.. Poon-King C.. New fish-killing alga in coastal Delaware produces neurotoxins. 825-34. Wolny J. 50.P. Ramsdell J.R. Environmental Health Perspectives. Toxicon.. 6. 479-516. Squicciarini D. Ed. 2008. 2004.. Competitive ELISA to detect brevetoxins from Karenia brevis (formerly Gymnodinium breve) in seawater.. 2008. Dickey R. 2008.J. and detection. 67. 456-79.H..M.. Marine Biotoxins. Food and Agriculture Organisation. 50. 131.S.. Naar J. Harmful Algae. 551-8.. Poon-King T. 71. Lancaster J.A. 519-50. CRC Press... Fleming L..G.. The molecular and integrative basis to brevetoxin toxicity... Zaias J. Butera R. Locatelli C. Backer L. 2000. Bourdelais A.. 74.N.P. 2007. shellfish and mammalian body fluid. Botana L. Fleming L. Environmental Health Perspectives.E.... 3. Aerosolised red tide toxins (Brevetoxins) and asthma. Boca Raton.. 2007. like ciguatoxins.E..M. Naar J. 465-70. West Indian Medical Journal. 70.. Granade R. Backer L.... Dechraoui M-Y.. Bottein M-Y.. Botana L.W. Boca Raton.E. Benson J.S. Neurologic Shellfish Poisoning (NSP). Food and Agriculture Organisation. 66.. 53..G.. 77. in Seafood and Freshwater Toxins.A.G. Steidinger K. Watkins S. Manzo L. Tomas C..... 2004. 76. Botana L. Freidman M. in Seafood and Freshwater Toxins. Ed. Ayyar R. Zaias J. Rome. Tomas C.. Ciguatera fish poisoning in industrial ship crew members: a retrospective study in a seaport general practice in Trinidad and Tobago. Baden D. 73..P. 68.. 99-115. Ciguatera toxins: Chemistry.. Bourdelais A. 187-94... Prockop L. Brevetoxins. Dickey R. Gannon D. Chen A. Muscle Nerve. CRC Press. Pierce R..
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.
hygiene managers. Establish a system to monitor control of the CCP.HACCP IN FISH AND SEAFOOD PRODUCT MANUFACTURE Step . Determine the CCPs. procedures.1 Assemble HACCP team HACCP requires management commitment of resources to the process. Verification . Establish the corrective action to be taken when monitoring shows that a CCP is not under control. Establish documentation concerning all procedures and records appropriate to these principles and their application. procedure. Each stage is discussed below in detail.3 Stages of a HACCP Study The HACCP system consists of the following seven basic principles: 1. 5. food microbiology. Validation . 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. 7.The application of methods.Obtaining evidence that the elements of the HACCP plan are effective. operation or stage in the food chain including raw materials. 6. An effective HACCP plan is best carried out as a multidisciplinary team exercise to ensure that the appropriate product-specific expertise is available. from primary production to final consumption. Figure 8. Conduct a hazard analysis. 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. 3. 2. food chemistry or engineering. quality assurance or control. 4. ingredient and packaging buyers. The team should also include 177 .A point. tests and other evaluations. 8.1 is a flow diagram illustrating this 12-stage logic sequence. 8. in addition to monitoring to determine compliance with the HACCP plan. Establish the critical limit(s).3. Establish procedures for verification to confirm that the HACCP system is working effectively.
g. If expert advice is not available on-site. chemical and/or physical). Logic sequence for application of HACCP 178 .FISH AND SEAFOOD personnel who are involved with the variability and limitations of the operations. Fig. biological. The scope of the plan should be determined by defining the extent of the production process to be considered and the categories of hazard to be addressed (e. it may be obtained from external sources. 8.1.
g. shelf life. marine toxins associated with particular fish and shellfish species.3.3. etc.g. The description should include information such as the product name. packaging.HACCP IN FISH AND SEAFOOD PRODUCT MANUFACTURE 8.3 Identify intended use The intended use should be based on the expected uses of the product by the enduser or consumer (e.). 8. However. or in aquaculture operations. it must be noted here that certain toxins (algal and shellfish) are not destroyed by heat treatments. is a cooking process required?). which should be described in detail. 8. heat treatment.3. pH. such as enteric viruses in shellfish.3. It should provide an accurate representation of all steps in the production process from raw materials to the end179 . aquaculture drug residues and potentially pathogenic Vibrio spp.).1 Fish and seafood The HACCP team should ideally have access to expertise on the appropriate fisheries. Expertise on such inherent hazards. fermentation. storage and distribution conditions and instructions for use. contaminated fish and shellfish should not be used.1.3.4 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. Vulnerable groups of the population. In addition. etc. the only CCP that can render the product safe from pathogenic organisms is adequate heat treatment during preparation. It is also important to identify the consumer target groups.2 Describe the product It is important to have a complete understanding of the product. freezing. naturally present on fish from warm waters (> 15 °C). 8. These factors are likely to have a significant effect on some potential hazards. 8. for example contamination by foodborne pathogens as a result of time/temperature abuse or unsatisfactory handling practice. including practices onboard fishing vessels. may need to be considered specifically. there may be species-specific hazards related to certain fish and shellfish. processing conditions (e. 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. physical and chemical structure (including water activity (aw). such as children or the elderly.3. composition.1 Fish and seafood Certain products may be contaminated or carry pathogenic organisms as a part of the natural flora.
time/temperature data.1 Fish and seafood Examples of flow diagrams for specific fish products may be found in the appropriate product chapters. For example. The operation should be observed at all stages. adequate chilling of finfish immediately after capture is vital for the control of several potential hazards.3. or circumstances of any reclaim or rework activity. Ideally. because the hazard analysis and decisions regarding CCPs are based on these data.6 List all potential hazards associated with each step. 180 ● ● . conduct a hazard analysis.3. It is also important to include observation of production outside normal working hours such as shift patterns and weekend working. survival or multiplication of pathogenic microorganisms. 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.3. 8. Many fish-processing operations have relatively few steps and the flow diagrams appear simple. features of equipment design.FISH AND SEAFOOD product. The analysis is likely to include consideration of: ● the likely occurrence of hazards and the severity of their adverse health effects. If HACCP studies are applied to proposed new process lines or products. it should also include details of CCP steps.4.5 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. It may include details of the factory and equipment layout. 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 qualitative and/or quantitative evaluation of the presence of hazards. then any pre-drawn HACCP plans must be reviewed once the lines/products are finalised. once determined. it is essential that the details of each step are fully appreciated and recorded. and therefore a detailed description of the procedure applied must be available. and any discrepancies between the flow diagram and normal practice must be recorded and the diagram amended accordingly. However. It is essential that the diagram is accurate. cleaning and hygiene procedures and storage conditions. ingredient specifications. 8.
Some hazards may require more than one control measure for adequate control and a single control measure may act to control more than one hazard. 8. The HACCP team should then determine what control measures exist that can be applied for each hazard. or desirable. where a hazard is repeated. Salmonella spp.g.1 Fish and seafood The term 'fish and seafood' includes an extremely varied group of products. and there are an equally varied range of potential hazards associated with them. or poor control of batter mixes in frozen battered fish portions may allow growth of Staphylococcus aureus and production of enterotoxin.6. One control measure may be relevant to several process steps. Pathogens may be part of the resident microflora of the living animal (e. modified-atmosphere packaging (MAP) of chilled raw fish may provide conditions suitable for the growth of psychrotrophic C. Vibrio spp. There may also be inherent hazards from parasites such as the roundworm Anisakis simplex and from marine toxins such as ciguatera in reef fish (derived from microalgae) and scombrotoxin (biogenic amines) development in fish containing high levels of histidine. and viruses). any food safety objectives or manufacturer’s food safety requirements. since this may interfere with the analysis. For example. the hurdle effect. there are particular hazards associated with contamination of shellfish by microorganisms from human sewage. Hazards specific to certain types of product are detailed in the appropriate chapters of this manual.HACCP IN FISH AND SEAFOOD PRODUCT MANUFACTURE ● ● ● ● production or persistence of toxins. For example. Note: it is important at this stage that no attempt is made to identify CCPs. Hazards introduced during processing of fish products depend very much on the characteristics of the process. botulinum. The incidence of psychrotrophic. or may originate from polluted water or from post-capture contamination (e. and the potential growth of Listeria monocytogenes on smoked fish.3. to generalise about expected hazards and the reader is once again referred to the appropriate product chapter for additional advice on specific hazards. 181 .g. Many of the microbiological hazards associated with fish and seafood products are derived from the raw materials. non-proteolytic Clostridium botulinum on fresh fish is also sufficiently high that its presence may be assumed. the number of consumers potentially exposed and their vulnerability. Therefore it is not possible.).
7. but for which no control exists at that step or any other. critical limits should be specified and validated for each CCP. 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 .2) to provide a logical. However. application of the decision tree should be flexible. given the enormous variety of fish products and processes in use. 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. biological or sensory parameters. it can be said that effective control measures are likely to include the following: • • • • • 8.3. chemical.1 Fish and seafood Specific product chapters provide information on criteria that may be used to set critical limits. Where possible.FISH AND SEAFOOD 8. structured approach to decision making.7 Determine CCPs The determination of CCPs in the HACCP system is facilitated by a decision tree (Figure 8.1 Fish and seafood Again. 8.3. and its use may not always be appropriate.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.8. Criteria used to set critical limits must be measurable and may include physical. More than one critical limit may be defined for a single step. However. 8. it is not possible to generalise on likely CCPs.3. For example.3. If a significant hazard has been identified at a step where control is required for safety. It is also essential that the HACCP team has access to sufficient technical data to determine the CCPs effectively. it is necessary to specify both time and temperature for a thermal process. and the reader is referred to the appropriate product chapter. then the process must be modified to include a control measure.
HACCP IN FISH AND SEAFOOD PRODUCT MANUFACTURE
• • • 8.3.9
Water activity (aw) values for dried fish pH values in fermented fish Microbiological quality of water in shellfish production areas Establish a monitoring system for each CCP
Monitoring involves planned measurement or observation of a CCP relative to its critical limits. Monitoring procedures for target levels must be able to detect loss of control of the CCP, and should provide this information with sufficient speed to allow adjustments to be made to the control of the process before the critical limits are violated. Monitoring at critical limits should be able to rapidly detect when the critical limit has been exceeded. Monitoring should therefore either be continuous, or carried out sufficiently frequently to ensure control at the CCP. Therefore, physical and chemical on-line measurements are usually preferred to lengthy microbiological testing. However, certain rapid methods, such as ATP assay by bioluminescence, may be useful for assessment of adequate cleaning, which could be a critical limit for some CCPs, for example pre-start-up hygiene. Persons engaged in monitoring activities must have sufficient knowledge, training and authority to act effectively on the basis of the data collected. These data should also be properly recorded. 8.3.10 Establish corrective actions
For each CCP in the HACCP plan, there must be specified corrective actions to be applied if the CCP is not under control. If monitoring indicates a deviation from the critical limit for a CCP, action must be taken that will bring it back under control. Actions taken should also include proper isolation of the affected product, repair of defective equipment, and an investigation into why the deviation occurred. A further set of corrective actions should relate to the target levels, if process drift is occurring. In this case, only repair of the process defect and investigation of the fault are required. All corrective actions should be properly recorded. 8.3.11 Establish verification procedures
Verification usually involves auditing and testing procedures. Auditing methods, procedures and tests should be used frequently enough to determine whether the HACCP system is being followed, and is effective at controlling the hazards. These may include random sampling and analysis, including microbiological testing. Although microbiological analysis is generally too slow for monitoring purposes, it can be of great value in verification, since many of the identified hazards are likely to be microbiological. In addition, reviews of HACCP records are important for verification purposes. These should confirm that CCPs are under control and should indicate the nature
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of any deviations and the actions that were taken in each case. It is also useful to review customer returns and complaints regularly. 8.3.12 Establish documentation and record keeping
Efficient and accurate record keeping is an essential element of a HACCP system. The procedures in the HACCP system should be documented. ● ● Determination of critical limits The completed HACCP plan
Examples of documented procedures include: ● ● The hazard analysis Determination of CCPs
Examples of recorded data include: ● ● ● Results of monitoring procedures Deviations from critical limits and corrective actions Records of certain verification activities, e.g. observations of monitoring activities, and calibration of equipment.
The degree of documentation required will depend partly on the size and complexity of the operation, but it is unlikely to be possible to demonstrate that an effective HACCP system is present without adequate documentation and records. The length of time that records are kept will be as per company policy, but should not be less than one year beyond the shelf life of the product. Three to five years is typical for many food companies. 8.4 Implementation and review of the HACCP plan
The completed plan can only be implemented successfully with the full support and co-operation of management and the workforce. Adequate training is essential and the responsibilities and tasks of the operating personnel at each CCP must be clearly defined.
HACCP IN FISH AND SEAFOOD PRODUCT MANUFACTURE
Fig. 8.2. CCP Decision Tree (Adapted from Codex Alimentarius Commission, 2003) Finally, it is essential that the HACCP plan be reviewed following any changes to the process, including changes to raw materials, processing conditions or equipment, packaging, cleaning procedures and any other factor that may have an effect on product safety. Even a small alteration to the product or process may invalidate the HACCP plan and introduce potential hazards. Therefore, the implications of any changes to the overall HACCP system must be fully
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considered and documented, and adjustments made to the procedures as necessary. Triggered reviews/audits should occur as a result of changes, whereas a scheduled review/audit should be annual, as a minimum. 8.5 References
Wareing P.W., Carnell A.C. HACCP – A Toolkit for Implementation. Leatherhead, Leatherhead Food International. 2007. Bernard D., Scott V. Hazard Analysis and Critical Control Point System: use in controlling microbiological hazards, in Food Microbiology: Fundamentals and Frontiers. Eds. Doyle M.P., Beuchat L.R. Washington D.C., ASM Press. 2007, 971-86. Su Y-C., Daeschel M.A. Sanitation and HACCP, in Surimi and Surimi Seafood. Ed. Park J.W. Boca Raton, CRC Press. 2005, 141-62. Huss H.H. Considerations in the application of the HACCP principles to seafood production, in Assessment and Management of Seafood Safety and Quality. Eds. Food and Agricultural Organisation, Huss H.H., Ababouch L., Gram L. Rome, FAO. 2004, 153-77. Chen H-C., Chang P.C-M. Frozen seafood safety and HACCP, in Handbook of Frozen Foods. Ed. Hui Y.H., Cornillon P., Legaretta I.G., Lim M.H., Murrell K.D., Nip WK. New York, Marcel Dekker. 2004, 339-63. Mortimore S., Mayes T. The effective implementation of HACCP systems in food processing, in Foodborne Pathogens: Hazards, Risk Analysis and Control. Eds. Blackburn C.deW., McClure P.J. Cambridge, Woodhead Publishing Ltd. 2002, 229-56. Mayes T., Mortimore C.A. Making the Most of HACCP: Learning from Other’s Experience. Cambridge, Woodhead Publishing. 2001. Mortimore S.E., Wallace C., Cassianos C. HACCP (Executive Briefing). London, Blackwell Science Ltd. 2001. Chartered Institute of Environmental Health. HACCP in Practice. London, Chadwick House Group Ltd. 2000. Jouve J.L. Good manufacturing practice, HACCP, and quality systems, in The Microbiological Safety and Quality of Food, Volume 2. Eds. Lund B.M., BairdParker T.C., Gould G.W. Gaithersburg, Aspen Publishers. 2000, 1627-55. Stevenson K.E., Bernard D.T. HACCP: A Systematic Approach to Food Safety. Washington D.C., Food Processors Institute. 1999. National Seafood HACCP Alliance. IQF cooked shrimp processing HACCP model, in HACCP User's Manual. Ed. Corlett D.A. Gaithersburg, Aspen Publishers. 1998, 454-73. Mortimore S., Wallace C. HACCP: A Practical Approach. Gaithersburg, Aspen Publishers. 1998.
HACCP IN FISH AND SEAFOOD PRODUCT MANUFACTURE Khandke S.S., Mayes T. HACCP implementation: a practical guide to the implementation of the HACCP plan. Food Control, 1998, 9 (2-3), 103-9. Forsythe S.J., Hayes P.R. Food Hygiene, Microbiology and HACCP. Gaithersburg. Aspen Publishers. 1998. Food and Agriculture Organisation. Food Quality and Safety Systems: A Training Manual on Food Hygiene and the Hazard Analysis and Critical Control Point (HACCP) System. Rome, FAO. 1998. US Food and Drug Administration. Fish and Fishery Products Hazards and Controls Guide. Rockville, Food Drug Administration. 1998. National Advisory Committee on Microbiological Criteria for Foods. Hazard Analysis and Critical Control Point Principles and Application Guidelines. 1997. Gardner L.A. Testing to fulfil HACCP (Hazard Analysis Critical Control Points) requirements: principles and examples. Journal of Dairy Science, 1997, 80 (12), 3453-7. Martin R.E., Collette R.L, Slavin J.W. Fish Inspection, Quality Control and HACCP: A Global Focus. Proceedings of a Conference, Arlington, May 1996. Lancaster P.A., Technomic Publishers. 1997. Codex Alimentarius Commission. Hazard Analysis Critical Control Point (HACCP) System and guidelines for its application, in Food Hygiene: Basic Texts. Ed. Codex Alimentarius Commission. Rome, FAO. 2003, 21-31. Garrett E.S., Jahncke M.l., Tennyson J.M. Microbiological hazards and emerging food safety issues associated with seafoods. Journal of Food Protection, 1997, 60 (11), 1409-15. National Seafood HACCP Alliance. HACCP: Hazard Analysis and Critical Control Point Training Curriculum. Raleigh, North Carolina State University. 1997. Dillon M., McEachern V. HACCP and quality assurance of seafood, in Fish Processing Technology. Ed. Hall G.M. London, Blackie. 1997, 224-48. Savage R.A. Hazard Analysis Critical Control Point: a review. Food Reviews International, 1995, 11 (4), 575-95. Pearson A.M., Dutson T.R. HACCP in Meat, Poultry and Fish Processing. Glasgow, Blackie. 1995. Huss H.H., Food and Agriculture Organisation. Assurance of Seafood Quality. FAO Fisheries Technical Paper No. 334. Rome, FAO. 1994. Bonnell A.D. Process control, in Quality Assurance in Seafood Processing: A Practical Guide. Ed. Bonnell A.D. London, Chapman & Hall. 1994, 39-53. Pierson M.D., Corlett D.A., Institute of Food Technologists. HACCP: Principles and Applications. New York, Van Nostrand Reinhold. 1992. Hudak-Roos M., Garrett E.S. Monitoring, corrective actions and record keeping in HACCP, in Quality Assurance in the Fish Industry: Proceedings of an International Conference, Copenhagen, August 1991. Eds. Huss H.H., Jakobsen M., Liston J. Amsterdam, Elsevier. 1992, 521-31.
Volume 4: Application of the Hazard Analysis Critical Control Point (HACCP) System to Ensure Microbiological Safety and Quality. Bryan F. Simple users' guide to the hazard analysis critical control point concept for the control of food microbiological safety. Geneva. Hazard Analysis Critical Control Point Evaluations: A Guide to Identifying Hazards and Assessing Risks Associated with Food Preparation and Storage. 33-44. WHO.HACCP IN FISH AND SEAFOOD PRODUCT MANUFACTURE Huss H.H. 3 (1). 14-9. Food Control. International Journal of Food Microbiology. 1992. 188 .. Mayes T. Blackwell Scientific Publications. 15 (1/2). 1992. World Health Organisation. 1992. 1988. Microorganisms in Foods. International Commission on Microbiological Specifications for Foods.L. Development and use of the HACCP concept in fish processing. Oxford.
The new legislation was designed to establish conditions. 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. under which food is produced to optimise public health and to prevent. transport and delivery vehicles requirements. to personnel requirements. have been replaced by several linked measures on food safety rules and associated animal health controls. including microbiological criteria. Hygiene is a general concept that covers a wide subject area. with the intention of introducing consistency and clarity throughout the food production chain from primary production. however. The general food hygiene Directive 93/43/EEC and other Directives on the hygiene of foodstuffs and the health conditions for the production and placing on the market of certain products of animal origin intended for human consumption. A number of standards are provided in food legislation. and conditions of raw materials. eliminate or acceptably control pathogen contamination of food.9. and one that plays an important role in most countries’ food legislation. from structural conditions in the factory or process facility. final product specifications. the existence of microbiological standards alone cannot protect consumer health. A new package of European Commission (EC) hygiene measures became applicable on 1 January 2006 to update and consolidate the earlier 17 hygiene directives. to sale or supply to the final consumer.1 Introduction Hygiene is an important aspect of ensuring food safety. Jenny Pfleger Leatherhead Food International Randalls Road Leatherhead Surrey KT22 7RY United Kingdom 9. Procedures under the new legislation are based on risk assessment and management and follow a 'farm to fork' approach to food 189 . EC FOOD HYGIENE LEGISLATION Eugenia Choi and Dr. Microbiological standards have a useful role and help establish requirements for the microbiological safety and quality of food and raw materials.
9. Prescribed are detailed measures to ensure the safety and wholesomeness of food during preparation. and sets out what those enforcing the provisions have to do. p. A number of more detailed implementing and transitional measures have been adopted at EC level. Subsequently.B. Regulation 854/2004 relates to the organisation of official controls on products of animal origin.1). the following European Union (EU) hygiene regulations have applied: • • • Regulation (EC) No.1992. 41). 7. p. 852/2004 of the European Parliament and of the Council on the hygiene of foodstuffs Regulation (EC) No. N. distribution. 2073/2005 on microbiological criteria for foodstuffs • The general hygiene requirements for all food business operators are laid down in Regulation 852/2004. 24.7. processing.15). 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. transportation.FISH AND SEAFOOD safety with the inclusion of primary production in food hygiene legislation.1991. Council Directive 91/493/EEC of 22 July 1991 laying down the health conditions for the production and the placing on the market of fishery products (OJ L 268. p. Regulation 853/2004 supplements Regulation 852/2004 in that it lays down specific requirements for food businesses dealing with foods of animal origin. European Communities L 268. 24.1991. packaging. manufacturing. storing. handling and offering for sale or supply to the consumer.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. as last amended by Regulation (EC) No 806/2003.2 Legislative Structure From 1 January 2006. 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. 9. J. 190 • • . 853/2004 of the European Parliament and of the Council laying down specific hygiene rules for food of animal origin Regulation (EC) No. as last amended by Regulation (EC) No 806/2003.
and also if the producer supplies small amounts of primary product to the final consumer or local retail establishments supplying the final consumer. the legislation is directly applicable law. and binding in its entirety on all member states from the date of entry into force. e. and of hunting and fishing. 1 . setting offences. 2006 No. Although the regulations have the force of law.1993.) in England. of stock farming. processing and distribution of food under their control satisfy the relevant hygiene requirements laid down in Regulation (EC) No. 19. 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. etc.I. 2073/2005.7. Wales and Northern Ireland. 852/2004 on the General Hygiene of Foodstuffs Food business operators must ensure that all stages of production. 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. 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. 9. right through to the implementation of procedures based on HACCP principles. domestic preparation or handling. 852/2004. particularly taking into account a number of factors ranging from ensuring food safety throughout the food chain to begin with primary production. revocation of existing implementing legislation. There are some exemptions. is required to give effect to the EU regulations. As regulations. as amended) came into force on 11 January 2006 (separate but similar national legislation also came into force on that day in Scotland. Primary products are defined as products of primary production including products of the soil. Additionally the requirements of EC Regulation 853/2004 must be complied with.14. Wales and Northern Ireland).I. p. national legislation in the form of a Statutory Instrument (S. food storage that is for private or domestic consumption.g.11) The EU hygiene regulations apply to all stages of food production including primary production. penalties and powers of entry. 191 . and equivalent legislation in Scotland.3 Regulation (EC) No.LEGISLATION • Council Directive 93/43/EEC of 14 June 1993 on the hygiene of foodstuffs (OJ L 175. with primary production. The Food Hygiene (England) Regulations 2006 (S. The national legislation in all four UK countries also applied the provisions of the EU Microbiological Criteria Regulation No. This Regulation lays down general rules for food business operators on the hygiene of foodstuffs. which will be covered later in this chapter. for example.
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Annex I – Primary production
Annex I (Part A) relates to general hygiene provisions for primary production and associated operations covering: (a) the transport, storage and handling of primary products at the place of production, (b) the transport of live animals, (c) for fishery products - transport operations to deliver primary products (which have not been substantially altered) from the place of production to an establishment Food business operators have the responsibility to ensure primary products are protected against contamination. Any community and national legislation relating to the control of hazards in primary production such as measures to control contamination resulting from surroundings e.g. air, soil, water etc., and measures relating to animal health and welfare, and plant health that may impact on human health should be complied with. Requirements for record keeping are also laid down. This relates to animal feed (nature and origin), veterinary medicines administered to animals (date given and withdrawal periods), any diseases, analysis of samples from other animals that might impact on human health as well as reports on animal checks performed. Part B of Annex I contains recommendations for guides to good hygiene practice. 9.3.2 Annex II – Stages other than primary production
Annex II of the regulation lays down additional general hygiene requirements that must be met by food business operators carrying out production, processing and distribution of food at all stages following primary production. A summary of Chapters I to IV of Annex II is provided as follows: 184.108.40.206 Chapter I Chapter I applies to all food premises, except premises to which Chapter III applies. Food premises must be kept clean and maintained in good repair and condition. Their layout should allow for this. The environment should allow good hygiene practices and provide temperature-controlled handling and storage conditions where necessary ,and enable foods to be kept at correct temperatures and be monitored.
Additionally, there are requirements for provision of adequate lavatories, basins, ventilation, lighting and drainage.
220.127.116.11 Chapter II Chapter II applies to all rooms where food is prepared, treated or processed, except dining areas and premises to which Chapter III applies. The design and layout of rooms should allow for good hygiene practices between and during operations. Therefore floor and wall surfaces, ceilings and windows should be constructed to prevent dirt accumulating. Surfaces where food is handled must be well-maintained and allow easy cleaning and disinfection, preferably using smooth, washable corrosionresistant and non-toxic materials. There should be facilities for cleaning or disinfecting, and for storing working utensils or equipment. Clean potable water and adequate provision for washing food is needed.
18.104.22.168 Chapter III Chapter III applies to temporary premises (e.g. marquees, market stalls, and mobile sales vehicles), premises used primarily as a private dwelling-house but where foods are regularly prepared for placing on the market, and vending machines. Premises and vending machines should be practically sited, designed, constructed and kept clean and maintained in good repair and condition so as to avoid the risk of contamination, in particular by animals and pests. Facilities should allow adequate personal hygiene, and surfaces in contact with food should be easy to clean. Enough potable water and storage arrangements for hazardous or inedible substances are required as well as adherence to food safety requirements.
22.214.171.124 Chapter IV Chapter IV applies to all transportation. Conveyances and/or containers used for transporting foodstuffs are to be kept clean and maintained in good repair and condition to protect foodstuffs from contamination and are, where necessary, to be designed and constructed to permit adequate cleaning and/or disinfection. Food should be maintained at appropriate temperatures.
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126.96.36.199 Chapter V Chapter V refers to equipment requirements. Adequate cleaning and disinfection is to be carried out frequently for articles, fittings and equipment in contact with food where contamination needs to be avoided. Equipment should be installed to allow adequate cleaning, and be fitted with the required control device.
188.8.131.52 Chapter VI Chapter VI refers to food waste. Food waste, non-edible by-products and other refuse is to be removed from rooms where food is present as quickly as possible to avoid accumulation. Such waste is to be deposited in closable containers, to allow easy cleaning. Refuse stores should allow easy cleaning and be free of pests. Waste must be eliminated hygienically in accordance with European Community legislation.
184.108.40.206 Chapter VII Chapter VII refers to the water supply. There must be an adequate supply of potable water, which should be used whenever necessary to ensure foodstuffs are not contaminated. There are also requirements for recycled water, ice in contact with food, use of steam and for the water used in the cooling process for heat-treated foods in hermetically sealed containers. Specific requirements are that clean water may be used with whole fishery products. Clean seawater, may be used for live bivalve molluscs, echinoderms, tunicates and marine gastropods; and for external washing. If it is used, there must be adequate facilities to ensure the supply does not create a source of contamination for the foodstuff. Recycled water used in processing or as an ingredient must not present a contamination risk. It must be of the same standard as potable water, unless the competent authority is satisfied that the quality of water cannot affect the wholesomeness of food in the finished form.
Additionally, ice that comes into contact with food or which could possibly contaminate it needs to be made from potable water or, in the case of chilling whole fishery products using clean water. Ice must be made, handled and stored under conditions that protect it from contamination.
220.127.116.11 Chapter VIII Chapter VIII sets out personal hygiene requirements for those working in a food handling area including clean protective clothing and the requirement that those carrying or suffering from a disease are not permitted to handle food. 18.104.22.168 Chapter IX Chapter IX covers provisions applicable to foodstuffs. A food business operator should not accept raw materials or ingredients, other than live animals, or any other material used in processing products, if they are known to be contaminated with parasites, pathogenic microorganisms or foreign substances, to be toxic or decomposed to such an extent that, even after the business operator applied normal hygienic processing , the product would be inedible. Raw materials must be kept under appropriate conditions throughout production, processing and distribution. In particular, temperature control (i.e. cold chain and food thawing) requirements are laid down.
22.214.171.124 Chapter X Chapter X lays down provisions applicable to the wrapping and packaging of foodstuffs to avoid contamination of any form. 126.96.36.199 Chapter XI Chapter XI lays down heat treatment requirements for food that is placed on the market in hermetically sealed containers. The main relevant process parameters (particularly temperature, pressure, sealing and microbiology) must be checked by the food business operator to ensure that the required heat treatment has been achieved. Also, the process used should comply with an internationally recognised standard (e.g. pasteurisation, ultra high temperature or sterilisation). 188.8.131.52 Chapter XII Chapter XII states training requirements for food business operators to ensure that food handlers are trained in food hygiene matters and in the application of HACCP principles.
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The Regulation requires that food business operators must notify the relevant competent authority of each establishment under their control that carries out any of the stages of production, processing and distribution of food, with a view to the registration of each establishment. Food business operators must also ensure that the competent authority always has up-to-date information on establishments, including the notification of significant changes in activity, and closure of an existing establishment. Food business operators must ensure that establishments are approved by the competent authority, following at least one on-site visit, when approval is required by the national law of the Member State in which the establishment is located, or under Regulation (EC) No. 853/2004, or by a separate decision adopted. Separate rules apply for businesses producing products of animal origin. 9.3.4 HACCP
Food business operators, other than at the level of primary production, and associated operations must put in place, implement and maintain a permanent procedure or procedures based on principles of the system of hazard analysis and critical control points (HACCP). Emphasis is placed on risk-related control, with responsibility placed on the proprietor of the food business to ensure that potential hazards are identified and systems are developed to control them. Under HACCP, food business operators must amongst other considerations, identify hazards to be prevented or eliminated or reduced to acceptable levels, identify critical control points (CCP) at which control is essential to prevent, eliminate or reduce hazards and establish critical limits at these points, implement effective monitoring procedures, and establish corrective actions in the case where a CCP is out of control. Procedures must be followed to confirm the above is in place and up-todate, as well as providing the competent authority with documents and records as evidence, when required. 9.4 Regulation (EC) No. 853/2004 Laying Down Specific Hygiene Rules for Food of Animal Origin
Regulation (EC) No. 853/2004 lays down hygiene rules for products of animal origin that apply in addition to the general hygiene rules of Regulation (EC) No. 852/2004. 9.4.1 Definitions
The following definitions apply for fishery products and live bivalve molluscs:
where appropriate. 197 . or in natural sites. if necessary: chilling or freezing: filleting. Purification centre means an establishment with tanks fed by clean seawater in which live bivalve molluscs are placed for the time necessary to reduce contamination to make them fit for human consumption. shucking. purification centres or dispatch centres in tanks or any other installation containing clean seawater. including products packaged under vacuum or in a modifiedatmosphere that have not undergone any treatment to ensure preservation other than chilling. heading. 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. where necessary. and chopping. gutting and removal of fins and. live echinoderms. skinning. to preserve or to improve organoleptic qualities and to ensure they are in a good state of vitality before wrapping or packing. reptiles and frogs) whether wild or farmed and including all edible forms. slime or sand. followed by wrapping or packaging. heading. whether whole or prepared. Factory vessel means any vessel on board which fishery products undergo one or more of the following operations followed by wrapping or packaging and. Bivalve molluscs means filter-feeding lamellibranch molluscs Marine biotoxins means poisonous substances accumulated by bivalve molluscs. shelling. and all mammals. mincing or processing. live tunicates and live marine gastropods. slicing. after preparatory work such as bleeding. in particular as a result of feeding on plankton containing toxins. such as gutting. Freezer vessel means any vessel on board in which freezing of fishery products is carried out. slicing. Conditioning means the storage of live bivalve molluscs coming from class A production areas. Fresh fishery products means unprocessed fishery products.LEGISLATION Fishery products means all seawater or freshwater animals (except for live bivalve molluscs. parts and products of such animals. Prepared fishery products means unprocessed fishery products that have undergone an operation affecting their anatomical wholeness. filleting. to remove mud.
4. the mark must be oval shaped and include the abbreviation CE.4. EÜ. the code would be UK. a new mark must be applied to the product. please refer to Regulation (EC) 853/2004. ES. EF. In this case.4. and the characters easily decipherable and be clearly displayed. EK.4.2.1 Identification marking An identification mark needs to be applied following the requirements of Annex II. when a product's packaging and/or wrapping is removed or is further processed in another establishment. (iii) Additionally.2 Form of the identification mark (i) The mark must be legible and indelible. If an establishment manufactures both food to which Regulation 853/2004 applies and food to which it does not. For the UK. the new mark must show the approval number of the establishment where this occurred.FISH AND SEAFOOD 9. EG. EB or WE. (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. Extracts of the requirements of Annex II. EO. 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. Section I of Regulation (EC) 853/2004 are given as follows: 9. 198 . EC. (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. EK.2 Fishery product requirements 9. for other codes. EY. Section I of Regulation (EC) 853/2004. the food business operator may apply the same identification mark to both types of food.1 Application of the identification mark (i) The identification mark must be applied before the product leaves the establishment of production.2.1. the mark must show the approval number of the establishment. (ii) However. (iv) If the identification mark is applied in an establishment within the Community.2.1. 9.
C and D. the wrapping or the packaging. for full requirements. or be printed on a label affixed to the product. (iii) In the case of liquid. Extracts of the requirements of Annex III. It may also be a tag made of a resistant material affixed in such a way that it cannot be removed. the approval number of the establishment and the abbreviation for the community where appropriate.2. wrapping or packaging in another establishment. Chapter IV. (iv) If products of animal origin are placed in a package destined for direct supply to the final consumer. (v) When the mark is applied directly to products of animal origin. and fishery products carried in bulk. echinoderms. a) requirements for establishments.4. the mark may be applied to the outer surface of the container or packaging. Chapter III. Section VIII of Regulation 853/2004. including vessels. (ii) For products of animal origin that are placed in transport containers or large packages. specifically relating to fishery products are given below. the colourings used must be EC authorised colours. The requirements of this section accompany those in Regulation (EC) 852/2004.LEGISLATION 9. reference should be made to the actual Regulation. as amended. the requirements accompany those of Annex II of Regulation (EC) 852/2004 199 . Part A and Chapter V apply to retail. and are intended for further handling. Parts A.3 Method of marking (i) The mark is to be applied directly to the product. 9.2. it is sufficient to apply the mark to the exterior of that package only. granulate and powdered products of animal origin carried in bulk. they would need to be obtained following Section VII on live bivalve molluscs. tunicates and marine gastropods when placed on the market live. With the exception of Chapters I and II. an identification mark is not necessary if accompanying documentation contains the name of the country or its two-letter code.1. its wrapping or the packaging. it applies to those animals when not placed on the market live. Therefore.2 Hygiene requirements for fishery products The regulation details specific hygiene requirements for fishery products.4.e. This section does not apply to bivalve molluscs. i. involved in primary production and such operations accompany the needs of Annex I of Regulation (EC) 852/2004 b) for other establishments including vessels. processing.
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. removing fins.FISH AND SEAFOOD c) for the water supply. refrigeration and wrapping.) 9. within fish farms on land) (ii) the transport of fishery products that have not been substantially altered (including live fishery products. clean seawater may be used for the handling and washing of fishery products. A. Requirements for vessels designed and equipped to preserve fresh fishery products for more than 24 hours 200 .. Structural and equipment requirements refer to the following points 1.1 Chapter I .2.e. bleeding. the production of ice used to chill fishery products and the rapid cooling of crustaceans and molluscs after their cooking. (a) primary production covers the farming. surfaces that contact fishery products need to be made from cleanable corrosion-resistant material. heading. if a vessel has water coming in that is used with fishery products. it must be placed somewhere that will not contaminate the water supply. in that record keeping requirements stated by point 7 in Part A of Annex I of Regulation 852/2004 do not apply to operators involved in small-scale coastal fishing (as defined through Regulation (EC) 1198/2006) if the activity is under 24 hours. this supplements the requirements of Annex II. or to handle or process them after harvesting. There is a derogation from point a. 2. they also include (i) the transport and storage of fishery products that have not been substantially altered (including live fishery products. and (b) related operations include any of the following operations if done on board fishing vessels: slaughter. Requirements for all vessels Vessels should be made in a way that does not cause contamination with substances such as fuel and oil etc. Chapter VII of Regulation 852/2004. i. gutting.4. from the place of production to the first establishment of destination. fishing and collection of live fishery products with a view to their being placed on the market. The definition of fishery products covers primary production as well as its associated operations.2.
and allow the monitoring and. tanks or containers for the storage of fishery products at the temperatures laid down in Chapter VII. (ii) Refrigeration equipment needs to keep fishery products in storage holds at not more than -18 °C. (iii) In vessels equipped for chilling fishery products in cooled clean seawater. Also. Such devices must achieve a chilling rate that ensures that the mix of fish and clean seawater does not exceed 3 °C six hours after loading. 3. 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.LEGISLATION (i) Such vessels need to have holds. (v) Special equipment for disposing of waste/ inedible fishery products into the sea or a watertight tank and allocation of separate areas. where necessary. 4. A temperature recording device must be present and placed where the temperature in the hold is the highest. any waste processing unit needs to have a separate hold to store the waste. (iv) There needs to be somewhere for storing packaging materials and this is to be separate from product preparation and processing 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. Holds and containers used for storing fishery products must allow its preservation under adequate hygiene conditions and. and not more than 0 °C after 16 hours. where necessary. tanks must incorporate devices that allow a uniform temperature throughout the tanks. (iii) The above requirements for holds in vessels designed and equipped to preserve fresh fishery products for more than 24 hours must be met. recording of temperatures. 201 . Requirements for freezer vessels (i) These need to contain freezing equipment in order to achieve a core temperature of no more than -18 °C. to ensure that melt water does not stay in contact with the products. Storage areas for the finished products also need to be sufficiently large and cleanable.
c) Factory vessels that freeze fishery products must have equipment meeting the requirements for freezer vessels as stated above. c) Fishery products. Chapter II . Parts that may present a danger to public health are to be removed quickly and kept away from products for human consumption. B. The brine must not be a source of contamination. Handlers may use spiked instruments to move large fish or fish that might injure them. d) Any gutting or heading of fish on board is to be done hygienically and quickly after capture. landed as soon as possible. provided no damage occurs to the flesh. Livers and roes for human consumption must be preserved under ice (temperature to approach that of melting ice) or be frozen. 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 . must be chilled or.Requirements during and after landing 9. where fishery products are displayed for sale. chilled and wrapped. other than those kept alive. if this is not possible.2.4.2. (ii) In the case of food business operators responsible for auction and wholesale markets etc. b) Factory vessels on board which crustaceans and molluscs are cooked. and products washed thoroughly.2 (i) Unloading and landing equipment that comes into contact with fishery products needs to be made of material that is easily cleanable. b) Fishery products are to be handled and stored in a way that prevents the flesh bruising. the product must be kept at no more than -9 °C. 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.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. 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. e) In cases where whole fish intended for canning is frozen in brine. do not have to meet the above requirements if no other form of handling or processing takes place on board such vessels.
packaged and chilled as quickly as possible after preparation. They should be wrapped.LEGISLATION consumption.Requirements for establishments. C.2. including vessels. (iii) Whole and gutted fresh fishery products can be transported and stored in cooled water on board vessels.2. B. Additionally. (ii) Heading and gutting must be carried out hygienically. Gutting should ideally be done quickly once products have been caught or landed and afterwards be washed thoroughly. points 1 and 2. C. Fillets and slices must then be removed from work tables as soon as possible after preparation. Packaged fresh fishery products must be chilled to a temperature approaching that of melting ice. 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. 9. dispatched. and where necessary.3 Chapter III . Requirements for mechanically separated fishery products (i) Raw materials used must satisfy the following 203 . 854/2004. handling fishery products A. Chapter I. Requirements for fresh fishery products (i) Where chilled or unpackaged products are not distributed. 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. (iv) Filleting and cutting is to be carried out in a way that avoids contamination or spoilage of fillets or slices. they must be stored under ice in appropriate facilities. there are requirements that such premises should be well lit and vehicles emitting fumes should not be able to access the premises. 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. part I.4. (v) Containers for dispatching and storing unpackaged prepared fresh fishery products kept under ice should ensure that melt water does not remain in contact with the products. prepared or processed immediately after arrival at an establishment on land.
raw materials must have no guts. must accompany fishery products referred to above when placed on the market. (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. Requirements concerning parasites (i) The fishery products below must be frozen at no more than .2. (iii) sprat. (ii) The manufacturing process must satisfy the following Mechanical separation is to be carried out immediately after filleting.2.Requirements for certain processed fishery products A.4 Chapter IV .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. Requirements for cooking of crustaceans and molluscs After crustaceans and molluscs are cooked. and whole fish must be gutted and washed beforehand. mechanically separated fishery products must be frozen as quickly as possible or incorporated into a product intended for freezing. cooling must continue until a temperature 204 .4. If no other method of preservation is used. stating the type of process they have undergone. and (c) marinated and/or salted fishery products. 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. Once produced.FISH AND SEAFOOD Only whole fish and bones after filleting may be used to produce mechanically separated fishery products. if the processing is insufficient to destroy nematode larvae. (b) fishery products from the following species. 9. they must be cooled rapidly. (ii) mackerel. except when supplied to the final consumer. D. or a stabilising treatment. (iv) (wild) Atlantic and Pacific salmon. (iii) A document from the manufacturer.
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. 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. (ii) The production process for fish oil must ensure that all raw material intended for the production of crude fish oil is subject to a treatment including (where relevant) heating.2. 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). 9. the following conditions need to be met: A. processing. B.4. raw materials need to be chilled as soon as possible and should meet the temperature requirements in Chapter VII. cooked products are to be frozen or chilled to the temperatures stated in Chapter VII. However.LEGISLATION approaching that of melting ice is reached.5 Chapter V . separation. Histamine Histamine limits must not be exceeded (limits are set in Regulation (EC) No. refining and purification steps before being placed on the market for the final consumer. 2074/2005). 2073/2005.Health standards for fishery products In addition to compliance with microbiological criteria laid down in Regulation (EC) No. 2073/2005). 2074/2005. Shelling/shucking is to be done hygienically. 205 . Requirements for fish oil intended for human consumption (i) In relation to the raw materials used in their preparation and their transport. pressing.2. and after this. centrifugation. and the raw material is processed within 36 hours after loading. C. 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. B.
in particular Ruvettus pretiosus and Lepidocybium flavobrunneum.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. In the case of whole frozen fish in brine intended for the manufacture of canned food.4. (ii) Frozen blocks prepared onboard vessels must be adequately wrapped before landing.FISH AND SEAFOOD D.6 Chapter VI . Parasites Fishery products are to be visually examined so that visible parasites are identified before being placed on the market. 9.2. 9. The requirements for histamine and parasites do not apply to whole fishery products being used directly for preparing fish oil intended for human consumption. and cooked and chilled products from crustaceans and molluscs are to be kept at a temperature close to that of melting ice. prepared and processed fishery products belonging to the family Gempylidae. Fresh. and when fishery products are wrapped on board fishing vessels. Diodontidae and Canthigasteridae. E. Fishery products contaminated with parasites may not be placed on the market for human consumption.4.2. Toxins harmful to human health Fishery products derived from poisonous fish of the following families must not be placed on the market: Tetraodontidae. it should be kept at no more than -9 °C.2. 206 . Molidae. 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. (ii) Frozen fishery products must be kept at a temperature of not more than -18 °C in all parts of the product. The scientific name must accompany the common name on the label. 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.Storage of fishery products (i) Fresh fishery products.7 Chapter VII . thawed unprocessed fishery products.2.
9. The following in relation to TVB-N limits is extracted from the regulation: Unprocessed fishery products shall be regarded as unfit for human consumption where organoleptic assessment has raised doubts as to their freshness and chemical checks reveal that the following TVB-N limits are exceeded: (a) 25 mg of nitrogen/100 g of flesh for the species: Sebastes spp. (ii) If fishery products are kept under ice. (d) 60 mg of nitrogen/100 g of whole fishery products used directly for the preparation of fish oil for human consumption as referred to in the second sub-paragraph of Part B(1) of Chapter IV of Section VIII of Annex III to 207 . Sebastichthys capensis. species belonging to the Merlucciidae family. (c) 35 mg of nitrogen/100 g of flesh for the species: Salmo salar.2. provided that the journey is short and the competent authority allows this.4. The temperature of frozen fishery products must not fluctuate above -15°C. The required temperature for whole frozen fish in brine intended for the manufacture of canned food during transport is not stated. 9. This covers Total volatile basic nitrogen (TVBN) limits and how to determine them. Note: (i) Frozen fishery products do not need to be kept at a temperature of -18 °C if they are being transported from a cold store to an approved establishment to be thawed on arrival for the purposes of preparation and/or processing. species belonging to the Gadidae 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.8 Chapter VIII .LEGISLATION (iii) Fishery products kept alive are to be kept at a temperature which will not adversely affect food safety or its viability.2. (b) 30 mg of nitrogen/100 g of flesh for the species belonging to the Pleuronectidae family (with the exception of halibut: Hippoglossus spp.Transport of fishery products The above points for Chapter VII apply in relation to transport.2. melt water must not remain in contact with the products.). 2074/2005.4. Helicolenus dactylopterus.
specifically relating to bivalve molluscs are given below.4. they supplement the requirements of Annex I to that Regulation. this Section also applies to live echinoderms.FISH AND SEAFOOD Regulation (EC) No 853/2004.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. The reference method to be used for checking the TVB-N limits involves distilling an extract deproteinised by perchloric acid as set out in Chapter III For distillation apparatus and routine methods which may be used to check the TVB-N limits. however. where the raw material complies with points (a). Apart from the provisions on purification. The requirements of this Section accompany those laid down in Regulation (EC) No 852/2004: (a) In the case of operations that take place before live bivalve molluscs arrive at a dispatch or purification centre.3. The information required in the registration document is given in point 4 and includes the health status of the production area. (b) In the case of other operations. Member States may set limits at a higher level for certain species pending the establishment of specific Community legislation. reference should be made to the actual regulation. Section VII of Regulation 853/2004. taken from at least three different points and mixed together by grinding. as amended. 9. The sample must consist of about 100 g of flesh. tunicates and marine gastropods. 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. shellfish species etc. Extracts of the requirements of Annex III. Chapters V. VI.1 Chapter I . VIII and IX. Live bivalve mollusc requirements - 9. they supplement the requirements of Annex II to that Regulation. (b) and (c) of Part B(1) of that Chapter. The copy of 208 .4. Chapter IX applies to pectinidae harvested outside those areas. for full requirements.3 The regulation details specific hygiene requirements for bivalve molluscs. please refer directly to the regulation. 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.
Other requirements such as conditions for relaying in order to allow optimal conditions for purification and for permanent record keeping are also specified. Requirements for production areas (i) The competent authority classes (classed A.Class A means they can be placed directly on the market .LEGISLATION document must be kept for at least 12 months post dispatch or receipt or longer as stated by the competent authority. C.4. 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. 9. (ii) Provided the requirements of chapter V are met. 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.3.Class C means relaying over a long time before it can be placed on the market For bivalve molluscs not subject to Class B or C treatments. Transport needs to allow sufficient drainage for best survival conditions. B. 209 . B or C) production areas with fixed locations and boundaries for where live bivalve molluscs may be harvested and this must be complied with.Class B means treatment in a purification centre or after relaying is done before it can be placed on the market . poles or other fixed means are to identify the boundaries of the sites. collection from: .2 Chapter II . 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. processing or relaying.Hygiene requirements for the production and harvesting of live bivalve molluscs A.
Requirements for purification centres There are requirements for live bivalve molluscs to be washed. (iv) yessotoxins. Before dispatch.4. the levels of marine biotoxins should not exceed the following limits. 9. 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. for: (i) paralytic shellfish poison (PSP). containers and a label to identify purification. 160 micrograms of okadaic acid equivalents per kilogram.4. 800 micrograms per kilogram. Requirements for dispatch centres Handling should not result in contamination to any degree.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 if there are many batches of live bivalve molluscs.3.FISH AND SEAFOOD 9. (ii) amnesic shellfish poison (ASP). the purification system to allow them to resume /maintain filter-feeding activity.Hygiene requirements for purification and dispatch centres A.3. 210 .3. 1 milligram of yessotoxin equivalent per kilogram. and there are requirements for tanks and water storage containers.Structural requirements for dispatch and purification centres Premises on land should not be subject to flooding by tides etc. B.5 Chapter V .3 Chapter III .4. dinophysistoxins and pectenotoxins together.4 Chapter IV .
Registration document requirements also apply. 9. dispatch centre or processing establishment.4.3.Identification marking and labelling In addition to general identification marking (contained in 9.3.8 Chapter VIII . (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. (ii) Once the live bivalve molluscs have been packed for retail sale and have left the dispatch centre.Wrapping and packaging of live bivalve molluscs (i) Oysters must be wrapped or packaged with the concave shell downwards.4.6 Chapter VI . the label. Also.9 Chapter IX . 160 micrograms of azaspiracid equivalents per kilogram. 211 .4. 9.4. the date of minimum durability may be replaced by the words ‘these animals must be alive when sold’. (Note: testing methods for detecting marine biotoxins are detailed in Annex III of Regulation (EC) 2074/2005) 9. the species of bivalve mollusc and date of packaging (at least the date and month) are required. Finally.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. 9.1).3. The requirements for production areas as stated earlier apply to pectinidae and these can only be placed on the market through a fish auction. must be waterproof.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.2. including the identification mark.7 Chapter VII . by way of derogation from directive 2000/13/EC.4. Requirements of Chapters III and IV are to be complied with.3. they cannot be re-immersed in or sprayed with water.LEGISLATION (v) azaspiracids.
The official controls include audits of good hygiene practices and hazard analysis and critical control point (HACCP)-based procedures. they derive from: (i) poisonous fish. or - - 212 . physical or microbiological checks or checks for parasites have shown that they are not in compliance with the relevant Community legislation. Chapter V of Regulation 853/2004 apply. In this regulation. (ii) fishery products not complying with the requirement concerning biotoxins. they contain in their edible parts contaminants or residues in excess of the limits laid down in Community legislation or at levels where the calculated dietary intake would exceed the acceptable daily or weekly intake for humans. tunicates or marine gastropods containing marine biotoxins in total quantities exceeding the limits referred to in Regulation (EC) No 853/2004. Therefore the health standards for fishery products as previously stated in Annex III. chemical.5 Regulation (EC) No. Fishery products in particular must comply with the requirements of Annex III of the regulation. or (iii) bivalve molluscs.1 Regulation (EC) 854/2004 Annex III Chapter III states that fishery products are to be declared unfit for human consumption if: organoleptic. general principles for official controls in respect of all products of animal origin falling within the scope of the regulation are given. echinoderms. 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.5. 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. Section VIII.FISH AND SEAFOOD 9. This refers to official controls or production and placing on the market of fishery products. 9. The requirements refer to hygienic conditions and regular inspections both on land and at port to ensure compliance of temperature and other requirements.
Official controls concerning live bivalve molluscs from classified production areas A. Monitoring of classified relaying and production areas This is to ensure there is no malpractice. A summary is given as follows: 9. 4.5. - - - - B. Live bivalve molluscs from Class C areas must not exceed 46.5. 000 E. Regulation (EC) 854/2004. The reference method for this analysis is the five-tube.2 This states official controls for the production and placing on the market of live bivalve molluscs. Live bivalve molluscs from Class B areas must not exceed. Live bivalve molluscs from class A areas may be collected for direct human consumption provided health standards are met. chapter V is reinforced. chapter II on hygiene requirements for the production and harvesting of live bivalve molluscs in Regulation (EC) 853/2004. that microbiological criteria and chemical contaminants amongst other requirements are met. coli per 100 g of flesh and intravalvular liquid. coli per 100 g of flesh and intravalvular liquid. 213 . three dilutions Most Probable Number (MPN) test specified in ISO 16649-3. live tunicates and live marine gastropods. in 90% of the samples. three dilutions MPN test specified in ISO 16649-3. section VII. In the remaining 10 % of samples. as previously stated.1 Chapter II . live bivalve molluscs must not exceed 46.LEGISLATION - the competent authority considers that they may constitute a risk to public or animal health or are for any other reason not suitable for human consumption. The reference method for this analysis is the five-tube. Requirements for the competent authority are stated in relation to the classification or a production or relaying area is stated.2. coli per 100 g of flesh and intravalvular liquid. Classification of production and relaying areas: This section reinforces that specified in Annex III. Also. live echinoderms.600 E. Sampling plans and frequency are required.000 E. Annex II 9.
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. E. 2073/2005 on Microbiological Criteria for Foodstuffs Regulation (EC) No.6 Regulation (EC) No. 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. 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.Official controls concerning pectinidae harvested outside classified production areas These are to be carried out in fish auctions.FISH AND SEAFOOD C. 2073/2005. 9.3 Imports By way of derogation from Article 11(1) of Regulation (EC) No 854/2004.2 Chapter III . F. boundaries or class of a production area or its closure. Decisions after monitoring There are specific requirements whereby if sampling results indicate that health standards for molluscs are exceeded or there is a risk to human health. dispatch centres and processing establishments in order to confirm that health standards and other specific requirements are met.2. 214 . 9. establishes microbiological criteria for a range of foods.5. Food business operators' own checks Controls that food business operators have done are considered by the competent authority when classifying. Procedures for re-opening a closed production area are stated D. the competent authority is to close the production area.5. opening or closing a production area. 9. which has applied since 1 January 2006.
ensuring that foods being placed on the market do not pose a risk to human health. 9.6. based on the absence. a batch of foodstuffs or a process. Two kinds of criteria have been established: food safety criteria. with chapter 2.2 Process hygiene criteria Chapter 2 focuses on process hygiene criteria. per unit(s) of mass. volume.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.I.6. 215 . and the legislation applies to all businesses involved in food production and handling. 9. The criteria are outlined in Table 9. The relevant criteria are outlined in Table 9. area or batch.4 referring to fishery products. The definition of ‘microbiological criterion’ means a criterion defining the acceptability of a product. presence or number of microorganisms.LEGISLATION The aim of this legislation is to complement food hygiene requirements. applying to products placed on the market and process hygiene criteria that are applied during the manufacturing process.II. and/or on the quantity of their toxins or metabolites.
other than those intended for infants and for special medical purposes(4) (8) L.3 Ready-to-eat foods unable to support the growth of L. monocytogenes. monocytogenes 5 0 Absence in 25 g (7) FISH AND SEAFOOD 216 5 0 100 cfu/g 5 0 Absence in 25 g 1.I Food safety criteria Sampling plan(1) n 10 5 Absence in 25 g EN/ISO 11290-1 c m M Products placed on the market during their shelf life Limit(2) Analytical reference method(3) Stage where the criterion applies Food category Microorganisms 1. 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. who has produced it EN/ISO 11290-2 (6) Products placed on the market during their shelf life 1. monocytogenes 1.2 Ready-to-eat foods able to support the growth of L. monocytogenes.1 Ready-to eat foods intended for infants and ready-to-eat foods for special medical purposes (4) 5 0 100 cfu/g (5) EN/ISO 11290-2 (6) L.TABLE 9.16 Cooked crustaceans and molluscan shellfish Salmonella EN/ISO 6579 Products placed on the market during their shelf life . monocytogenes L.
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.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 1.25 Live bivalve molluscs and live echinoderms. tunicates and gastropods 1(16) 0 230 MPN/100 g of flesh and intravalvular liquid 100 mg/kg 200 mg/kg HPLC (19) ISO TS 16649-3 Salmonella 1. tunicates and gastropods 9 (18) 2 E. manufactured from fish species associated with a high amount of histidine (17) Histamine .17 Live bivalve molluscs and live echinoderms.27 Fishery products which have undergone enzyme maturation treatment in brine.
Single samples may be taken at retail level. products with a shelf-life of less than five days shall be automatically considered to belong to this category. Scombresosidae. This criterion shall apply to products before they have left the immediate control of the producing food business operator. 5 This criterion shall apply if the manufacturer is able to demonstrate. Relevance of matrix effect in determination of biogenic amines in 6 7 8 15 16 17 18 19 218 .92. wine. uncut and unprocessed vegetables and fruits. that the product will not exceed the limit 100 cfu/g throughout the shelf life. AOAC Internat.. biscuits and similar products. — fresh. The most recent edition of the standard shall be used.. honey and confectionery. Bouquelet S. c = number of sample units giving values between m and M For point 1.1 . 2. Engraulidae. 1 ml of inoculum is plated on a Petri dish of 140 mm diameter or on three Petri dishes of 90 mm diameter. — bread. live bivalve molluscs. Bouquelet S. that the product will not exceed the limit of 100 cfu/g throughout the shelf life. Valle M. Duflos G. coli is used here as an indicator of faecal contamination. sugar. 79.25. Particularly fish species of the families: Scombridae. to the satisfaction of the competent authority. J. m = M.1. 1996. Dervin C.. when recontamination is not possible after this treatment (for example. 43-9. spirits and similar products. cider. Products with pH ≤ 4. 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. Coryfenidae. Malle P.4 or aw ≤ 0. 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. In such a case the presumption laid down in Article 14(6) of Regulation (EC) No 178/2002. according to which the whole batch is to be deemed unsafe. monocytogenes. — — bottled or packed waters. Malle P. soft drinks. shall not apply. Other categories of products can also belong to this category. when he is not able to demonstrate. to the satisfaction of the competent authority. E. beer. including cocoa and chocolate products..94. excluding sprouted seeds. Clupeidae.0 and aw ≤ 0.. Assay of biogenic amines involved in fish decomposition. References: 1. subject to scientific justification.FISH AND SEAFOOD 1 2 3 4 n = number of units comprising the sample. products heat treated in their final package). Pomatomidae. products with pH ≤ 5. A pooled sample comprising a minimum of 10 individual animals.
if all the values observed indicate the absence of the bacterium. tunicates and gastropods in relation to testing E. Interpretation of the test results relating to Table 9.I The limits given refer to each sample unit tested. if all the values observed are ≤ the limit. if all the values observed indicate the absence of the bacterium. J. Histamine in fishery products from fish species associated with a high amount of histidine: — satisfactory. excluding live bivalve molluscs and live echinoderms. no values observed exceed the limit of M. L. Salmonella in different food categories: — satisfactory. L. — unsatisfactory. monocytogenes in ready-to-eat foods able to support the growth of L. (1) 219 . a maximum of c/n values observed are between m and M 3. — unsatisfactory. coli. — unsatisfactory. monocytogenes in other ready-to-eat foods: — satisfactory. L. monocytogenes before the food has left the immediate control of the producing food business operator when he is not able to demonstrate that the product will not exceed the limit of 100 cfu/g throughout the shelf life: — satisfactory. if all the values observed indicate the absence of the bacterium. if any of the values are > the limit. The test results demonstrate the microbiological quality of the batch tested(1). the mean value observed is ≤ m 2. 82. if the presence of the bacterium is detected in any of the sample units. 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.LEGISLATION plaice (Pleuronectes platessa) and whiting (Merlangus merlangus. 1999. where the limit refers to a pooled sample. if the following requirements are fulfilled: 1. if the presence of the bacterium is detected in any of the sample units. — unsatisfactory. — unsatisfactory. AOAC Internat. 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. monocytogenes in ready-to-eat foods intended for infants and for special medical purposes: — satisfactory. if the presence of the bacterium is detected in any of the sample units. 1097-101.
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 2.4.1 Shelled and E. 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 .TABLE 9.
— acceptable. 9. if a maximum of c/n values are between m and M. had not been 221 . coli in shelled and shucked products of cooked crustaceans and molluscan shellfish: — satisfactory. the defendant will need to prove that the food was on service or display. which overrides the 8 °C requirement.7 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.LEGISLATION 1 n = number of units comprising the sample. 14 details temperature control requirements for foods in general.K) Temperature control requirements 9.1 In the U.I. — acceptable. and particularly that perishable foodstuffs must not be kept at above the maximum recommended storage temperature. 2 Interpretation of the test results relating to Table 9. 2006 No. if all the values observed are ≤ m.7. The test results demonstrate the microbiological quality of the process tested. S. if all the values observed are ≤ 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. if one or more of the values observed are > M or more than c/n values are between m and M. For example. with chill holding tolerance periods. Coagulase-positive staphylococci in shelled and cooked crustaceans and molluscan shellfish: — satisfactory. — unsatisfactory. tolerance periods for chill-holding of foods and hot-holding variations. The regulations prescribe a chilled food holding temperature of 8 °C or less.K. S.II The limits given refer to each sample unit tested.I. c = number of sample units giving values between m and M The most recent edition of the standard shall be used. 2006 No. E. and the rest of the values observed are ≤ m. and the rest of the values observed are ≤ m. Schedule 4 of the Food Hygiene (England) Regulations 2006. 14 (Hygiene requirements specific to the U. The regulations provide for defences in relation to upward variations of the 8 °C temperature. The defendant may be required to produce well-founded scientific proof to support his claims. 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. — unsatisfactory.
or at a temperature above 63 °C and to reheat food to a temperature of at least 82 °C (The Food Hygiene (Scotland) Regulations 2006. the Food Standards Agency has published guidance notes on the requirements of the EU hygiene and microbiological criteria regulations which can be accessed at the following link: http://www. or. the period for which it can be expected to remain fit for sale when kept in a manner consistent with food safety. 3). It should be noted that the temperature control requirements as detailed in Schedule 4 of the Food Hygiene (England) Regulations 2006 (S.S. For other food. 9.K. such as that below. S. to or from premises (including vehicles) at which the food was to be kept at or below 8 °C or the recommended temperature. 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. preparing or processing the food recommending that it be kept at or below a specified temperature between 8 ºC and ambient temperatures. Recommended temperature: a specified temperature that has been recommended in accordance with a food business responsible for manufacturing.uk/foodindustry/guidancenotes/hygguid/fhlguidance/ 222 . 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.gov.8 Guidance In the U.I. including: Shelf life: where the minimum durability or 'use by' indication is required according to Regulation 20 or 21 of the Food Labelling Regulations 1996 (form of indication of minimum durability and form of indication of ‘use-by date’) the period up to and including that date.I. 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. The permitted reasons are given below: to facilitate handling during and after processing or preparation the defrosting of equipment.food. was kept at above 8 °C or the recommended temperature for an unavoidable reason.FISH AND SEAFOOD previously put on display at more than 8 °C and had been kept there for less than four hours. 2006 No. and was kept at above 8 °C or the recommended temperature. 2006 No. Alternatively. Schedule 4 of the Food Hygiene (England) Regulations 2006 contains several definitions.
14.2004. 9.12. Regulation (EC) No. 223 .2006. L226. L226. 1663/2006. The revised text of Regulation (EC) No.9 Other Relevant Legislation The following regulations about fish designations need to be considered: • Regulation (EC) No 104/2000 on the common organisation of the markets in fishery and aquaculture products Regulation (EC) No 2065/2001 laying down detailed rules for the application of Council Regulation (EC) No 104/2000 as regards informing consumers about fishery and aquaculture products • Note: Requirements for preserved fish are laid down elsewhere.6.2004.6. namely. 2073/2005 on microbiological criteria for foodstuffs (OJ No.2005. as read with the corrigenda at OJ No.10 Further Reading Regulation (EC) No. 1. 25. 83) as amended by Regulation Nos 882/2004. 30. 852/2004 of the European Parliament and of the Council on the hygiene of foodstuffs (OJ No.2004. L139. Regulation (EC) No. 2074/2005. Regulation (EEC) 2136/89. 2076/2005. 25. 853/2004 is now set out in a Corrigendum (OJ No. 3) as amended by Regulation (EC) No. 2076/2005. 2075/2005 and Regulation (EC) No. 55). and Regulation (EEC) 1536/92 laying down common marketing standards for preserved tuna and bonito.6.4.LEGISLATION 9. Regulation (EC) No. 25. L338. 206). L283.2004. as amended. 2076/2005. 1019/2008 and as read with Regulation 2073/2005. these are not be discussed here. 1791/2006 and 1020/2008 and as read with EC Directive 2004/41. 22) as amended by Regulation (EC) Nos. laying down common marketing standards for preserved sardines. 853/2004 of the European Parliament and of the Council laying down specific hygiene rules for food of animal origin (OJ No. 854/2004 is now set out in a Corrigendum (OJ No. L226. 1662/2006. 1688/2005. 1).2004. 852/2004 is now set out in a Corrigendum (OJ No. 62) as amended by Regulation (EC) No. The revised text of Regulation (EC) No. L139. Regulation (EC) No. Commission Regulation (EC) No.4. 2076/2005.10. The revised text of Regulation (EC) No. 2074/2005 and Regulation (EC) No. L139. 2074/2005. 30. 854/2004 of the European Parliament and of the Council laying down specific rules for the organisation of official controls on products of animal origin intended for human consumption (OJ No. Regulation (EC) No. 30. Regulation(EC) No. 22. 2074/2005.2004. 1791/2006 and 1021/2008 and as read with EC Directive 2004/41.4. 1441/2007.
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.2001.10. (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 338.1. 22. 6) 224 . 22. 27 Commission Regulation (EC) No 2076/2005 laying down transitional arrangements for the implementation of Regulations (EC) No 853/2004.12. 21. 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 854/2004.2005.12. 83) Regulation (EC) No 104/2000 on the common organisation of the markets in fishery and aquaculture products (OJ L 17. as amended by Commission Regulation (EC) No 1022/2008 OJ L 338. 22) Commission regulation (EC) No 2065/2001 laying down detailed rules for the application of Council Regulation (EC) No 104/2000 as regards informing consumers about fishery and aquaculture products as amended by Commission Regulation (EC) No 1792/2006 of 23 October 2006 (OJ L 278. 23.2000.2005.
under optimal incubation temperatures.3 minutes.1.1 10.10.1.0 .4 Heat resistance Aeromonads are sensitive to moderately high temperatures.3.10 (3).1. The optimum growth temperature is 28 °C (2). Aeromonas has the potential to grow over a pH range of ca 4 .0 x 184.108.40.206 Aeromonas spp. Morphology Gram-negative rods.000-fold increase in number is observed after one week in food systems (1).45 °C (1).1. 225 . and D-values at 45 and 51 °C have been reported as 29.1.5 pH Under otherwise optimal conditions. 10. At 5 °C. At pH 4. 10.5 minutes.3 . PATHOGEN PROFILES 10.1.2 Oxygen requirements Facultative anaerobe. The minimum pH value for growth is thought to be pH 5. 0.3 Temperature Aeromonas grows at temperatures of 2 . a 10 to 1. and 2.5 µm 10. 10. no growth is observed (1). Some death will occur at this pH if the temperature is 4 °C. respectively (1).
at 85 ºC for 5 minutes.22. There are four main groupings of the organism. Group I will not grow at temperatures of 10 ºC or less.7 . but the spores of this organism are more so. E and.2. spore-forming rods. types B and F can be either.2 Clostridium botulinum Seven different types of C. which may be inactivated by heating at 80 ºC for 20 .30 minutes. botulinum are not particularly heat resistant.2. or at 90 ºC for a few seconds.2. to a lesser extent. botulinum is an obligate anaerobe. and is sensitive to > 4. and groups I and II are those responsible for the botulism cases. Types A.FISH AND SEAFOOD 10. 10.2 Oxygen requirements Although C. botulinum types produce heat labile toxins. 10. being capable of slow growth and toxin production at low temperatures .2. but Group II strains are psychrotrophic. Thus an aerobically packed product may not support the growth of the organism on the surface.even as low as 3 ºC (7). 226 .3 Temperature All strains of C.1.6 Aw/Sodium chloride Growth of Aeromonas is optimal in the presence of 1 .4 Heat resistance The vegetative cells of C. All C.0 µm. A to G. botulinum grow reasonably well in the temperature range of 20 45 ºC. 0. B.1 Morphology Gram-positive. botulinum is prevented. It is also important to note that the inclusion of oxygen as a packaging gas cannot ensure that growth of C.5% NaCl (4). but the minimum temperatures required to inhibit Groups I (proteolytic group) and II (non-proteolytic group) are different. 10.4 x 1. many foods that are not obviously ‘anaerobic’ can provide adequate conditions for growth. botulinum are known.2% sodium chloride (NaCl) (1). 10.2.5 . All type A strains are proteolytic and type E strains are usually non-proteolytic. 6). 10. but the interior of the food may do so. F are the types that are responsible for most cases of human botulism (5. forming at least seven different toxins.
botulinum depends on solute. botulinum spores The most heat resistant spores of Group I C.21 minutes) (9).2 Oxygen requirements C.2. vacuum packed. Note also that if humectants other than salt are used.6 Aw/ Sodium chloride The minimum water activity (aw) for growth of C. 0. but under optimum growth conditions 10% (w/w) NaCl is required to prevent growth of Group I.6 and pH 5. where oxygen levels are reduced. 10. botulinum are produced by type A and proteolytic B strains (D121ºC=ca 0. pH and temperature. These values have been established under carefully controlled laboratory conditions.9 x 2.10.7 Characteristics of C.2. 10.85 ºC) and their ability to grow at refrigeration temperatures necessitates their control in foods capable of supporting their growth (e.g. respectively (8. 10. perfringens . In commercial situations. parcooked meals with pH value > 5.1.5 pH The minimum pH for the growth of proteolytic and non-proteolytic strains is pH 4. they may survive mild heat treatments (70 . 227 . It will not. However. and 5% (w/w) NaCl is necessary to prevent growth of Group II organisms.3 10. grow on the surface of foods unless they are vacuum or gas-packed.PATHOGEN PROFILES 10. 9). 11)).94 for Group I and 0.is an obligate anaerobe. safety margins must be introduced to allow for process variability.2. the minimum aw for growth may be lower.like other clostridia . particularly by cooking.3. 10.0.0 µm.3 . The organism will grow well in the centre of meat or poultry dishes.1 Clostridium perfringens Morphology Gram-positive spore-forming rods.3.0 and aw > 0.0 . therefore.97 for Group II (5). These concentrations correspond to limiting aw of 0. The spores of Group II (non-proteolytic/psychotrophic) strains are less heat resistant than Group I strains.97) (10.
At cold temperatures. 16).4 Heat resistance Exposure to a temperature of 60 ºC or more will result in the death of vegetative cells of C.50 ºC. perfringens has the potential ability to grow within the temperature range 15 . vegetative cells die rapidly (12). perfringens spores The spores of C. pH. Salt concentrations of 6 .3. Recorded heat 228 . 10. although C. will survive greater extremes of pH (and aw) (12. perfringens is not a tolerant organism with respect to pH. perfringens strains. death of the vegetative cells of this organism usually occurs rapidly above this temperature (12.3.5 minutes (17). The lowest aw recorded to support the growth of C. As in the case of other factors limiting the growth/survival of this organism. Some studies indicate that the presence of 3% NaCl delays growth of C.FISH AND SEAFOOD 10. perfringens in vacuum packed beef (18). 10.45 ºC.3 Temperature The most significant characteristic of C. perfringens in roast beef at 60 ºC is 14. perfringens can vary quite considerably in their heat resistance.it is destroyed by heating at 60 ºC for 10 minutes (14.7 Characteristics of C.8% inhibit growth of most C.3. perfringens appears to be 0.97 (glycerol and sucrose respectively) depending on the solute used to control aw (13. While some growth can occur at 50 ºC. 10. 10. type of solute. (It is unusual for spores to be formed in foods after the growth of this organism (14)). It grows best at pH values between 6 and 7 (the same pH as most meats). 15.10 ºC.3.5 pH C. In addition.6 Aw/Sodium chloride C. 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. however. 18). perfringens. perfringens in relation to food safety is the organism’s ability to grow extremely rapidly at high temperatures. lower concentrations may be effective in combination with other factors. perfringens is not tolerant of low water activities. the limits for aw are affected by temperature. Under otherwise ideal conditions. 0 . the enterotoxin is not heat resistant . Spores. very limited growth may occur at pH values over the range pH ≤ 5 and ≥ 8.3. Its optimum temperature for growth is 43 . The D-value for C.3. etc.93 and 0. 13). depending on strain and other conditions. 13).
albeit slowly .especially at refrigeration temperatures. 229 . It has been reported to have slightly greater heat resistance than certain other foodborne pathogens. however. monocytogenes in crawfish tail meat at 55.98 and 0. 0.64. and temperature.5 x 0.0.19 minutes. 10. and where acetic acid is used as an acidulant.4. it is not a sporeformer. the lowest limit for growth is likely to be considerably higher . 10.2.1 Listeria spp. 10.4 ºC is probably a more likely minimum for growth in foods (19). 60 and 65 ºC were reported as 10. respectively (21).4.3 .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.at refrigeration temperatures down to. Its optimum growth temperature.PATHOGEN PROFILES resistance values at 95 ºC (D-values) range from 17. or even below 0 ºC. growth at low temperatures can be very slow.6 . being potentially capable of growing . however.8 minutes for heat sensitive spores (12).2 has been suggested as the lowest working limit (22).5 . the organism is able to grow at pH values well below pH 5 (pH 4. 10. 10.23.0 µm.4 10. so can be destroyed by pasteurisation. requiring days or weeks to reach maximum numbers.2 Oxygen requirements Aerobe or microaerophilic. monocytogenes is not a particularly heat resistant organism.2. pH < 5. -0. 1.3 is the lowest value where growth has been recorded. The upper temperature limit for the growth of L.4. monocytogenes is reported to be 45 ºC (20).4 . using hydrochloric acid as an acidulant). Under ideal conditions.3 Temperature Listeria monocytogenes is unusual amongst foodborne pathogens in that it is psychrotrophic.4.4 Heat resistance L. However. D-values for L.0 minutes for heat resistant spores to 1. is between 30 and 37 ºC. In foods.4. Morphology Gram-positive short rods.
12% NaCl or more).2 Oxygen requirements Facultative anaerobe.90.5 10. 0. 10. 10.5.5.5.6 Aw/Sodium chloride Plesiomonas can grow well at NaCl concentrations up to 4% (24. 0. It is likely to survive. 10.5 . peritrichous flagella. or even grow.3 Temperature Plesiomonas can grow in the temperature range of 8 .4. 230 .83 (20). The bacterium may survive for long periods at aw values as low as 0.5 pH Plesiomonas has a pH range for growth of pH 4. Morphology Gram-negative short rods. 25).FISH AND SEAFOOD 10. at salt levels found in foods (10 .1 Salmonella spp. monocytogenes is quite tolerant of high NaCl/low water activities.7 x 1.6 Aw/Sodium chloride L.3.1.1 Plesiomonas Morphology Gram-negative.5.97.0 . 25).6.5 (24.8. 10. the optimum being 30 ºC (23).5.5. 10.0 x 1 . It grows best at aw of ≥ 0. 10.0. 10.5 .45 ºC.0 µm.4 Heat resistance Pasteurisation at 60 ºC for 30 minutes has been reported as being effective in killing Plesiomonas shigelloides (24).6 10.3 µm. but has been shown to be able to grow at an aw level of 0.8 .
0.5. Some death will occur at pH values of less than about 4. with a z-value of 4 . Furthermore. temperature and pH. and with an appropriate acid. 10. high fat or low moisture levels (low aw) will reduce the effectiveness of heat treatments. therefore. Salmonellae are quite resistant to drying. D60 ºC values normally range from about 1 to 10 minutes. It is not.6.6.5 .PATHOGEN PROFILES 10.6 Aw/Sodium chloride Where all other conditions are favourable. Growth is slow at temperatures below about 10 ºC.8 . a heat resistant organism. 231 . and appropriate heat treatments must be determined experimentally for low-aw foods.4 Heat resistance Salmonella is not a spore-forming organism. 10.3 Temperature Salmonellae can grow in the temperature range of 7 . some strains are able to grow at temperatures as low as 4 °C (26).5. The optimal pH for Salmonella growth is between 6. The growth of Salmonella is generally inhibited by the presence of 3 .945. Salmonellae are quite resistant to freezing. under otherwise ideal conditions.9. strains vary in their ability to withstand heating. the optimum being 35 .20 times more heat resistant than the average strain of Salmonella at high aw (27). However.5 pH Salmonella has a pH range for growth of pH 3. pasteurisation and equivalent treatments will destroy the organism under normal circumstances. 10.7. Salmonella senftenberg 775W is about 10 .6.6. substrate. However. Salmonella has the potential to grow at aw levels as low as 0.48 ºC.5 ºC. or possibly 0.37 ºC.93 (as reported in dried meat and dehydrated soup). 10.4% NaCl. depending on serotype. although salt tolerance increases with increasing temperature (28). depending on the type of acid and temperature.2 Oxygen requirements Facultative anaerobe.6.
0. fat content. NB: The growth of Staph.7.7. aureus is more limited under anaerobic than aerobic conditions.37 ºC (29). Most strains of staphylococci can grow within the pH range 4. with an optimum of 35 . The following relate to limits for growth only.0 .1 Staphylococcus aureus Morphology Gram-positive cocci.2 Oxygen requirements Facultative anaerobe. but may cause some cell damage (30). 232 .FISH AND SEAFOOD 10.7. 10. In meat macerate. Under most circumstances.7. It can survive well at low temperatures. however.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. aureus than citric acid).3 (optimum 7. 10. 10. 10.4 Heat resistance Heat resistance depends very much on the food type in which the organism is being heated (conditions relating to pH.). Freezing and thawing have little effect on Staph.7. stressed cells can also be less tolerant of heating.2 to 9. 31). aw.9 µm diameter. aw and temperature (sensitivity to acid increases with temperature). etc.20 minutes. the D60 ºC value is 2 .7.5). The limits for toxin production are also narrower than for growth. 0. under otherwise ideal conditions (29. the organism is heat sensitive and will be destroyed by pasteurisation.3 Temperature Under otherwise ideal conditions. As is the case with other bacteria. aureus can grow within the temperature range 7 – 48. Staph. depending on aw.5 ºC.7 10.7 . aureus viability.
8. aureus is unusual amongst food-poisoning organisms in its ability to tolerate low water activities. Staph.6 µm 10.83 .0.7 Limits permitting toxin production 10 . and solute.0 . It can grow over the aw range 0.2.8.6 Aw/Sodium chloride Staph.PATHOGEN PROFILES 10.46 ºC (optimum between 35 and 40 ºC) (very little toxin is produced at the upper and lower extremes) (31) 5. 0. 10.2 Oxygen requirements Facultative anaerobe. 10.1 Vibrio cholerae Morphology Gram-negative curved rods. 10. strain. In general. However. especially vacuum packed foods (33) Temperature: pH: *Aw: Atmosphere: Heat Resistance: Enterotoxins are quite heat resistant.7.5) (29.87 and > 0.86 is the generally recognised minimum in foods (32). An effect of increasing salt concentration is to raise the minimum pH of growth.8.99 little or no toxin production in anaerobically packed foods.7. 34). 233 .>0.10% NaCl.0 (optimum 7.3 Temperature The optimum temperature for growth has been reported as 30 ºC. Staphylococci are more resistant to salt present in foods than other organisms. an aw of 0. aureus can grow in 7 .8 x 1. with a temperature range for growth of 10 . * dependent on temperature. 31) between 0. but certain strains can grow in 20% NaCl.4 .9. pH. In general. heating at 100 °C for at least 30 minutes may be required to destroy unpurified toxin (31.2 .7.43 ºC (35).99 aerobically under otherwise optimal conditions.5 . atmosphere.8 10.
The D-value for V. 10. and it is sensitive to refrigeration temperatures (0 .998. V. cholerae is sensitive to heat at temperatures of >45 ºC. it has a pH range for growth of pH 5 . 38).8.4 Heat resistance V. 10.9.6 (35. with an optimum of 37 ºC (39).5 ºC). under otherwise favourable conditions.0% NaCl (35).1 . cholerae may persist on frozen fish for >180 days (36). 10.2 Oxygen requirements Facultative anaerobe. but only moderately sensitive to freezing (40). cholerae in crab meat homogenate at 60 ºC is 2.1 Vibrio parahaemolyticus Morphology Gram-negative curved rods. respectively. are 5 and 43 ºC. cholerae can grow over the aw range 0.0.65 minutes (35). there is evidence that V.3 Temperature The minimum and maximum temperatures reported for growth in laboratory media. 234 . declining in number during storage. 37.8. but not acid conditions. 10.6. with an optimum of 0. parahaemolyticus is easily destroyed by drying.8. 10.FISH AND SEAFOOD Also.9 10.5 pH This organism is tolerant of high pH values.9.4.97 .9.9. 10. with optimum at 7. It can grow in the presence of 0.984 (35).6 Aw/Sodium chloride V.
6 Aw/Sodium chloride V. with a temperature range for growth of 8 . and 2. The optimum concentration of NaCl equates to aw of approximately 0. 41). 10.0.10. respectively. with an optimum pH of 7.10.9. parahaemolyticus will grow at aw levels as low as 0.922.11. its upper and lower limits for growth are 8 (although some reports suggest 10) and 0.43 ºC (42). respectively (39).PATHOGEN PROFILES 10. and are readily destroyed by cooking.8. but the optimum concentration is 2 . it has been reported that cooking oysters for 10 minutes at 50 ºC should ensure destruction of the organism (43).9. vulnificus is heat sensitive. 10.1 Morphology Gram-negative curved rods.2 Oxygen requirements Facultative anaerobe.4% (39.6 .5 minutes.4 Heat resistance V. 10.8 . parahaemolyticus is unable to grow in the absence of salt. 41). 10. parahaemolyticus are not heat resistant. 235 .29 minutes.10 Vibrio vulnificus 10.10. V.4 Heat resistance V.9.02 .98 (35. The D-values at 55 ºC for clam and crab homogenate are 0. 10.6 (41).10.5 pH The pH range for growth is 4.3 Temperature The optimum temperature for growth has been reported at 37 ºC. The organism can survive for long periods at freezer temperatures (42).5% NaCl. 10. depending on solute.
R. in Foodborne Bacterial Pathogens. Cliver D. Hauschild A.H.L..W. 10. 279-303. in Foodborne Disease Handbook.K.W. Eds.O. Marcel Dekker Inc. 1994. References International Commission on Microbiological Specifications for Foods.M. Ed. Aeromonas hydrophila. 2. ASM Press..H. Bier J. Abeyta C. Clostridium botulinum. New York. Austin J. Doyle M. New York. Clostridium botulinum. 1993.N.. with an optimum pH of 7.. Doyle M.10. 3. Fratamico P. Peck M. 1-19. 1993.D. Marcel Dekker. Murrell K.. Ed. New York. Beuchat L.R. Aeromonas.. in Clostridium botulinum: Ecology and Control in Foods. 329-49. Smith J. New York. Lund B.D. Eds. 2001. Marcel Dekker.H. Murrell K. Doyle M. Diseases Caused by Bacteria. in Clostridium botulinum: Ecology and Control in Foods. 10.D. 2003. 1995.H.H. in International Handbook of Foodborne Pathogens.. Montville T.5 pH The pH range for growth is 5 .. Betts G.. 7.R. 11. 1994. Hauschild A. Gorham J.W. London.N. Dodds K. Bhunia A..L.. 5. Dodds K. Novak J. Great Britain. Gorman J. 57-63. Ed. 1989.6 Aw/Sodium chloride V.. Volume 1: Diseases Caused by Bacteria. vulnificus grows over the range 0.0% NaCl.H.... Johnson E. Stelma G. in Foodborne Bacterial Pathogens.5 . New York. Foegeding P.A.E.5.P. Hauschild A. in Microorganisms in Foods.W. Marcel Dekker. 1989.O..8 (42). 97-131. 6.FISH AND SEAFOOD 10. Gaze J. Potential hazards associated with REPFEDS. Marcel Dekker. 111-89.. 121-76.D. 2005. 10. Eds. Marcel Dekker. Eds.M. 357-67.. with an optimum of 2. Eds. in Foodborne Pathogens. Palumbo S. Eds. 5-19. Faris A. Volume 5: Microbiological Specifications of Food Pathogens.10. International Commission on Microbiological Specifications for Foods.W. 1996. Marcel Dekker. Growth and heat resistance of psychrotrophic Clostridium botulinum in relation to ‘sous vide’. Notermans S. 4. Aeromonas hydrophila group.J.10. Aeromonas species. in Food Microbiology: Fundamentals and Frontiers. Caister Academic Press. 6 (1). Kim J.L. Eds.P. Volume 1. Food Control.M. Hui Y. 236 .. 9. Cliver D. Miliotis M.P. New York. Principles of control. Juneja V. Washington D. Clostridium botulinum and Clostridium perfringens. Blackie.. 8.5% (42). Clostridium botulinum. Dodds K. 1-27. New York. Microbiology and Molecular Biology. Hui Y.... in Foodborne Disease Handbook.C. 383-408. Stelma G.11 1.L. Kovacek K.
London. Matthews K.R. 191-243. Varnam A.O. Plesiomonas shigelloides. Growth of Listeria monocytogenes at refrigeration temperatures. Marcel Dekker. McClane B. London. London.D. 311-25. 23. Diseases Caused by Bacteria. 33. Varnam A. Listeriosis and Food Safety. London.O. Labbe R. International Commission on Microbiological Specifications for Foods. Volume 5: Microbiological Specifications of Food Pathogens. London. ASM Press. in Foodborne Bacterial Pathogens. in Foodborne Disease Handbook. 16. 18. Murrell K.R.G. New York. 1990. Wolfe Publishing Ltd. Doyle M.. 351-82. Blackie.A. Juneja V. 13.. 2007. in Microorganisms in Foods. Ed.M. Academic Press.An Introduction. Marcel Dekker..G. Doyle M. Cliver D..H. in Foodborne Pathogens: An Illustrated Text.. Kim C. 15. Labbe R. Doyle M. Rinke H.. Washington D. Ryser E.J.V. Plesiomonas shigelloides.J. 1996. 1990.H. International Commission on Microbiological Specifications for Foods. Koburger J. Wrigley D. 1996. 201-8. Volume 5: Microbiological Specifications of Food Pathogens.G. Eds. Foodborne disease due to Bacillus and Clostridium species. Johnson E.P. Marth E. 133-67.. 1989. Clostridium perfringens.C. 735-42. Washington D. in Foodborne Diseases. in Microorganisms in Foods.P. Evans M. Riemann H. Montville T. Marcel Dekker.P. in Foodborne Infection and Intoxication. 1991.. 229-40. Cliver D. 21.. Clostridium perfringens. Walker S.J. in Foodborne Bacterial Pathogens. 2006. Ed. Ed.. 157-62. Archer P. Journal of Applied Bacteriology.M. 25.PATHOGEN PROFILES 12. Cliver D.K. Emery D. ASM Press. International Commission on Microbiological Specifications for Foods. in Microorganisms in Foods.O. Eds.. 141-82. Eds.A. 129-38. 1990. 237 . New York. Blackie.A. 336 (8721). Eds. Clostridium perfringens. Effect of time and temperature on growth of Salmonella enteritidis in experimentally inoculated eggs. Volume 1... Blackie. 1994. 112-25. 2005. Eds. Lund B. London.T.J. Listeria monocytogenes.A. 1989.. Listeria. Avian Disease. New York.. 14.R. 68 (2).H. 24. Ed.A. 22.. Clostridium perfringens gastroenteritis. Evans M. 1989. 17. Plesiomonas. Clostridium perfringens. International Commission on Microbiological Specifications for Foods. Beuchat L..H. Nagaraja K. 20. in Food Microbiology: Fundamentals and Frontiers. 26. 2001. 19. Hui Y. Volume 5: Microbiological Specifications of Food Pathogens. Elsevier. Montville T. 137-64. Marcel Dekker. Lancet. Ed. 208-13. Listeria monocytogenes. 982-6. Halvorson D.P. in Food Microbiology . New York. Banks J. Clostridium perfringens food poisoning. International Commission on Microbiological Specifications for Foods. Ed. 1996..C. Gorham J. International Commission on Microbiological Specifications for Foods.
Marcel Dekker. 273-84. Marcel Dekker. in Foodborne Pathogens: Microbiology and Molecular Biology. Loessner M. in Understanding Pathogen Behaviour Virulence.S..M.D. 36. Staphylococcus aureus as a food pathogen: staphylococcal enterotoxins and stress response systems. Microbiological Specifications of Food Pathogens. 217-64. Academic Press. 426-35. Eds. Blackie. Blackie. 33. London. Eds. New York. Cambridge. 34.O. Volume 5. R15764.R. 85-106.FISH AND SEAFOOD 27.62.C. in Microorganisms in Foods. Vibrio parahaemolyticus. London. Bergdoll M. Experimental investigations concerning the prevention of epidemic diseases due to aquatic products (Fourth Report). International Commission on Microbiological Specifications for Foods.A. International Commission on Microbiological Specifications for Foods. 1993. 2005. Jay J. 2005. International Commission on Microbiological Specifications for Foods. Staphylococcus aureus. 1989. 1983. Jay J.. 1996. Ed.M. 327-52. 1990. Doyle M. 45 (3). 32. CDR Review. On the resisting power of Vibrio cholerae and typhoid Bacillus against low temperatures. 1996. Vibrio cholerae. International Commission on Microbiological Specifications for Foods..J. Reed G. Wymondham. 414-25. Griffiths M. 4 (13).. Vibrio parahaemolyticus. 1996. in Foodborne Infections and Intoxications. in Foodborne Disease.J. Microbiological Specifications of Food Pathogens. 38. Oliver J.S. Ed. Cliver D. Foodborne illness (Part 1): Staphylococcal (“Staph”) food poisoning. 40. 30.. Loessner M. D’Aoust J-Y. 545-66. 543-68. London. Woodhead Publishing Ltd. Applied and Environmental Microbiology. Blackie. 2005. Twedt R. Smith J. 238 . Salmonella. Warner R. 1994.. Cholera: current epidemiology. Ed. Lee-Wong A. 28.M..P. 331-57. Salmonellae. Ed. Golden D. Ed. Doyle M. 2005.P.A. Gustafson J. 985-98. Food and Environmental Sanitation. Staphylococcal Food Poisoning. International Commission on Microbiological Specifications for Foods. Staphylococcal intoxications. Springer Science.. 642.. Stewart G. Cleland D.C. in Microorganisms in Foods. 1989.. 37.O.P. Fratamico P. in Microorganisms in Foods. Bergdoll M. Wilkinson B. Distribution of Vibrio vulnificus and other lactose-fermenting vibrios in the marine environment.M. New York. Cliver D. International Commission on Microbiological Specifications for Foods. in Foodborne Pathogens. Staphylococcal gastroenteritis. Volume 5. Eds. Japanese Journal of Experimental Medicine. Crowcroft N. London. Ed.H. Academic Press. in Foodborne Bacterial Pathogens. Tohyama Y.S. Stress Response and Resistance. 29. Riemann H. 31. Bhunia A. London. 1930. Caister Academic Press.A. 8. 39. 523. in Modern Food Microbiology. Dairy.L. Golden D. Volume 5: Microbiological Specifications of Food Pathogens. Ed. 13 (11). 327-445. New York. 35.K.
43. Golden D. Ed. 1996.D. 436-9. in Microorganisms in Foods. in Modern Food Microbiology. 2005. Journal of Food Protection.. 1992..W. Loessner M. Foodborne gastroenteritis caused by Vibrio.PATHOGEN PROFILES 41. 657-78.. Cold storage and mild heat treatment as processing aids to reduce the numbers of Vibrio vulnificus in raw oysters. International Commission on Microbiological Specifications for Foods.M.J. Loessner M.A. Blackie. International Commission on Microbiological Specifications for Foods. Yersinia. 985-9.M.A.. Golden D. Volume 5. New York. Jay J. Microbiological Specifications of Food Pathogens.. Jay J. Cook D. London. Ruple A. Vibrio vulnificus. 42. 239 .J. and Campylobacter species. Springer Science. 55 (12).
FISH AND SEAFOOD 240 .
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.co.CONTACTS Addresses of Trade Associations and Professional Bodies Association of British Salted Fish Curers and Exporters c/o Cawoods (Fish Curers) Ltd Estate Rd No 6 South Humberside Industrial Estate Grimsby North East Lincolnshire DN31 2TG United Kingdom Tel: + 44 (0) 1472 342248 Fax: + 44 (0) 1472 353100 Email: firstname.lastname@example.org Web site: www.uk Institute of Fisheries Management (IFM) 22 Rushworth Avenue West Bridgford Nottingham NG2 7LF United Kingdom Tel: + 44 (0)115 9822317 Fax: + 44 (0)115 9826150 Email: email@example.com Prepared Fish Products Association (PFPA) Federation House 6 Catherine Street London WC2B 5JJ United Kingdom Tel: + 44 (0) 208 78362460 The Scottish Government Fisheries Group Environment and Rural Affairs Department Pentland House 47 Robb's Loan Edinburgh EH14 1TY United Kingdom Tel: + 44 (0) 8457 741741 or + 44 (0)131 5568400 Fax: + 44 (0)1397 795001 Email: firstname.lastname@example.org or v.gov.defra.gsi.ifm.uk/Topics/Fisheries Seafish Industry Authority 18 Logie Mill Logie Green Road Edinburgh EH7 4HS United Kingdom Tel: + 44 (0)131 5583331 Fax: + 44 (0)131 5581442 Web site: email@example.com Web site: www.org/ .seafish.
Andersens Boulevard 37.uk/index/fisheriesfarming-and-food.dk/s Norwegian Institute of Fisheries and Aquaculture Ltd Muninbakken 9 .uk Web site: www.gov.13 Postbox 6122 N-9291 Tromsø Norway Tel: + 47 77 629000 Fax: + 47 77 629100 Email: post@fiskeriforskning.C.com/ Danish Institute for Fisheries Technology and Aquaculture (DIFTA) illemoesvej The North Sea Centre DK-9850 Hirtshals Denmark Tel: + 45 98 944300 Fax: + 45 98 942226 Email: firstname.lastname@example.org Web site: www. Baelskaai 25 8400 Oostende Belgium Tel: + 32 59321876 Fax: + 32 59322840 Email: info@eapo. Dublin Eire Ireland Tel: + 353 1 2144100 Web site: email@example.com/ 242 .bim.dk Danmarks Fiskehandlere H.difta.gov.net Irish Sea Fisheries Board PO Box 12 Crofton Road Dun Laoghaire Co.ie/templates/homepage. 1 1553 København V Denmark Tel: + 45 35 372023 Fax: + 45 35 372033 Email: Info@fiskehandlerne.org Web site: www.suite.org/fishery/ European Association of Fish Producers’ Organisations H.fiskeriforskning.matis.no/ Matís ohf (Icelandic Fisheries Laboratories) Skúlagata 4 101 Reykjavík Iceland Tel: + 354 422 5000 Fax: + 354 422 5001 Email: firstname.lastname@example.org Web site: www.eapo.com Web site: www.is Web site: www.asp Food and Agriculture Organization of the United Nations Fisheries and Aquaculture Department Viale delle Terme di Caracalla 00153 Rome Italy Tel: + 39 (06) 57051 Fax: + 39 (06) 57053152 Email: FAO-HQ@fao.no Web site: www. Northern Ireland Fisheries and Rural Policy Division 4th Floor North Dundonald House Stormont Estate Belfast BT4 3SB United Kingdom Tel: + 44 (0) 28 90524999/ 4991 Fax: + 44 (0) 28 90378323 Email: dardhelpline@dardni.FISH AND SEAFOOD Department of Agriculture and Rural Development.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.
au New Zealand Seafood Industry Council Private Bag 24-901 Wellington 6142 New Zealand Tel: + 64 4 3854005 Fax: + 64 4 3852727 Email: email@example.com Web site: www.org.co.bsi-global.gov.ifr.cmar.org.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: firstname.lastname@example.org/ 243 .htm Food and Drink Federation (FDF) 6 Catherine Street London WC2B SJJ United Kingdom TeI: + 44 (0) 207 8362460 Fax: + 44 (0) 207 8360580 Email: email@example.com Web site: www.seafood.hpa.dh.org CSIRO Marine Laboratories Castray Esplanade GPO Box 1538 Hobart Tasmania 7001 Australia Tel: + 61 3 62325222 Fax: + 61 3 62325000 Email: firstname.lastname@example.org Institute of Food Science and Technology (IFST) 5 Cambridge Court 210 Shepherds Bush Road London W6 7NL United Kingdom TeI: + 44 (0) 207 6036316 Fax: + 44 (0) 207 6029936 Email: email@example.com/sc-home Health Protection Agency (HPA) 61 Colindale Avenue London NW9 5EQ United Kingdom TeI: + 44 (0) 208 2004400 Fax: + 44 (0) 208 2007868 Web site: www.CONTACTS Fishing Industry Research Institute 15 Lower Hope Street Rosebank 7700 Republic of South Africa National Fisheries Institute (USA) 7918 Jones Branch Drive Suite 700 McLean VA 22102 United States of America Tel: + 1 703 7528880 Email: firstname.lastname@example.org Department of Health Richmond House 79 Whitehall London SW1A 2NS United Kingdom Tel: + 44 (0) 207 2104850 Email: email@example.com/en/index.au Web site: www.co.gov.ifst.uk 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.org.csiro.com Web site: www.
int/ SIK .asp Codex Alimentarius Commission (CAC) Viale delle Terme di Caracalla 00153 Rome Italy Tel: + 39 (06) 57051 Fax: + 39 (06) 57054593 Email: Codex@fao.int/foodsafety World Health Organization (WHO) Headquarters CH . jsp Food Safety 10 .nhs.se Web site: www.int.net/web/indexen.uk Web site: www.The Swedish Institute for Food and Biotechnology Box 5401 SE-402 29 Göteborg Sweden Tel: + 46 31 3355600 Fax: + 46 31 833782 Email: firstname.lastname@example.org/index.who.euro.scot. Web site: www.sfam.euro.uk/scieh.who.de 244 .scot.int/en/ WHO Regional Office for Europe Scherfigsvej 8 DK-2100 Copenhagen Ø Denmark Tel: + 45 39 171717 Fax: + 45 39 171818 Email: email@example.com/ Health Protection Scotland Clifton House Clifton Place Glasgow G3 7LN United Kingdom Tel: + 44 (0)141 3001100 Fax: + 44 (0)141 3001170 Email: firstname.lastname@example.org Rome Italy Tel: + 39 06 487751 Fax: + 39 06 4877599 Email: email@example.com Web site: www.who.who.FISH AND SEAFOOD Society for Applied Microbiology Bedford Heights Brickhill Drive Bedford MK41 7PH United Kingdom Tel: + 44 (0) 1234 326661 Fax: + 44 (0) 1234 326678 Web site: www.php Society of Food Hygiene and Technology The Granary Middleton House Farm Tamworth Road Middleton Staffs B78 2BD United Kingdom Tel: + 44 (0) 1827 872500 Fax: + 44 (0) 1827 875800 Email: firstname.lastname@example.org/ FAO/WHO Collaborating Centre for Research and Training in Food Hygiene and Zoonoses Robert von Ostertag Institute Postfach 330013 D-14191 Berlin Germany Tel: + 49 (0) 188 84122155/ 2157 Fax: + 49 (0) 188 84122957 Email: email@example.com Geneva 27 Switzerland Tel: + 41 22 7912111 Fax: + 41 22 7913111 Email: firstname.lastname@example.org Web site: http://www.org Web site: www.int Web site: www.codexalimentarius.
ift.FoodSafety.sofht.cdc.CONTACTS US FDA .int/foodsafety/en/ European Commission: Activities of the European Union food safety http://europa.org Web site: http://www.fda.eu/pol/food/index_en. Beverage and Consumer Products Companies http://www.org/ Royal Institute of Public Health (UK) http://www.foodscience.org/ Institute of Food Technologists http://www.foodprotection.gov/ Institute of Food Technologists 525 W.Center for Food Safety and Applied Nutrition 5100 Paint Branch Parkway College Park MD 20740-3835 United States of America Tel: + 1 888 SAFEFOOD (+ 1 888 723 3366) Web site: www.org/cms/ Useful Web Sites Gateway to Government food safety information (US) http://www.co.uk/ Society of Food Hygiene and Technology (UK) http://www.fsai.who. Van Buren Suite 1000 Chicago IL 60607 United States of America Tel: + 1 3127828424 or + 1 8004383663 Fax: + 1 3127828348 Email: email@example.com/ The Association of Food.org.htm Centre for Disease Control and Prevention (CDC) (US) http://www.gov/foodsafety/ Food Safety Authority of Ireland http://www.cfsan.fpa-food.gov/ World Health Organization (WHO): Food safety programmes and projects http://www.ie/ Food Science Australia http://www.uk/ 245 .riph.ift.csiro.au/ International Association for Food Protection http://www.
FISH AND SEAFOOD 246 .
as autochthonous bacterial flora of fish 3 Aeromonas spp. 102 Bacillus spp. source of PSP toxins 142 Alexandrium ostenfeldii. as spoilage organism in oysters 65 as spoilage organism in tropical crustacean shellfish 81 in molluscan shellfish 60 Alexandrium. in crustacean shellfish 86-7 ATP. growth and survival in crustacean shellfish 86 Aeromonas sobria. as TTX producers 145 Amnesic shellfish poisoning (ASP) toxins 146-8 caused by domoic acid in crustacean shellfish 87 Anchovies. 69-70 Aeromonas hydrophila. in crustacean shellfish 87 Alteromonas spp. as autochthonous bacterial flora of fish 3 as spoilage organisms in oysters 65 as spoilage organisms in temperate crustacean shellfish 80 as spoilage organisms in tropical crustacean shellfish 81 in molluscan shellfish 60 Aerobic plate counts. growth and survival in crustacean shellfish 86 Air drying 101 Alcaligenes. as autochthonous bacterial flora of fish 3 in fish sauce 127 Bacillus cereus.INDEX Achromobacter spp. growth and survival in chilled and frozen raw fish 19 of concern in preserved fish 115 Aspergillus spp. causing spoilage of dried fish 110. to test microbiological criteria 19 Aeromonadaceae. breakdown of – as indicator of fish freshness 10-1 Azaspiracids 158-9 detection methods 160 origins and distribution 159 toxicology 159-60 Bacalao 97. preserved by dry/wet salting 99 Anisakis spp. 111 Astroviruses. 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 . planktonic source of spirolides in shellfish 163 Allergens. 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.
caused by Vibrio cholerae O1 17 Ciguatoxins 155-8 detection methods 157-8 origins and distribution 1567 toxicology 157 Clams. mould causing spoilage of dried fish 110 Battering process 36-8 Bivalves. 226 Breading process 36-8 Brevetoxins. as autochthonous bacterial flora of fish 3 Clostridium botulinum. 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. 113. associated with consumption of fish/fish products 16. concern with respect to use of MAP 7 growth and survival in chilled and frozen prepared fish products 45 growth and survival in chilled and frozen raw fish 15-6 growth and survival in crustacean shellfish 84 growth and survival in cured. of fish – effects on microflora 5-6 Carbohydrate. fish sauce 123 Basipetospora halophila. in crustacean shellfish 86-7 Campylobacter coli. growth and survival in crustacean shellfish 86 growth and survival in molluscan shellfish 59-60. affected by azaspiracids 159 affected by cyclic imines 161 Botulism. as gas in modifiedatmosphere packaging 6 CCP Decision Tree (in HACCP plan) (Fig) 185 Ceviche. of fresh fish spoilage 11 Chemical pollutants. 136. 137.FISH AND SEAFOOD components of seafood ready meals 40 Bacterial pathogens. for PSP toxins 144 Chemical indicators. importance in lactic acid fermentation in fish products 130. description and uses 56-7 effects of processing on microflora 60-4 hot-smoking 107 Clear smoke treatments 108 Clostridium. 68-9 Capture and handling. 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. as cause of spoilage of MAP fish 15 Calciviruses. 131 Carbon dioxide. 98-9 Brochothrix thermosphacta. indigenous – in fish (Table) 4 non-indigenous – in fish (Table) 4 Bakasang. preserved by marinating 100 Chemical detection methods. 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 .
149-50. to determine fresh fish spoilage 11 Eels. as autochthonous bacterial flora of fish 3 Crabs. smoked and dried fish 114 pathogenic microorganisms in molluscan shellfish 59-60 249 . of fish – analysis of 10 Diarrhetic shellfish poisoning (DSP) toxins 148-51 detection methods 150-1 origins and distribution 14950 toxicology 150 Dinophysis spp. to detect NSP toxins 155 ELISA kit (Biosense).INDEX pathogen profile 226-7 Clostridium perfringens. growth and survival in chilled and frozen raw fish 19 of concern in cold-smoked salmon 115 Domoic acid (DA). smoked and dried fish 114 Enzymic spoilage. hot-smoking 107 ELISA assay. of fish – different processes 94 Dun. cooked – effects of processing on microflora 83 forms for purchase 80 freshness indicators 83 Crayfish. forms for purchase 80 Crustacean shellfish 79-92 Cured fish 93-121 Cyclic imines 160-1 detection methods 162 origins and distribution 161 toxicology 162 Degradation. as 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. 152 Diphyllobothrium. 106 Contacts 241-5 Conway method. as contaminants in scampi 36. adverse condition on poorly preserved fish 110 EC grading scheme. growth and survival in crabmeat 84 growth and survival in cured. of prepared fish products 44 Escherichia coli. for detection of DA 148 ELISA kit (DSP-Check). 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. description and uses 59 effects of processing on microflora 60-4 Cold-smoking 104-5. 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. as spoilage organisms in tropical crustacean shellfish 81 Coryneforms. to detect DSP toxins 150-1 Enrobing process 36-8 Enterobacteriaceae. to determine fresh fish spoilage 11 Corynebacteria. source of azaspiracids 159 source of DSP toxins 148-9. processing 334 Cockles.
fermented fish product 124 General Pathogen Surveillance Programme 84 Gravad fish. preserved by marinating 100 preserved by dry/wet salting 99 Histamine fish poisoning. 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. in crustacean shellfish 86-7 in molluscan shellfish 59-60 Hepatitis A virus. source of PSP toxins 142 Gyoshoyu.FISH AND SEAFOOD EU legislation. concerning crustacean shellfish 87-8 European Grading Scheme. of fresh fish spoilage 911 Evisceration. 130-5 Fish-spoilage bacteria 12 Fishtester. as autochthonous bacterial flora of fish 3 in molluscan shellfish 60 spoilage organism in oysters 65 Food hygiene legislation 189-224 Food of animal origin. as source of ciguatoxins 156 Garlic. fermented fish product 126 Green cured split cod 97. specific EU hygiene rules 196-224 Fresh fish. of fish – effects on microflora 5-6 Hepatitis (non-A. for evaluation of fish quality 9 Eurotium. caused by Photobacterium phosphoreum 15 Hon kare bushi 107 Hot-smoking 105-7 250 . 102 Gymnodimines. preserved by marinating 101 Gravlaks. definition of terms 176-7 implementation and review of plan 184-6 in fish and seafood product manufacture 175-88 seven stages of system 17784 Halobacterium spp. of prepared fish products 44 Gambiertoxins. in crustacean shellfish 86-7 in molluscan shellfish 59-60 Herring. definition 1 Fermented fish 123-40 Fermented fish/salt products of Thailand (Table) 125 Fish and shellfish toxins 141-74 Fish cakes. spoilage 9-11 Freshness grading schemes 9 Frozen prepared fish products 27-51 Frozen raw fish 1-25 Frozen storage. processing 35 Fish component. mould causing spoilage of dried fish 110 Evaluation. causing pink condition on fish surface 110 in fish sauce 127 Halococcus spp. of fish at sea 5 Fatty fish. non-B) viruses. effects of processing 127-30 Fish sauces and pastes of south-east Asia (Table) 124 Fish/salt/carbohydrate products of Asia (Table) 125. member of cyclic imine group 161 Gymnodinium catenatum. fish sauce 123 HACCP. as source of inulin in lactic acid fermentation in fish products 134 Garos. to measure fish skin resistance 10 Flavobacterium.
in fish fillet 1 Lipid oxidation. initial – of chilled and frozen prepared fish products 31-2 of chilled and frozen raw fish 3-5 251 . growth and survival in chilled and frozen prepared fish products 46 growth and survival in cured. enabled by carbohydrate source in fish products 130. outbreak caused by gravid 115 Lobster. smoked and dried fish (Table) 116-7 for shellfish products 70 Microbiological limits. as source of yessotoxins 152 Lipid content. preserved by marinating 101 Microbiological criteria. for chilled and frozen prepared fish products 47-9 for chilled and frozen raw fish 19-20 for crustacean shellfish 87-8 for cured. cooked – effects of processing on microflora 83 forms for purchase 80 freshness indicators 83 Mackerel. leading to oxidative rancidity 11 Lipophilic shellfish poisoning toxins 149.INDEX Hygiene regulations for foodstuffs production and distribution (EU) 190-6 Ice storage. 131 Lactic fermented fish products (Table) 126 fish species used in the Philippines (Table) 126 Lactobacillus. hot-smoking 107 preserved by dry/wet salting 99 Marinating 94. 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. as autochthonous bacterial flora of fish 3 Lean fish. as source of botulism 136 International legislation. producer of brevetoxins 154 Katsuobushi 107 Kench curing 97-8 Klipfish 97 Lactic acid bacteria. smoked and dried fish 114-5 hazard in cold-smoking 105 hazard in mild-processed marinated products 101 Listeriosis. concerning crustacean shellfis 88 Jellied eels. causing spoilage in vacuum-packed dried salmon 111 Lactic acid fermentation. for microbial inactivation in fish products 127. preserved by salt-boiling 100 Ionising radiation. 100-1 Masmeen 107 Matjes. as hazard in regard to reef-caught fish 28 growth and survival in crustacean shellfish 84 pathogen profile 229-30 Listeria monocytogenes. definition 1 Legislation. to reduce rate of fish spoilage 12 Ikan kaju. food hygiene 189-224 Lingulodinium polyedrum. for fresh and frozen fish (Table) 20 Micrococcus spp. preserved by salt-boiling 100 Karenia brevis. 151-3 detection methods 153 Liquid smoke 107-8 Listeria spp. 135 I-zushi.
reduction in smoking process 108 Norcardiopsis dassonvillei. contaminated by azaspiracids 158 description and uses 58 effects of processing on microflora 60-4 hot-smoking 107 Nam-budu. shigelloides. smoked and dried fish 95 of fermented fish 126-7 of molluscan shellfish 59-60 Microflora. for detection of azaspiracids 160 for detection of ciguatoxins 157 for detection of DA 148 for detection of DSP toxins 150. growth and survival in chilled and frozen raw fish 17 Paralytic shellfish poisoning (PSP) toxins 142-4 detection methods 144 toxicology 143 252 . 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. caused by lipid oxidation 11 Oysters. as autochthonous bacterial flora of fish 3 as spoilage organism in temperate crustacean shellfish 80 in molluscan shellfish 60 Mouse bioassay.FISH AND SEAFOOD of crustacean shellfish 80-1 of cured. in crustacean shellfish 867 Norwalk virus. smoked and dried fish – effects of processing 96-108 of fermented fish – effects of processing 127-35 of fish – effects of processing 5-9 of ingredients of chilled/frozen prepared fish products – effects of processing 32-42 of molluscan shellfish – effects of processing 60-5 Modified-atmosphere packaging 6-9 of fish – (US) NACMCF recommendations 7-8 of fish – Codex Committee guidance 7 of fresh chilled products 42 Molluscan shellfish 53-77 commercial products 54-9 definitions 53-9 Moraxella. fish sauce 123 Neosaxitoxin (neoSTS) 142-3 Neurotoxin shellfish poisoning (NSP) toxins 153-5 detection methods 155 Nitrosamines. associated with consumption of fermented fish products 137 Oxidative rancidity. of crustacean shellfish – effects of processing 81-3 of cured. as TTX producer 145 Norovirus. contamination leading to illness 68 description and uses 57 effects of processing on microflora 60-4 hot-smoking 107 spoilage organisms 65 P.
69-70 Polycyclic aromatic hydrocarbons. as source of DSP toxins 148-9 Protein content. reduction in smoking process 103. as component for seafood ready meals 40 Pathogen profiles. as source of toxins 159. detection methods 165 origins and distribution 165 toxicology 165 Pissala. causing spoilage of dried fish 110-1 Photobacterium phosphoreum. 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. 225-39 Pathogens. adverse condition on poorly preserved fish 110 Pinnatoxins member of cyclic imine group 164. pathogen profile 230 in molluscan shellfish 5960. growth and survival in chilled and frozen raw fish 19 growth and survival in crustacean shellfish 86 growth and survival in cured. smoked and dried fish 115 Pasta. as source of yessotoxins 152 Protoperidinium cassipes. 81 as spoilage organism in fresh fish 13 as spoilage organism in oysters 65 as TTX producers 145 253 . as component for seafood ready meals 40 Prawns. 161. effects of processing on microflora 81-2 freshness indicators 83 Pre-column derivatisation. 163. planktonic source of azaspiracids 159 Pseudomonadaceae. as autochthonous bacterial flora of fish 3 as spoilage organism in chilled products 43 as spoilage organism in crustacean shellfish 80. mould causing spoilage of dried fish 110 Potato. spoilage organism in oysters 65 Protoceratium reticulatum. for detection of PSP toxins 144. 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. in molluscan shellfish 60 Pseudomonas spp. 197 Plesiomonas. effects on microflora of fish 5-9 Professional bodies. in fish fillet 1 Proteus. 145-6 definitions 28-30 non-seafood ingredients 32 Processing. 152-3 Penicillium spp. 100-1 Pindang. preserved by salt-boiling 100 Pink. smoked and dried fish 113-6 growth and survival in fermented fish 136-7 growth and survival in molluscan shellfish 66-70 Pectenotoxins 151. 108 Polypaecilum pisce.INDEX Parasites. causing spoilage in vacuum-packed dried fish 111 Pseudomonads.
in molluscan shellfish 60 Rice. preserved in saturated brine 99 Sauce. smoked and dried fish 114 pathogen profile 230-1 pathogenic microorganisms in molluscan shellfish 59-60 Salt burn. processing 34. detected in cod 115 Pteriatoxins. resulting from poorly salted cod 110 Pyrodinium. for microbial inactivation in fish products 127. initial microflora 31 Seafood ready meals 39-40 Seafood recipe meal centres 38-9 Shaped fish products. vacuum packing 105 Salmonella spp. for fish ready meals – manufacture 40-2 Saxitoxin (STX) 142-3 Scallops. importance in lactic acid fermentation in fish products 130 importance in salting.) 41 Recipe fish products 38-40 Rhodotorula rubra. as spoilage organism in chilled products 43 as spoilage organism in fresh fish 13-4 as spoilage organism in temperate crustacean shellfish 80. processing 35 Shellstock.FISH AND SEAFOOD inhibition of growth by carbon dioxide 6 Pseudonitzschia multiseries. to measure fish skin resistance 10 Salmon. flow diagram (Fig. antibody-based kit to detect STX 144 RT Freshmeter. detection methods 165 member of cyclic imine group 164 origins and distribution 165 toxicology 165 Puffer fish. 135 Rakefisk. source of PSP toxins 142 Quality Index Method. assembly and packaging. chilled and frozen 1-25 Recipe dish. as hazard in regard to reef-caught fish 28 growth and survival in crustacean shellfish 84 growth and survival in cured. as autochthonous bacterial flora of fish 3 inhibition of growth by carbon dioxide 6 Shewanella putrefaciens. fermented fish product 126 as source of botulism 136 Rakørret. 100 Salting 97-9 definitions 93-4 Sardines. implicated in amnesic shellfish poisoning 146-7 Pseudoterranova. fermented fish product 126 Raphidophytes. producers of brevetoxins 154 Raw fish. 36 Seafood. for evaluation of fish quality 9 Radiation. 83 254 . hazard in salting of fish 97 Salt concentration. description and uses 58-9 Scampi. flow diagram (Fig. as carbohydrate source in lactic acid fermentation in fish products 134 as component for seafood ready meals 40 RIDASCREEN.) 41 ingredients preparation. growth and survival in chilled and frozen raw fish 19 Pseudoterranova dicipiens. smoking and marinating of fish 96 Salt-boiling 94. as source of tetrotoxin 144-5 Putty fish. processing and effects on microflora 60-1 Shewanella.
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. detection methods 164 member of cyclic imine group 162-4 origins and distribution 163 toxicology 163-5 Spoilage. pathogenic microorganisms in molluscan shellfish 59-60 Specific spoilage organisms (SSOs) 12-5 Spirolides. role in fish degradation 10. fermented fish product 126 Taste panel evaluation. as measure of fish degradation 10 Sliming. growth and survival in chilled and frozen prepared fish products 46 growth and survival in cured. definitions of forms for purchase 79-80 effects of processing on microflora 81-2 freshness indicators 83 Shucked shellfish. of fish after capture 5 Surstrømming. 101 Storage. smoked and dried fish 114 pathogenic microorganisms in molluscan shellfish 59-60 Shottsuru. of crustacean shellfish 80. fish and shellfish 141-74 Trade associations. contact details 241-2 Trematode infections foodborne 137 Trichosporon. of cooked fish 9-10 Tetrotoxin 144-6 detection methods 145-6 toxicology 145 TMAO.INDEX Shigella spp. as indicator of fresh fish spoilage 11 Toxins. to measure fish skin resistance 10 Total Volatile Base-Nitrogen. fish sauce 123 Shrimps. 12-5 of recipe fish meals 43-4 of shellfish 65 Staphylococcus. grading of 31 hot-smoking 107 US legislation. concerning crustacean shellfish 89 Vacuum packaging. processing and effects on microflora 63-4 Skin resistance. in molluscan shellfish 60 Tuna. 13. 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. 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. for evaluation of fish quality 9 Torrymeter. 83 of fermented fish 135-6 of fish subjected to different preservation methods 108-10 of fresh fish 9-11. 14 Torry Scale.
causing dun condition on dried fish 110 Wet salting 98-9 Wet storage. growth and survival in crustacean shellfish 85 pathogen profile 233-4 Vibrio cholerae 01.FISH AND SEAFOOD guidance 7 of fresh chilled products 42 Vegetables. as hazard in regard to reef-caught fish 28 as spoilage organism in crustacean shellfish 80. processing for seafood recipe meal centres 39 Vibrio spp. growth and survival in chilled and frozen raw fish 16-7 growth and survival in crustacean shellfish 86 growth and survival in molluscan shellfish 67 pathogen profile 235-6 Vibrionaceae. as spoilage organisms in chilled recipe meals 43 Yessotoxins 151. 81 as TTX producers 145 pathogenic microorganisms in molluscan shellfish 59-60 risk in cold-smoking 105 Vibrio cholerae. growth and survival in chilled and frozen raw fish 17 Vibrio cholerae 01 and 0139. 152-3 256 . growth and survival in chilled and frozen raw fish 16-7 growth and survival in crustacean shellfish 86 growth and survival in fermented fish 136 growth and survival in molluscan shellfish 66-67 pathogen profile 234-5 Vibrio vulnificus. of shellstock 61. growth and survival in chilled and frozen prepared fish products 47 growth and survival in crustacean shellfish 86-7 Wallemia sebi. 68 Vibrio cholerae non-01. growth and survival in molluscan shellfish 67-8 Vibrio parahaemolyticus. as autochthonous bacterial flora of fish 3 as spoilage organisms in chilled products 43 as spoilage organisms in vacuum-packed dried fish 111 Viruses. 62 Yersinia spp.