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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
I also acknowledge and thank Jackie Apps. Alison Turner. Acknowledgements All the authors involved in writing the first edition of this book are gratefully acknowledged. for his scientific input and Victoria Emerton. The books are intended as a source of information for microbiologists and food scientists working in the food industry and responsible for food safety. and growth characteristics for key pathogens associated with the product. and a detailed list of contacts for various food authorities. thank you for all the support. Training Manager. potential hazards associated with the consumption of a range of products. All handbooks also contain a review of the related legislation in Europe and UK. My gratitude is extended to Dr Peter Wareing. Technical Team Leader for her valuable editorial contribution. and Meat Products. Fish and Seafood. both in the UK and elsewhere. guides to HACCP. and Ann Pernet for the work put into typesetting and indexing. which has been the chief factor in ensuring the completion of this book. published by Leatherhead Food International and RSC Publishing. I dedicate this book to my husband . I thank the contributing authors for their collaboration. The books in the series are designed as easy-to-use guides to the microorganisms found in foods.Goldwyn. Rhea Fernandes Leatherhead Food International v . Each book provides a brief overview of the processing factors that determine the nature and extent of microbial growth and survival in the product.FOREWORD The Microbiology Handbook series includes Dairy Products.
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 2.7 References 3.4 Spoilage 4.CONTENTS CONTRIBUTORS FOREWORD INTRODUCTION 1.3 Processing and its Effects on the Microflora 1.5 Pathogens: Growth and Survival 3.2 Initial Microflora 1.6 Published Microbiological Criteria 3.2 Initial Microflora 3.2 Definitions 2.7 Published Microbiological Criteria 2.5 Factors Affecting Fresh Fish Spoilage 1.8 References 1. 4.4 Spoilage 3.2 Initial Microflora 4. vii .4 Processing and its Effects on the Microflora 2.8 References MOLLUSCAN SHELLFISH 3.1 Introduction 2.1 Definitions 3. 3.9 Further Reading CHILLED AND FROZEN PREPARED FISH PRODUCTS 2.5 Spoilage 2.8 Further Reading CRUSTACEAN SHELLFISH 4.1 Definitions 1.3 Initial Microflora 2.7 References 4.1 Definitions 4.3 Processing and its Effects on the Microflora 3.5 Pathogens: Growth and Survival 4.6 Pathogens: Growth and Survival 2.3 Processing and its Effects on the Microflora 4.4 Spoilage 1.7 Published Microbiological Criteria 1. CHILLED AND FROZEN RAW FISH 1.6 Pathogens: Growth and Survival 1.6 Published Microbiological Criteria 4.
4 Regulation (EC) No.8 Ciguatoxins 7. 8.1 Introduction 7.7 Neurotoxin Shellfish Poisoning (NSP) Toxins 7.3 Processing and its Effects on the Microflora 6.9 Other Relevant Legislation 223 9.I.2 Paralytic Shellfish Poisoning (PSP) Toxins 7.3 Stages of a HACCP Study 8. 854/2004 of the European Parliament and of the Council Laying Down Specific Rules for the Organisation of Official Controls on Products of Animal Origin Intended for Human Consumption 212 9. 9. 852/2004 on the General Hygiene of Foodstuffs 191 9. 2073/2005 on Microbiological Criteria for Foodstuffs 214 9.12 References HACCP IN FISH AND SEAFOOD PRODUCT MANUFACTURE 8.10 Further Reading 223 viii .6 Published Microbiological Criteria 5.6 Regulation (EC) No.7 Food Hygiene (England) Regulations 2006. S. SMOKED AND DRIED FISH 5.5 Diarrhetic Shellfish Poisoning (DSP) Toxins 7.1 Introduction 189 9.1 Definitions 5.11 Conclusion 7. CURED.4 Spoilage 5.7 References 5. 853/2004 Laying Down Specific Hygiene Rules for Food of Animal Origin 196 9.8 Guidance 222 9.10 Cyclic Imines 7.5 Pathogens: Growth and Survival 6.3 Tetrodotoxin 7.7 Further Reading FISH AND SHELLFISH TOXINS 7.8 Further Reading FERMENTED FISH 6.2 Initial Microflora 5.3 Processing and its Effects on the Microflora 5.2 Definitions 8.5 Regulation (EC) No. EU FOOD HYGIENE LEGISLATION 189 9.3 Regulation (EC) No.6 References 6.2 Legislative Structure 190 9. 14 (Hygiene requirements specific to the UK) 221 9.4 Amnesic Shellfish Poisoning (ASP) Toxins 7.5.2 Initial Microflora 6. 7.4 Implementation and Review of the HACCP Plan 8.1 Introduction 8.6 Lipophilic Shellfish Toxins (LST) 7.9 Azaspiracids 7.5 References 93 93 95 96 108 113 116 117 120 123 123 126 127 135 136 137 140 141 141 142 144 146 148 151 153 155 158 160 165 166 175 175 176 177 184 186 6.5 Pathogens: Growth and Survival 5.1 Definitions 6.4 Spoilage 6. 2006 No.
4 Listeria spp.10 PATHOGEN PROFILES 10.6 Salmonella spp.10 Vibrio vulnificus 10.1 Aeromonas spp. 10.5 Plesiomonas 10. 10.11 References 225 225 226 227 229 230 230 232 233 234 235 236 241 247 CONTACTS INDEX ix .3 Clostridium perfringens 10.9 Vibrio parahaemolyticus 10.8 Vibrio cholerae 10.7 Staphylococcus aureus 10.2 Clostridium botulinum 10. 10.
fish and seafood can be involved in the transmission of pathogenic microorganisms and toxins. Meat.. 94767. 2006. cured.. Seafood Research from Fish to Dish: Quality.. Eds. and. Further Reading General Fish/Seafood Microbiology Cross N.INTRODUCTION Fish and seafood are a main source of animal protein in the diet. chilled and frozen prepared fish. Chandan R. season.M. xi . whether they are pelagic (surface to mid-water) or demersal (bottom) feeders will influence the numbers and types of microorganisms present on freshly caught seafood. Poultry. Luten J.A. Often contamination is from human and animal sources. The Microbiology Handbook. Clark S. The second edition of this handbook is a review of the entire book for currency of information. molluscan shellfish. Safety and Processing of Wild and Farmed Fish.. Nollet L. Because of their health advantages over red meats. Wageningen Academic Publishers. in Handbook of Food Products Manufacturing: Health. the consumption of fish and seafood has increased. rivers and lakes whose water can range from pristine to contaminated. smoked and dried fish. Catches can be gathered from seas. Hui Y. crustacean shellfish.. 2007. Seafood processing: basic sanitation practices. for fish. Wiley.Fish and Seafood consists of the microbiology of seven different product categories: chilled and frozen raw fish. Cross N. Saebo A.. Geographical region. Bekaert K.H. Oehlenschlager J. and fish and shellfish toxins. Key changes in this edition are the recent regulatory changes pertaining to food hygiene and microbiological criteria for foodstuffs. Milk.. Seafood and Vegetables. thus.B. fermented fish. Hoboken. Jacobsen C. Wageningen..
J. 2005.D. The fish industry. Gorham J. Martin R. in Minimal Processing Technologies in the Food Industry. Luten J. Seafood processing: basic sanitation practices.. Cambridge. Ed. Blech Z.B. Smith J. Hui Y.R. Blech Z. Hall G. The incidence of Listeria spp. Amsterdam. International Journal of Food Microbiology.. Ohlsson T.. Fish and fish products (Microorganisms in fish products).B. Proceedings of the International Seafood Conference. London. Microbiological spoilage of fish and fish products.International Commission on Microbiological Specifications for Foods. New York. 174-249. November 1995. in Fish Processing Technology. 61 (3). Huss H.L. in Advances in Food and Nutrition Research.. 167-77. New York. 1998. 1997. 1996.H.. 372-6. 2002.. Biogenic amines in fish and shellfish. in Microorganisms in Foods 6: Microbial Ecology of Food Commodities. 200-23... Predictive microbiological modelling and seafood quality. Proceedings of the International Seafood Conference.. Oxford.H. Bengtsson N. Marcel Dekker.. 8 (2).. Woodhead Publishing Ltd. Huss H. 1996.. Eds. Kluwer Academic / Plenum Publishers. Heinitz M. 87121.H. and Clostridium botulinum in smoked fish and shellfish.H. in Seafood from Producer to Consumer: Integrated Approach to Quality. xii . Huss H.L. Gram L. Elsevier. Hafsteinsson H.Y. 543-61. Hui Y. Eds. 1997.. London. Microbiology of fish and fish products.L. in Food Processing: Principles and Applications. 1996. Journal of Food Protection. Noordwijkerhout. 1997. 60 (4). Nip W-K. McCarthy S.E. Ed. Ventresca P. Johnson J. Minimal processing in practice: seafood. 1997. Eds.S. Spoilage and shelf-life extension of fresh fish and shellfish. 1997.K. Bruinsma B. Ed. Vo1ume 39.Y.A. 33 (1). Control of indigenous pathogenic bacteria in seafood. 2002. November 1995. 329-65. 413-30. 91-8. Noordwijkerhout. Gudmundsson M. Seafood: processing. Blackwell Publishing. Ed. 36 (1-2). Dalgaard P. 318-23. Amsterdam.. Blackwell Publishing.P. Cambridge.M.. 2003. Taylor S. 121-37.S. Davies A. Salmonella spp. International Commission on Microbiological Specifications for Foods. Journal of Food Protection. Flick G. Ed.N. 2004. Dalgaard P. Modified-atmosphere packaging of fish and fish products. Ed. Safety and Quality Issues in Fish Processing.R. 459-72. Stanfield P. Luten J.H. Smith J. Incidence and survival of Listeria monocytogenes in ready-to-eat seafood products..A. Academic Press. CRC Critical Reviews in Food Science and Nutrition. Woodhead Publishing Ltd. 245-66 Bremner H. Elsevier. in Seafood from Producer to Consumer: Integrated Approach to Quality. Blackie. Tong P.. 431-43. Rawles D. Gram L. Stanfield P. Food Control.M. Simpson B. basic sanitation practices. in Kosher Food Production.A. Oxford. 2004.. Ashie I. in Food Plant Sanitation.
Chapman and Hall. 25-39. Berlin.R. Huss H. guidelines. Ed. Evaluation of the bacteriological quality of seafood. in Microorganisms in Foods. 2257. Microbiological quality of fishery products. Elsevier. Ward D. Olley J. Microbiological indicators and seafood processing.R. Huss H. Food Technology. Twigg B. in Quality Assurance in the Fish Industry: Proceedings of an International Conference. Jakobsen M. Liston J. Copenhagen.H. Elsevier. 265-79. Eds. International Commission on Microbiological Specifications for Foods. Bonnell A. 16 (1). Bacterial pathogens transmitted by seafood. 1992. 359-403. Food Science and Technology Today. Gilbert R. in Quality Attributes and Their Measurement in Meat. Ross T. Liston J. Sofos J. 1995. London.. Chapman & Hall. Amsterdam. Qualitative and quantitative characterization of spoilage bacteria from packed fish. 1992. August 1991. International Journal of Food Microbiology. 54-61. 1989. Eds. 44 (12). London. Glasgow. Copenhagen. Garrett E. in Public Health Aspects of Seafood-borne Zoonotic Diseases. Elsevier. Fish: microbiological spoilage and safety. Dutson T. 1994. Blackie. xiii . 11 (2). Gibson D. 90-106. Liston J.D. 1996. Application of predictive microbiology to assure the quality and safety of fish and fish products. 118-20. 26 (3).R... August 1991. 1992. Sterling Publications Ltd. in Fisheries Processing: Biotechnological Applications. 1994. Bernoth E-M. Huss H. Poultry and Fish Products. 319-33. in Quality Assurance in the Fish Industry: Proceedings of an International Conference. Ward D. 46 (1). Blackie. Seafood toxins of microbiological origin. Amsterdam.D.. 269-82. Amsterdam. Bacterial spoilage of seafood. 459-78. in Quality Assurance in the Fish Industry: Proceedings of an International Conference.. Gram L. Activities Report of the R&D Associates. PHLS Microbiology Digest. and specifications for seafood products. Evaluation of the bacteriological quality of seafood. 1994.S. Ed. poultry and fish. Martin A. A new look at microbiological standards. August 1991.. International Journal of Food Microbiology. 1994. Copenhagen. 1990. Pearson A. 1-17.International Commission on Microbiological Specifications for Foods. 1991.M. 93-105. Modelling of microbial activity and prediction of shelf-life for packed fish. Ed. London.N. 26 (3). in Quality Assurance in Seafood Processing: A Practical Guide. 305-17.M.J. London. Liston J. Ed. Dalgaard P. Hobbs G.. 1994. Volume 5: Microbiological Specifications of Food Pathogens.H. Bonnell A. Ed. McMeekin T.. Institut fur Veterinarmedizin. 121-8. in Food and Beverage Technology International: USA. Eds.. International Journal of Food Microbiology. Fish spoilage and safety. 5662. Dalgaard P. 166-73. 1990. Microbial growth and its control in meat.H.A. Microbial hazards of seafood consumption. 1995.A. Gram L. Jakobsen M. 1992. 5 (3). The microbiology of seafood products. Liston J.M. Jakobsen M. Eds.
1985. Microbiology of fish. 42 (3). Applied Science Publishers. The bacteriology of fish handling and processing.S.Pedrosa-Menabrito A. 1986.spoilage of fish. 90-3. in Microbiology of Frozen Foods. Hackney C. 1988. Microbiological hazards associated with fishery products. Elsevier Applied Science Publishers.K. Journal of Food Quality. London. 11 (2).R. 71-117.R. Food Technology. Hodgkiss W. Simmonds C. Garrett E. Dicharry A. guidelines.. 1988.M. Davies R. Ed. Volume 1. 1987. Regenstein J. in Essays in Agricultural and Food Microbiology. 169-208. xiv .C. 46 (1). Hobbs G. Shelf-life extension of fresh fish . Robinson R. CSIRO Food Research Quarterly. Seafood-borne bacterial pathogens of marine origin.. Microbiology of frozen fish and related products.. Chichester.J. 199-226. Lamprecht E.K. Hobbs G. in Developments in Food Microbiology. 104-9.. 8-16. 103. Norris J. Wiley. Eyles M. Part I . Microbiological standards. 1988. Food Technology. 42 (3). Ed. and specifications and inspection of seafood products.a review. 1982. Ed. 117-27. London.
1 . Fish fillet composition can vary significantly within the same species due to feed intake. Covadonga Arias Department of Fisheries and Allied Aquacultures Auburn University Auburn Alabama 36849 United States of America 1. it shows a typical season pattern especially in migratory species such as herring or mackerel.21%. This super class of vertebrates includes all the bony and cartilaginous finfish. while fatty fish have fat cells distributed along their bodies. Although fish is broadly similar in composition and structure to meat there are a number of distinctive features.20%. ‘fish’ also refers to the flesh of such animals used as food. Muscle composition and structure of fish also differ from those found in other meat. respectively (1). However. and is mostly interspersed between the muscle fibres. some regulatory agencies such as the US Food and Drug Administration (FDA) will include molluscan shellfish. “fish” will be used for fresh and seawater finfish.5% (1). crustaceans. In addition. most fish contain omega 3-fatty acids and other essential nutrients. and excludes molluscs and Crustacea. The connective tissue content of fish is also lower than that found in meat. and spawning season. Protein content in fish fillet varies typically from 16 . Fish flesh is dominated by the abundance of white muscle in relatively short segments. fins and a streamline body. In this chapter. The lipid content. which can be up to 67%. giving it its characteristically flaky structure. CHILLED AND FROZEN RAW FISH Associate Prof.2 . Fish can be divided into fatty and lean fish. Fish fillets are a poor source of carbohydrates. and other forms of aquatic animal as part of their ‘fish’ definition. but typically present a low fat percent when compared to other meats. lean fish are those fish that store most of their fat in the liver. offering less than 0. typically 3 and 15% of total weight.1. The lipid fraction is the component showing the greatest variation. typically fluctuates between 0.1 Definitions Fish are classified as any of the cold-blooded aquatic vertebrates of the super class Pisces typically showing gills. migratory patterns. Fish are an important part of a healthy diet since they contain high quality protein. In addition.
Spoilage describes changes that render fish objectionable to consumers. and maintained at or below -2 °C (normally below -12 °C) during storage. firmness. Organoleptic refers to qualities such as appearance. The proportion of each component is fixed when the mixture is introduced. and flavour based on the maturity. hence. at -2 to -4 °C. transportation and sale. during storage. transportation and sale. Frozen fish is fish that has been cooled to. and maintained at or below 7 °C. DMA is dimethyl amine.FISH AND SEAFOOD Chilled fish is fish that has been cooled to. Controlled-atmosphere packaging (CAP) refers to packaging in an atmosphere where the composition of the gases is continuously controlled during storage. nitrogen (N2) and oxygen (O2). Shelf life is defined as the time of storage before microbial spoilage of a fish is evident. marbling. and texture of the fish. colour. Superchilled fish is fish that has been cooled to. Evisceration is the removal of the viscera from a fish. juiciness. frozen or preserved. spoilage microflora describes an association of microorganisms that. 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. Fresh fish is raw fish that has not been processed. Spoilage potential is a measure of the propensity of microorganisms to render fish objectionable to consumers through the production of offensive metabolic byproducts. colour. Raw fish refers to fish that has not been cooked but excludes fish treated with curing salts and/or subjected to fermentation. usually carbon dioxide (CO2). and maintained at temperatures just below the freezing point. but not below 3 °C during storage. 2 . transportation and sale. Quality refers to palatability and organoleptic characteristics such as tenderness. odour and texture. through their development on fish. but no further control is exercised during storage. renders that fish objectionable to consumers.
molecular-based methods have not yet been used to address this issue. and those that although not autochthonous to the aquatic environment. posing a concern for seafoodborne illnesses (2). It is crucial to mimic the environmental physicochemical parameters when isolating bacteria from fish. However. whenever possible. Flavobacterium. More accurate. several culture media containing sodium chloride and more than one incubation temperature should be used. The number and diversity of microbes associated with fish depend on the geographical location. Shewanella and Pseudomonas. Gram-positive organisms such as Bacillus. standard culture-dependent methods can only recover between 1 to 10% of total bacteria present in any given sample. It is difficult to estimate how many microorganisms are typically associated with fish. microorganisms colonise the skin. as with other vertebrates. healthy fish is considered sterile and should not present any bacteria or other microorganisms.CHILLED AND FROZEN RAW FISH Vacuum packaging (VP) refers to packaging systems in which the air is evacuated and the package sealed. although these are influenced by water quality and feed. Gastrointestinal tracts and gills typically yield high bacteria numbers. Lactobacillus and coryneforms can also be found in varying proportions (1). and typical of the fish flora.2 Initial Microflora The subsurface flesh of live. potential human pathogens may be present in both scenarios. These pathogens can be divided into two groups: organisms naturally present on fish (Table 1. gills and the gastrointestinal tract of fish. In fact. the natural fish microflora tends to reflect the microbial communities of the surrounding waters. 1. Moraxella.I). the season and the method of harvest. In general. some species (most Vibrios) require sodium chloride for growth. are present there as result of contamination (anthropomorphic origin or other) or are introduced to the fish during harvest. For example. Human pathogenic bacteria can be part of the initial microflora of fish. On the contrary. Fish harvested from clean and cold waters will present lower bacterial numbers than fish from eutrophic and/or warm waters. The autochthonous bacterial flora of fish is dominated by Gram-negative genera including: Acinetobacter. since they heavily depend on the type of sample analysed and the protocol used for isolation. Clostridium. processing or storage (Table 1. It must be noted that mesophilic bacteria can rapidly overgrow psychrotropic organisms.II) (3). Members of the families Vibrionaceae (Vibrio and Photobacterium) and the Aeromonadaceae (Aeromonas spp. 3 . Micrococcus.) are also common aquatic bacteria.
vulnificus) 5 . Plesiomonas shigelloides 3 . 105 . 4).102 101 . estuarine environments.19 µg/kg bodyweight from < 102 . birds. numbers of pathogenic bacteria in raw fish tend to be low.>106 Salmonella spp. 4 .109 cells/g) * Some sources considered L. Vibrio cholerae Vibrio parahaemolyticus Vibrio vulnificus other vibrios Aeromonas spp.14 .37 °C High for most species. sewage. and risk associated with the consumption of seafood is low (2.FISH AND SEAFOOD TABLE 1.103 High (107 .I Indigenous bacterial pathogens typically present in fish (3) Organism Temperature range Estimated Minimum Infective Dose 00. human origin Minimum Infective Dose Variable depending on the strain (>102 cells/g) Staphylococcus aureus 105 .35 °C 8 .II Non-indigenous bacterial pathogens frequently present in fish (3) Organism Listeria monocytogenes* Primary habitat Soil.106 cells/g (exception: V.106 cells/g. stream water. In addition. during storage indigenous spoilage bacteria tend to outgrow potential pathogenic bacteria. Intestinal track of terrestrial vertebrates Human origin Fecal contamination Ubiquitous in environment Shigella spp. 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.1 µg toxin lethal dose Clostridium botulinum non-proteolytic type E Pathogenic Vibrio spp.37 °C Unknown Unknown TABLE 1.1 . However.26 °C 10 . Escherichia coli Yersinia enterocolitica 101 .0. and mud Ubiquitous. Toxin levels 0.
mackerel or herring. in which the health of each animal can be assessed prior to slaughter. offering better control of fish health prior to. or traps. chilled brine or refrigerated seawater at -2 °C. and spoilage. Fish may be eviscerated prior to storage at sea . These contacts will not only increase the bacterial cross-contamination between fish batches but will introduce microorganisms from other sources such as humans. and during harvest. and the killing process is designed to minimise stress. decks. The length of time that set nets have been in the water or the time trawlers’ nets are towed. boxes and/or baskets. If evisceration is carried out at sea. handling and processing Wild finfish are usually caught by net. ropes. The action of intestinal enzymes and activity of the gut bacteria on the flesh around the belly cavity may produce discolouration. with fish such as tuna and cod being more commonly eviscerated than sardines. with very little control over the condition of the fish at the time of death or the duration of the killing process. This contrasts greatly with the meat industry. Of particular concern is the use of wooden or soiled plastic containers for storage and unloading at the quayside. with the production of off-odours.CHILLED AND FROZEN RAW FISH Shelf life depends on the initial microflora on the fish. human hands and clothing. Physical damage such as loss of scales. Increasingly. freezing facilities (-18 °C) may be used to prevent the catch from deteriorating. in recent decades. The decision to eviscerate the catch at sea will depend greatly on the size of the fish and the duration of storage at sea.a practice that may have both advantages and disadvantages. potential contaminants added during handling and processing. care should be taken in removing all the gut contents and washing the carcass thoroughly prior to refrigerating.1 Processing and its Effects on the Microflora Capture. hook and line. and conditions of storage. In eviscerated fish. the cuttings provide areas of exposed flesh that are open to microbial attack. the fish may be stored in the vessel for periods ranging from just a few hours to several weeks in melting ice. however. icing or freezing. Used refrigerated brines can be contaminated with high numbers of psychrotrophic spoilage bacteria. and especially when fish is stored on board for longer periods. has an effect on the amount of stress and physical damage that the fish will suffer during capture.3.3 1. After capture. bruising and bursting of the gut will increase the number of sites open for bacterial attack and spread. In addition. finfish will almost invariably come into contact with nets. digestion and off-flavours in uneviscerated fish. birds and soil. aquaculture practices have been expanding worldwide. in which the bacterial load 5 . However. cortisone levels increase during prolonged stress and can alter the fillet quality. Inadequate circulation of chilled brines may result in localised anaerobic growth of some microorganisms. 1. and their re-use will increase the crosscontamination of other fish with such microorganisms. During capture and storage.
As with all foods. partial fermentation of sugars occur leading to lower pH. MAP extends the shelf life of most fishery products by inhibiting bacterial growth and autoxidation. they are usually process contaminants. Bacterial composition under MAP shifts from mostly Gram-negative to predominantly Gram-positive (lactic) bacteria. fish have a higher post mortem pH. Monitoring of the seawater for algal growth in order to limit the risk of algal toxin ingestion. allowing more growth of spoilage organisms. cleaning of the work environment. and of the quality of the water used for ice and to wash fish. will result in fully anoxic conditions. The atmospheric conditions surrounding a product may be passively or actively altered. Low oxygen levels have been shown to substantially prolong the freshness and quality life of refrigerated seafood products. use of effective detergents and disinfectants. and to limit the loss of quality (5). and specific spoilage organisms may use other terminal electron acceptors naturally present in the fish (trimethylamine-N-oxide (TMAO).FISH AND SEAFOOD can be substantial. the natural atmosphere is replaced with a controlled gas mixture (carbon dioxide. 1. from water activity (aw) through to physical damage. oxygen etc. Both carbon dioxide and low pH inhibit the growth of the typical spoilage bacteria such as Pseudomonas and Shewanella. by actively altering the composition of the 6 . 7).). Anaerobic atmospheres have less effect on fresh fish shelf life. but isolate the food in an environment different from the bulk atmosphere. potential spoilage bacteria are among the psychrophilic and psychrotrophic flora present on temperate-water fish before death (9). 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. Brochothrix thermosphacta and psychrotrophic lactic acid bacteria (LAB) can produce spoilage characteristics. Carbon dioxide is the most important gas in MAP of fish because of its bacteriostatic and fungistatic properties. ferric ion (Fe3+)). Physical barriers not only protect from physical damage. nitrogen. often in the absence of adequate refrigeration.2 Modified-atmosphere packaging A natural atmosphere rich in oxygen (21%) is responsible for oxidative processes and for all aerobic respiratory life. What is more. In MAP. for example. Packaging changes the intrinsic and extrinsic parameters affecting a product. By vacuum packing foods. in time. but if applied properly should extend the life of the product. These containers are also used for displaying the fish during auction at the quayside. careful and sanitary handling during processing is required to reduce the risk of contamination with potential human pathogens. However.3. a reduction in the oxygen tension is achieved which. not part of the normal flora of the meat animals (8). however. and minimising handling will all reduce microbial cross-contamination. In the absence of oxygen. if there is some oxygen demand from the product. These changes can be deleterious.
Growth and toxin production have been detected in artificially contaminated packs of whole trout after 1 week’s incubation at 10 °C (13). an inert and odourless gas. but can dissolve into fish liquids and deform packages. and the MAP of fish has more challenges to overcome as a result of: a comparatively large initial load of bacteria present. but only in recent years has it seen a marked expansion in use and market share. Replacement of oxygen with nitrogen. Gases used in MAP of fish most commonly include carbon dioxide and nitrogen. The principal effect of raised carbon dioxide-MAP is an extension of the 'lag' phase of the growth of the bacteria on the fish. without overt signs of spoilage. The microflora of meat is not the same as that of whole. MAP has been applied to fresh meat and fish with a resulting commercially viable extension in shelf life (10). the pathogens that may be able to grow before spoilage occurs. the higher pH and reduction potential (Eh) of the fish muscle. Of particular concern are the psychrotrophic type E and non-proteolytic type B and F strains. The (International) Codex Committee has published a Code of Practice for Fish and Fishery Products (CAC/RCP 52-2003). as they are able to grow at temperatures as low as 3. Many additives have been tried in conjunction with changed atmosphere. although less information on their effectiveness is available. This has been driven partly by increased consumer demand for fresh and chilled convenience foods containing fewer chemical preservatives. High concentrations of carbon dioxide have the most pronounced microbial effect. phosphates. and the promotion of a predominantly Gram-positive. a modified-atmosphere may contain any gas necessary for the desired effect. Sulphur dioxide. discolour pigmented fish (11). VP is one of the oldest forms of altering the interior gaseous environment of a pack. In the United States. but residual oxygen and other electron acceptors may be sufficient to allow oxidative spoilage of fish (12). Flavobacterium. Modified-atmosphere preservation of fish was first reported in the 1930s. The products. which are able to grow rapidly at low temperature. may not be considered ‘fresh’ fish. does inhibit some aerobic bacteria and reduce the rate of oxidative rancidity. sorbates and chelating agents such as ethylenediaminetetraacetic acid (EDTA) (11). after additive application. Micrococcus and Moraxella). the inhibition of common 'spoilage' bacteria (Pseudomonas. slower-growing flora (11). which provides guidance relating to vacuum or modified-atmosphere packaging for specific fish products (14). nitrous oxide and carbon monoxide have also been suggested as possible replacement gases in trace amounts for MAP/CAP. the National Advisory Committee on Microbiological Criteria for Foods (NACMCF) has published a number of recommendations on 7 . and the problems of muscle structure damage by the modified-atmosphere (11). including salt. gutted or filleted fish. The single most important concern with respect to the use of MAP is the potential for outgrowth and toxin production by C. botulinum. and increase in-pack drip (12).3 °C and produce toxins.CHILLED AND FROZEN RAW FISH surrounding gas.
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. greater advantages are seen. 3. trimethylamine and sulfides). For fatty fish. 2.g. from where drip loss and gaping of the muscle meant that organoleptically classified rejection occurred sooner than with no carbon dioxide addition (12).g. this problem occurs to a lesser extent with meat. 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). owing to the dissolution of cellular structures. The only processing dip to be used is 5% potassium sorbate and 10% sodium tripolyphosphate. TMAO) that can be reduced to produce 8 .FISH AND SEAFOOD the safety of MAP and VP refrigerated raw fishery products (15). may result in unacceptable colour and textural change and hence little shelf life extension (11). as already stated. oxidative rancidity is a significant source of spoilage and hence the complete removal of oxygen and the use of oxygen-impermeable films are preferred. Freshwater fish have different intrinsic bacterial flora and also do not contain such high concentrations of amines (e. but is recognised as a drawback of carbon dioxide application. however. leading to the recommendation that the sale of MAP/VP raw fishery products be allowed only when certain conditions are met. adequate shelf life and cooking requirements. In the United Kingdom. should be used. 4. It was also noted that organoleptic spoilage and rejection by the consumer should occur before the possibility of toxin production. The replacement gas should contain at least 40% carbon dioxide. The problem of water loss in MAP fish is common. however. adequate product labelling in respect of storage temperature. Subsequent storage must be at or below 3 °C.3 °C at all points from packaging onwards. botulinum. at all times. The addition of high levels of carbon dioxide to these fish. Only the highest-quality fish. and the use of a HACCP plan. 5. from both a microbiological and a chemical standpoint. The 1985 British Guidelines on MAP/CAP fish from the National Fisheries Institute state that: 1. Sensitive and accurate time-temperature indicators must be used to ensure that the product has not been temperature-abused. 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. For other less fatty marine fish. rejection due to alteration in fish texture with greater carbon dioxide makes this unsuitable (13). Although all fish species need to be treated separately. The growth of these bacteria is reduced by high carbon dioxide concentrations but. the Sea Fish Industry Authority has published guidelines for the handling of fish packaged in a controlled-atmosphere (16). These conditions include storage of the product at ≤ 3. use of high-quality raw fish.
texture. TABLE 1. and this pattern is the basis of freshness grading schemes. The eyes turn from convex and clear to concave and opaque.CHILLED AND FROZEN RAW FISH off-odours. these are summarised in Table 1. Loss of freshness and spoilage cannot be separated as processes. the skin turns from iridescent to dull and bleached with bacterial slime. and the flesh turns from bright and elastic to dull and soft (1).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. A.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).60% carbon dioxide with a balance of nitrogen for fatty fish have been recommended (16. smell and 9 . Various methods for whole fish freshness evaluation by a trained panel have been proposed. Gas mixtures of 40% CO2/30% N2/30% O2 for white fish and 40 . reactions caused by activities of the fish’s own enzymes. C (C=reject) Significantly. Spoilage of fish starts upon death due to autoxidation (oxidation of unsaturated lipids).4 1. 1. Over time. Degradation of whole fresh fish stored in ice generally follows a set pattern. 18) and are probably the most widely used. to brown and slimy with an intense off-odour. and metabolic activities of microorganisms present in the fish. B. rejection of whole fresh fish for human consumption may be made without chemical or microbiological evaluation. so lower carbon dioxide concentrations may have better preservative effects. Taste panel evaluation of cooked fish may be used to score or grade colour. as traditional spoilage patterns of the external organs are typically very distinct. the gills from pink and shiny. 17. evaluation of taste is unavoidable as the visual pattern of degradation of the eyes and gills is unavailable for analysis. but it is a commonly held view that loss of freshness is related to autolytic degradation and spoilage is more microbial in origin (1). or for that matter evaluation of the taste of the fish. with no smell.4. When working with fillets. loss of the fresh characteristics may be simply measured by comparative visual and smell analysis.
is significant and may even be responsible for the ultimate sensory rejection of fresh cod and other fish with high initial TMAO content (20. ADP. or simply a waste product present due to bioaccumulation (22).g. ammoniacal. fruity and/or bitter off-flavours. reduction in ATP and increase 10 . the cooked analysis of fish passes through four phases of spoilage: i) Delicate sweet. sea-weedy taste. The processes of degradation being analysed by the quality scoring methods above are a complex mix of physical. or its remaining shelf life (1). the muscles harden as residual adenosine triphosphate (ATP) is reduced and the myosin and actin filaments bind to form actomyosin (18). chemical. However. RT Freshmeter or Fishtester). all marine fish use/have TMAO (21). IMP. rigor mortis is the first noticeable change in the fish. the conductance generally increases. texture becomes soft and watery or hard and dry. rigor resolves. AMP. the Eh of fish muscle remains relatively high (20). %). the pH drop of the muscle is correspondingly less. bacterial reduction of TMAO to TMA. cabbage-like. the muscles relax again and the fish returns to a flaccid state. an intense odour compound. post-rigor values are typically in the range of pH 5. development of sickly sweet. HxR and Hx) gives a quantity (K-value. As the fish degrades. 23). iv) Enhancement of the spoilage characteristic of phase iii. the Torrymeter. After death. thus. iii) Traces of sour. Generally. a ratio of concentration of the hypoxanthine (Hx) and inosine (HxR) to total concentration (ATP. which has been ascribed a number of possible functions as: a trimethylamine (TMA) detoxified waste product. which increases from 0% towards 100% with time. as little endogenous reduction of TMAO occurs (18). From being flaccid. possibly slightly metallic. 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. 19). ii) Neutral taste. little flavour.5 (18. spoiled and putrid. 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. After death. During the initial storage. sulphurous and/or rancid smells. Because fish tend to have relatively little residual glycogen compared with mammals. this will generally correlate with the length and severity of rigor.8 to 6. an anti-freeze.FISH AND SEAFOOD taste of cooked fish. 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. These processes are strongly influenced by the physical conditions of storage. The process of ATP breakdown is used as an indicator of fish freshness (24). biochemical and microbiological actions. TMAO permits a high Eh to remain in the muscle tissue. To a varying extent. After some time. By measuring the concentrations of the six components.
so the measurement is effective only during the initial loss of freshness.CHILLED AND FROZEN RAW FISH in hypoxanthine by endogenous enzymes allow this measurement to be used for determining freshness (19). The Conway method (and variations of it) uses a strong inorganic base to volatilise the bases in the fish sample. The variation in post mortem fish pH may likewise influence the amount of bases being liberated to the air and consequently affect the odour characteristics of the fish. During spoilage. and a segregated weak acid to absorb them. does not show correlation between muscle pH and the amount of volatile bases contained within the fish at rejection. not to describe freshness. However. For some fish species. however. This measure is highly dependent upon the temperature of storage and may not reflect the rate of loss of quality equally at different temperatures. 26). Lipid oxidation and other oxidative changes lead to oxidative rancidity. these measurements are usually used to quantify the amount of spoilage. but rejection of whole fresh fish by sensory methods such as the EC grading scheme (32) is based upon non-specific odour detection and physical appearance. Lipid oxidation can be a result of enzymic action or a cascade reaction initiated by free radicals (26. the importance of microbial action increases with time (5. is later significantly reduced by bacterial action. 11 . 31). and are especially important in the spoilage of frozen fish. which may also be formed by bacteria (25). The major spoilage odours and flavours of fresh fish are undoubtedly principally microbial in origin. colour changes. Chemical degradation continues after the initial post mortem phase. therefore. 27) produced by aerobic respiration and other forms of metal ion reduction (28). for example the TMA fraction. the residual acid is then titrated. Oxidative rancidity is known to reduce the quality of fatty fish in particular (19. Evaluation of Total Volatile Base-Nitrogen (TVB-N. also termed TVN). Comparing results for different fish species. 18). a correlation can be made between the spoilage of the fresh fish and the production of TVB-N. however. as microbial spoilage is limited by the low storage temperature (1. as significant proteolysis is observed only during the latest stages of spoilage and after rejection (30. 7). Neither does the change in pH during storage correlate well with the production of TVB-N. the majority of volatile bases are produced from the soluble non-protein nitrogen of the fish (free amino acids and other low-molecular-weight nitrogenous compounds). or a specific fraction of the volatile bases. Quality indices based upon the products of microbial metabolism do not explain changes in quality until microbial growth produces measurable changes in the fish. using Conway diffusion chambers (29) allows determination of changes of mg-N/100 g fish. Volatile bases are the best-characterised chemical indicators of fresh fish spoilage. hypoxanthine.
by altering the gas atmosphere (extrinsic).5. For fresh fish. realisation that the bulk of the microbial population on newly caught fish does not cause off-flavours and off-odours stems from work 12 . 34). including the ability of the endogenous bacteria to grow at low temperatures and lower endogenous enzyme activity.1 Factors Affecting Fresh Fish Spoilage Temperature By far the most effective method of reducing the rate of whole fresh fish spoilage is temperature control (1. biochemical and microbiological reactions taking place within and on the fish. careful temperature control during storage is not only important in terms of quality loss. All chemical reaction rate kinetics (and thereby microbial growth) are temperature-dependent. The application of ice storage increases the shelf life of fresh fish from a matter of hours at ambient temperatures to days or weeks.5. The factors are interactive and cannot be completely isolated.2 Fish-spoilage bacteria The parameters affecting the multiplication of microorganisms in foods have been categorised into two general groups: intrinsic (inherent qualities of the food) and extrinsic (qualities of the food environment) (35). The sum of these changes may result in a different spoilage profile.FISH AND SEAFOOD 1.5. but also crucial for the assurance of consumer safety. the results of altering one parameter may have far-reaching consequences on others. Also. This increase has been reported to be moderately dependent on the temperature of the sea from which the fish are taken (33. 1. 1. Fish spoil as a result of the chemical. The concept of specific spoilage organisms (SSOs) is not new. 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 reasons for the differences between tropical. For fresh fish iced immediately after capture and continually stored in ice. the lower the temperature of storage. Not all the bacteria growing on a food will lead to the production of objectionable characteristics. 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 slower the spoilage processes proceed (within limits).and temperate-water fish spoilage rates may be many.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. a minority are often associated with the majority of the spoilage.5 1. For example.
Bacteria identified as being associated with the spoilage process of fresh fish are as follows: 1. During the 1970s. It is thus an important spoilage organism of gadoid fish such as cod. frozen) fish products.CHILLED AND FROZEN RAW FISH started in the 1940s. 1. 13 . more especially. It is also present in fresh water. 36). (9) and they are considered SSO of iced freshwater fish (35). its ability to reduce TMAO to TMA. work with inoculated sterile fish blocks resulted in identification of a specific minority of microorganisms.5.2 Shewanella putrefaciens Sh. mediated spoilage is characterised by ‘fruity’.3.10% of the total flora of fresh fish from temperate marine waters. i. These organisms are described as potential spoilers. obligate respiratory bacteria. the spoilage of a product may be strongly influenced by the conditions under which the product is held. esters and nonhydrogen sulphide sulphur-containing compounds such as methyl sulphide (36.1 Pseudomonas spp. The spoilage of freshwater fish is generally ascribed to the growth of Pseudomonas spp. although only since the late 1960s has it been realised that specific metabolites of Sh.3. aldehydes. The spoilage compounds associated with the growth of psychrotrophic Pseudomonas spp. Members of the genus are able to produce pigments. Pseudomonas spp. 35). The importance of Sh. and may play some role in the spoilage of freshwater fish. produce hydrogen sulphide (H2S) from cysteine. They are generally characterised as Gram-negative rods. form methylmercaptane (CH3SH) and dimethylsuphide ((CH3)2S) from methionine and produce hypoxanthine (Hx) from inosine monophosphate (IMP) or inosine. it is often isolated as about 1 .5.e. processed (e. putrefaciens to the spoilage of fresh marine fish has been recognised since the 1940s. The Pseudomonadaceae family represent a large and poorly defined group of microorganisms. 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 characterisation of spoilage of each product must be made before the identification of the responsible agent(s) can proceed. or faster than the rest of the flora of ice-stored fresh marine fish (9). putrefaciens spoilage of fish is due to its biochemical action on muscle. catalase-positive. putrefaciens growth may be used as indicators of spoilage (30). 37). on fish are diverse and in many cases species-specific.g. plus other characteristic compounds of the species responsible for spoilage. motile with polar flagella. It is able to grow as fast as. putrefaciens is considered an SSO of temperate-water marine fish species stored in ice. ‘oniony’ and ‘faecal’ odours from the production of ketones. which produced the characteristic spoilage compounds of the fish (30. As previously noted. oxidase-positive. and proteolytic and lipolytic enzymes that may affect the quality of fresh and. Sh. therefore. 35.
FISH AND SEAFOOD for which the most compelling evidence showing spoilage as a result of Sh. phosphoreum increased in notoriety when it was proposed that reduction of TMAO to TMA limited the shelf life of MAP cod fillets (12). mesophilic strains of Sh. Shewanella denitrificans. This wide-ranging respiratory capability is thought to be unique (39). putrefaciens lies in its chemotaxis: the bacterium. It is also able to produce hydrogen sulphide and a range of other off-odour compounds. a population of Ph. it was proposed that this organism. with several new species being described. putrefaciens may grow in the absence of oxygen using alternative terminal electron acceptors (ATECs). The localisation of the reductase on the outer cellular membrane offers the bacteria a solution to this problem (40). Another distinct feature of the physiology of Sh. it is strictly respiratory. unlike many other motile bacteria. Initially. was capable of being in a significant numerical minority on the MAP fish but still able to yield the majority of the TMA thought to be responsible for the rejection (23. Shewanella olleyana. and Shewanella profunda were described. Sh. Shewanella gelidimarina. like all members of the Pseudomonadaceae. does not show carbon-source chemotaxis. Ph. phosphoreum sufficient to cause spoilage was found. Sh. owing to its cell size and activity. The prevalence of the bacterium in so many environments. 1. putrefaciens has been studied. The ferric reductase activity of Sh.3. In fact. Shewanella affinis. at the time of rejection. This type of intoxication occurs when bacteria convert the histidine 14 . the bacteria have a significant problem to overcome . Because no other TMAO-reducing bacteria were present in sufficient numbers to produce the quantities of TMA that were related to rejection. but Sh. Shewanella frigidimarina. putrefaciens were required. Shewanella waksmanii. Shewanella oneidensis. Research has shown that approximately 107 cfu/g of Ph. phosphoreum has also been identified as responsible for histamine fish poisoning. although. as the ferric iron is insoluble. 38). additional psychrotrophic species such as Shewanalla baltica.how to reduce a molecule that they are not able to take into their cells efficiently. baltica has been identified as the main H2S producer in cod during cold storing (37). In terms of its taxonomy. 41). and Shewanella aquimarina. putrefaciens were identified as Shewanella algae. Later. and its metal ion reduction ability has led to intensive investigation into its role in iron and sulphur cycles. whereas >108 cfu/g Sh.5. but does show strong chemotaxis up gradients of most of their alternative electron acceptors (39). Ph. Shewanella livingstonensis.3 Photobacterium phosphoreum Recognised for some time as being present on spoiling fish (30). phosphoreum were required for 50% of taste panellists to reject a sample. putrefaciens growth has been collected (12. and recent taxonomic description using modern molecular methods bifurcated this species. Sh. they are heterogeneous. In 50% N2/50% CO2 MAP cod. putrefaciens strains are characterised as microaerophiles or anaerobes. 41). putrefaciens was not present in such numbers (23.
human viruses. Recent studies have investigated the dominance of B. B. B. where a particular 15 . especially type E botulinum intoxication.1 Clostridium botulinum C. but with proper cooking. 6). bacteria and higher parasites all present some risk associated with fish consumption (1. botulinum recorded up to 1963. 1.3 °C (Group II) and these are non-proteolytic (i. Review of outbreak data suggests fresh fish to be safe. it is bacterial risks that increase after capture of the fish. 1. Adult disease is caused by the production of the botulinum toxin. There are few human bacterial pathogens that can cause primary infections or disease and are capable of persisting in the aquatic environment. thermosphacta did not produce off-flavours until 2 . D. a neurotoxin that causes flaccid paralysis and is associated with a variable (44) mortality depending upon dose.6. 5). 1. The heterogeneity of the types (A. presents a potential risk. C2. thermosphacta on spoiling fish in a 40% CO2/30% N2/30% O2 MAP (42).3 days after having reached 108 cfu/g (42). (and very rarely F and G) have been reported to cause human disease. When inoculated into VP corned beef and sliced ham. thermosphacta in the spoilage of some MAP fish.CHILLED AND FROZEN RAW FISH present in fish into histamine. 75% (with a 35% mortality rate) occurred in Japan. 5. E. MAP studies have also demonstrated this organism’s sensitivity to oxygen and have shown that they are also inhibited by high CO2 concentration (43). However. The remaining few bacteria present a major risk involved with the consumption of raw seafood such as sushi or oysters. thermosphacta is a well-characterised psychrophilic spoilage organism of meat.3. Of these. E and F include bacteria with minimum growth temperatures of approximately 3. C1.5. Fewer still are capable of growing on fresh fish. Huss (1.e. they do not produce significant product spoilage). these risks are substantially reduced (1. F and G) is well documented (44). E. B. phosphoreum strains have great capacity as histamine producers even under refrigeration conditions. a convenient but diverse species of bacterium with a number of different types. previous exposure and access to supportive treatment.6 Pathogens: Growth and Survival Endogenous chemicals. botulinum. Some Ph. this is probably due to spoilage occurring before toxin elaboration. 5) reports that fresh fish consumption has never been shown to cause human botulism. lightly preserved fish products are associated with botulism. Growing evidence suggests a role for B. types B. Of 404 intoxication outbreaks of type E C. including antisera.4 Brochothrix thermosphacta and lactic acid bacteria B. Types A. algal toxins. age. Acetate production has been reported as a good indicator of spoilage by this organism.
Japan. salted or pickled) fish have also been associated with botulism. there is no specific guideline that describes a minimum level of V. again. and North America. 1978 . A unique clone of V. Human disease caused by the organism has only been observed in conjunction with marine bivalve (mostly oysters) and some crustacean consumption.3 °C) will eliminate all risk. the risk posed to the consumer of fresh fish. parahaemolyticus is a marine organism. Proper chilling. In Japan. a Japanese boiled and semi-dried sardine dish. parahaemolyticus O3:K6 is responsible for many of the recent V. France. the majority are associated with products not cooked immediately before consumption. were associated with finfish between 1978 and 1998 (46).g. 1. which was probably contaminated from an uncooked food source or an excreting foodhandler. parahaemolyticus infections since 1996 and is referred to as the pandemic strain (47). The risk of toxin formation before apparent spoilage has been studied for some MAP fish (45). It is generally undetectable in marine water below 19 °C but may grow in culture at temperatures as low as 5 °C and on food at 10 °C.g. 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. Other lightly or semi-preserved (e. parahaemolyticus outbreaks. storage and preparation (11). 5). but as fresh fish are often cooked before consumption and the botulinum toxin is heat-labile. which is believed to be required for virulence. V. USA. Currently. Southeast Asia. Russia. use of post harvest treatments and/or cooking of fresh seafood will reduce risks. V. parahaemolyticus in sea water fish and shellfish that could potentially be hazardous to humans. accounted for the majority of cases (1. including epidemics in India. Proper temperature control (<3. the majority of outbreaks are caused by consumption of raw fish products (e. only shellfish are associated with the disease and no reported cases by the Center for Diseases Control. Only about 1% of marine isolates produce a thermostable haemolysin. there remains only an ‘extremely small’ risk of intoxication if reasonable precautions are taken during handling. Despite the ubiquitous nature of type E C. The first report of the organism being a foodborne agent was with shirasu. whether stored aerobically or in modified-atmospheres. vulnificus is associated with warm marine and estuarine waters.FISH AND SEAFOOD fermented raw fish product that is consumed uncooked.2 Vibrio parahaemolyticus and other vibrios V. This strain has been responsible for 50 to 80% of all V. and proper cooking (boiling for 1 minute. smoked. or cooking at 80 °C for 5 minutes) (44) will substantially reduce risk. which can cause human gastroenteritis. Generally. especially in individuals with underlying diseases (patients suffer from immune and liver diseases or blood 16 . is small. 11).6. parahaemolyticus cases have increased across the world. due to the cultural preference for raw fish dishes. I-sushi. Since 1996.2005). and when in contact with contaminated water (no CDC reported cases from fish. The bacterium causes primary septicaemia. sushi) (5).
Similar problems as those encountered for Aeromonas spp. are present on fish and can grow to significant numbers during storage (1. 5. In the US.6. L. the causative agent of cholera. and proper cooking will substantially reduce risk. Some isolates are true psychrophiles.000 17 . but the bacterium is known to survive and grow in the shallow marine. although lack of data concerning infectious doses and the possibility that foodborne virulence is linked to the immune state of the host mean that full risk assessment is impossible.20 °C. Most cases have been sporadic rather than associated to large outbreaks. in assessing the risk of fishborne infection by P. which is often fatal (>50%). with minimum growth temperatures around 0 °C and optima of 15 .4 Listeria monocytogenes L.3 Aeromonas Aeromonas spp.6. monocytogenes incidence has been between 0. It is known that Aeromonas spp. infection. 1. shigelloides are encountered. others are psychrotrophic mesophiles (49). It is particularly associated with disease following consumption of raw oysters from warm sewagepolluted waters. Despite this. is historically associated with faecally contaminated water. however. are associated with human diarrhoeal illnesses.. Although the psychrophiles are unable to grow at body temperature. The major virulence factor appears to be the production of toxins with enterotoxic. but proper cooking should substantially reduce risk of infection. coli on isolation media (50). There is no risk associated with the consumption of properly chilled and cooked fresh fish. possibly due to under-reporting because of their likeness to E. sodium channel blocking and haemolytic activity (48). and fish and shellfish are probably the major reservoirs for the organism (50). shigelloides is similar to Aeromonas spp. though it is a true mesophile with a minimum growth temperature of 8 °C and shows consequential seasonal variation in its environmental isolation. their potential to produce disease-causing exotoxin in food has not been fully examined (48). 1. and especially Aeromonas hydrophila. 17). The bacterium has been documented as being responsible for a few fishborne outbreaks of gastroenteritis. V. listeriosis cases have been reportable to the CDC. no more than circumstantial evidence exists linking the consumption of seafood to Aeromonas spp. in its habitat.55 cases per 100. cytotoxic.26 to 0. They are aquatic organisms with an epidemiology that is yet to be fully understood. cholerae O1. there is no risk from the consumption of properly handled and cooked fresh fish. P. and especially estuarine environment. Chill storage may not eliminate growth. Again.CHILLED AND FROZEN RAW FISH disorders). monocytogenes has been well documented as a foodborne human pathogen. Since 2000. proper chill storage will eliminate any risk associated with this organism.
6.pregnant women. neonates.FISH AND SEAFOOD persons.6 Parasites Helminthic parasites can occur extensively in finfish but very few are capable of infecting humans. it is not apparent to the consumer and it cannot be destroyed by cooking. AIDS patients and patients undergoing immunosuppressive therapy) and a lack of data covering infectious doses.2002 (55)) and is also common worldwide. 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. foetuses.6. there is a far greater risk of unacceptable histamine levels being attained before sensory rejection. but cooking will significantly reduce the risk of infection.. some Vibrio spp. full risk analysis is not possible (52). 1. Owing to the dependence of the virulence of this organism on the immune state of the host (usually requiring lowered cellular immunity. Fortunately. The organism poses a serious risk in chilled products not cooked before consumption. there is a possibility for growth of the organism on chill-stored fish. leads to the production of histamine.48 minutes in salmon have been reported (53). within high histidine-containing fish (especially members of the Scombridae and Scomberesocidae families) is responsible for an intoxication termed scombroid poisoning (54). Klebsiella pneumoniae and Hafnia alvei. Photobacterium spp. alcoholics. Infections are mostly associated with the consumption of raw or 18 . Being a psychrotroph. 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. Low-temperature storage of potentially toxigenic (<5 °C) fish at all times is the most effective way to control histamine production (6). 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. at risk groups .5 Scombroid fish poisoning The production of histamine and other biogenic amines with human immunological activity. 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.. Decarboxylation of histidine by a wide range of bacteria including Enterobacteriacae. D60 of 1. It is regularly isolated from seafoods (51). Of fish-transmitted human diseases scombroid poisoning is commonly reported in the US (118 outbreaks with 463 cases reported by CDC 1998 . scombroid poisoning is usually a mild intoxication and it is not a significant cause of death.. especially pickled fish. but proper cooking will reduce any risk. Lightly preserved products.98 minutes in cod and D60 of 4. It is environmentally ubiquitous but its true frequency in the marine environment is not well studied. Like other toxins. If fish are not properly refrigerated. but more especially by Morganella morganii. Clostridium spp. and Lactobacillus spp. 1.
Cooking properly immediately before consumption is also an effective way of reducing or eliminating risk of fresh fishborne disease.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. risk of human disease caused by natural bacterial contamination (not through faecal pollution) of fresh fish is extremely low... TABLE1. A limited number of nematodes and trematodes found in finfish have also been identified as a cause of human disease (56). aureus 10 7 7 2 3 3 19 5 5 5 0 2 2 0 102 103 103 104 . which in many cases will eliminate the risk. 444. No. long storage under refrigeration or one of the former prior to freezing. including parasites and/or quantity of their toxins / metabolites per unit of mass. it may be desirable to test for Salmonella and V.H.IV) (57). Gram L. An increase of APC to levels in excess of 106 cfu/g is usually indicative of inadequate refrigeration.IV Sampling plans and recommended microbiological limits for fresh and frozen fish (Adapted from ICMSF 1986) Limit per gram or cm2 Test Aerobic plate count E. 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. 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. 2003.CHILLED AND FROZEN RAW FISH mildly processed fish such as sushi. coli counts where this method is preferred. volume. 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. parahaemolyticus (see Table 1. Ababouch L. EC.1997. parahaemolyticus Staph. area or lot (CAC. Rome. coli proposed by the International Commission on Microbiological Specifications for Foods (ICMSF). Testing methods should be practical and the enforcement of such criteria should translate as a reduction of potential microbiological risks to consumers. especially in warm-water areas and where fish are to be consumed raw. 1997) Huss H. Faecal coliform counts may be used instead of E. The most effective control of all bacterial risks is continual chill storage in ice (0 °C) or freezing. Criteria should set standards that are attainable by the currently accepted GMP and applied only when there is absolute need for it. 1. FAO.
. Codex Alimentarius Commission. References Huss H... Quality and quality changes in fresh fish. 87-118. in FAO Fisheries Technical Paper No. Huss H. Food Control. 13. 7. 107-27. 91-8. International Journal of Food Science and Technology. Incidence of foodborne pathogens in European fish. 10. 5. Food and Agriculture Organisation. 5 (1). Gram L.A. 9.. 11.H. 1995. Sivertsvik M. Spoilage and shelf life extension of fresh fish and shell-fish.. International Journal of Food Microbiology.. Huss H.. 1996. 121-37. in FAO Fisheries Technical Paper No. Feldhusen F.N..R.. Theoretical analysis of two hot water cabinet systems for decontamination of sides of beef. Huss H. Fletcher R. Code of Practice for Fish and Fishery Products (CAC/RCP 52-2003). Review: Assessing and managing risk due to consumption of seafood contaminated with microorganisms. 2000. 283-94. 1994. 348.P. 1989.J.K. Gram L. 6. Ed. Kirby R. Microbes and Infection. Smith J.FISH AND SEAFOOD 1. activities and safety. Critical Reviews in Food Science and Nutrition. International Journal of Food Microbiology. 243-60. Nychas G. 87121. 27. Spoilage and shelf life of cod fillets packed in vacuum or modified atmospheres. A review of modified atmosphere packaging of fish and fishery products – significance of microbial growth. 8. 1651-60. 2001. Davies A.K.8 1. Gibbs P. Ashie I. M. Jehanno D.R. Rome. 2. 1996. 8 (2). Assurance of seafood quality. Control of indigenous pathogenic bacterial in seafood.H. Food and Agriculture Organisation of the United Nations. Jeksrud W. 33. Huss H. Food Control. International Journal of Food Sciences and Techonology. 24. 334. 36. parasites.E. 5-7. Dalgaard P.J.J. 1997.. Journal of Food Safety.A.E. Codex Alimentarius Commission. Davies A. Armstrong D. 2003. Italy. Incidence and growth of psychrotrophic Clostridium botulinum in foods. 1994. 12. Shelf life extension and safety concerns about fresh fishery products packaged under modified atmospheres: a review.. 19..S. and natural toxins.H. 2002. 1993.. Simpson B. Ahmed F.. Rhodehamel E.H.. Food Control. 1992. 1992. 20 . 37. 67-71. 12.R.H. International Journal of Food Microbiology. Capell C.R. Rome.T. Food and Agriculture Organisation of the United Nations. 12.A. The role of seafood in bacterial foodborne diseases. Reddy N. Kautter D. Microbiological spoilage of fish and fish products. Ed. Rosnes J. Davey K. 3. Food and Agriculture Organisation. 4. 14. 2.. 291-304.
Surette M. 27. Gill T. Gill T.A.J. Changes in purine nucleotide of red lateral muscle of rainbow trout. 24. Hebard C. Huis in't Veld J.C.E.A.M. 1985. 1933.. 21.. 29...H. in Advances in Seafood Biochemistry. 121-37.J.. 1996. Autolytic changes in fish post mortem. 354-8. Washington D. 41-50. 36. Martin R. 1987.M. Byrne A. Biochemical Journal. International Journal of Food Microbiology. Davies A. 1-18. 1959.. 1992. Hebard C. Huss H. 22-36. International Journal of Food Microbiology. 1995. International Journal of Food Microbiology. Vacuum or Modified Atmosphere Packaging for Refrigerated Raw Fishery Products. 26.. Biochemical basis of post-mortem nucleotide catabolism in cod (Gadus morhua) and its relationship to spoilage. 1995. Qualitative and quantitative characterisation of spoilage bacteria from packed fish. Conway E.H. in Chemistry and Biology of Marine Food Products. 27.. Lipid oxidation in seafood. Ed. Food and Agricultural Organisation of the United Nations. Microbial and biochemical spoilage of foods: an overview. Shewan J. March 20. 19.. Reginstein J. 1995.E. http://seafood. Rome. Arai K. Yajima T.G. 1992. 1995.. Martin R. 305-17.CHILLED AND FROZEN RAW FISH 15. Quality and Quality Changes. Hendrie M. Part I . 28. 23.N. National Advisory Committee on Microbiological Criteria for Foods.. Shelf life extension of fresh fish . Flick G. 22.E.J. 245-67. Adopted.H..J.a review. Quality and Quality Changes. 2007. 26. 1971. Ratnayake W. 1. Saito T.. 419-29. Eds. Dalgaard P.. 184. in Fresh Fish.spoilage of fish.R. Journal of Agricultural and Food Chemistry. Ackman R. Pedrosa-Menabrito A.. Non-enzymatic oxidation of seafood lipids.htm Anon.M. Børresen T. Occurrence and significance of trimethylamine oxide and its derivatives in fish and shellfish.edu/haccp/compendium/chapt08. 21. Schwall D.. Leblanc P. An absorption apparatus for the micro-determination of certain volatile substances.. Guidelines for the Handling of Fish Packed in a Controlled Atmosphere. Letters in Applied Microbiology. Eds. Bacteria active in the spoilage of certain seafoods. Khayat A. Journal of Food Quality. Herbert R.ucdavis.J. 33. Journal of Applied Bacteriology. Microbiological spoilage of fish and fish products. Fate of Aeromonas and Yersinia on modified-atmospherepackaged (MAP) cod and trout.H. 1996. National Advisory Committee on Microbiological Criteria for Foods. 30. 1415. AVI Publishing. in Fresh Fish. 1988. 37. 25.M. Huss H.E. Ed. Flick G. Huss H. Rome.S. 42-54. Slade A. 34. 117-27. Sea Fish Industry Authority. Westport. 20. 130-2. Technomic. 1983. 21 16. Lancaster. Food Technology. The micro-determination of ammonia. 1982. Martin R. Edinburgh.. Gram L. 17.J.J. 18. Nature. 11. Chemical composition.E. Food and Agricultural Organisation of the United Nations.J. . Gibson D. 33. Flick G.E. 19-22.
262-6. 328-9. Role of proteolytic bacteria in fish spoilage. 2002. 39. 1995.. The inhibition by CO2 of the growth and metabolism of microorganisms. 1982. 311-43. Environmental Biotechnology.. 32. and regulation.FISH AND SEAFOOD 31. FAO Fisheries Report. Quality and Quality Changes. Rome. Saffarini D. Myers J.J. 62-70.S. International Journal of Food Microbiology.R. Dalgaard P. 3429-38. 4. Lee J. Kell D. COUNCIL REGULATION (EC) No 2406/96 of 26 November 1996 laying down common marketing standards for certain fishery products http://eurlex. 9. Annual Review of Microbiology. Vogel B.M. Huss H. 13.H. 36. 1994.. 72-80. 1984. 36.M. 1992. 317. Occurrence and importance of proteolytic bacteria as spoilage indicators. Jørgensen B.L. 41. Localisation of cytochromes to the outer membrane of anaerobically grown Shewanella putrefaciens. Cohen D..H. 1996. Libbey L. Scanlan R.H. 51-62.E... Volatile compounds produced in sterile fish muscle (Sebastes melanops) by Pseudomonas putrefaciens.do?uri=CONSLEG:1996R2406:20050602:E N:PDF Sumner J. 48... The effect of storage temperature on shelf life.A. 38. Archiv für Lebensmittelhygiene. 44. 71. Dalgaard P. 1973. a dominant microorganism in Mediterranean fresh fish (Sparus aurata) stored under modified atmosphere. Oxford. Nychas G. physiology. Myers C. 43. 33. Venkateswaran K.. 34.. in Fresh Fish. Blackwell Scientific Publications... Processes in Pathology and Microbiology. 2005.J. Sumner J..europa. II. 26. Gram L. Applied Microbiology. 18-21. Journal of Applied Bacteriology. Miller A. Identification of Shewanella baltica as the most important H2S-producing species during iced storage of Danish marine fish.. 6689-97. Brochothrix thermosphacta. Applied and Environmental Microbiology. 109-36. Fish spoilage bacteria – problems and solutions..F. 61-6. Dixon N. 22 .. Do Tropical Fish Keep Longer in Ice Than Temperate Fish: The Circumstantial and Definitive Approaches. Brady P. Nealson K. 37. Food and Agricultural Organisation of the United Nations. 174. Iron and manganese in anaerobic respiration: environmental significance. 33. Journal of Bacteriology. Growth and activity of Shewanella putrefaciens isolated from spoiling fish. 67. and an Achromobacter species. 323-9. Gorczyca E. Pseudomonas fluorescens. Taussig M. Do fish from tropical waters spoil less rapidly in ice than fish from temperate waters? Food Technology in Australia. 40. Magno-Orejana F. 334.R. 35.M. 42. Gram L. Rome. Satomi M. 1985.eu/LexUriServ/LexUriServ. Italian Journal of Food Science. Karnop G. Huss H.H. 1989. 1989. 1984. Ed. Drosinos E.B.
303-12.CHILLED AND FROZEN RAW FISH 45. Bishop R. Risk of growth and toxin production by Clostridium botulinum non-proteolytic types B.W. 17-34. 58.P. Ljungh A. Microorganisms in Foods 2. The International Commission on Microbiological Specifications for Foods. 57. Wilson S. 48. Golden D.. Huss H. International Journal of Food Microbiology. PCR detection of a newly emerged pandemic Vibrio parahaemolyticus O3:K6 pathogen in pure cultures and seeded waters from the Gulf of Mexico. 181-96.K. 49. Embarek B. 2000. Presence. Modern Food Microbiology.. Panicker G. Embarek B. Springer.L..P.N. 85-95. International Journal of Food Microbiology. Painter J. Loessner M.. Blackwell Scientific Publications. Smith J. Whiting R. Griffin P.... Journal of Infectious Diseases. Heat resistence of Listeria monocytogenes in vacuum packaged pasteurised fish fillets.M.H. Lynch M. 2000. 53.. Mac Rae I. 51 (4)... 1991. Kluwer Academic/Plenum Publishers. 51.. in Microorganisms in Foods 6. 55 (SS10). 100-3.A. Bean N. Surveillance for Foodborne-Disease Outbreaks . Microbial Ecology of Food Commodities. 12. The International Commission on Microbiological Specifications for Foods. 1986.K.J. 50. Experimental evidence for toxin production by Aeromonas hydrophila and Aeromonas sobria in a meat extract at low temperatures. 12. 2194200.. E and F in salmon fillets stored under modified atmospheres at low and abused temperatures. Applied and Environmental Microbiology. 1987. Aeromonas and Plesiomonas as food.. Ed. detection and growth of Listeria monocytogenes in seafoods: a review.. Myers M.United States..C.J..K. 1991. Ray B. Morbidity and Mortality Weekly Report. The International Commission on Microbiological Specifications for Foods. 1-37. 46.A. 2005.L. Woodruff R. Thompson S. Garcia G. 47.M. 2003. 2005. Bej A. Altekruse S.. New York. Miller A. 330-6. 181-8. MacKinnon L. Food Technology. 23. Use of risk assessment to reduce Listeriosis incidence.. 2006.C. Majeed K. International Journal of Food Microbiology. Hammond R. 1993. Vibrio parahaemolyticus Infections in the United States.and waterborne pathogens. International Journal of Food Microbiology. Fish and Fish Products (Microorganisms in Fish Products).H. Histamine fish poisoning revisited. 1998-2002. USA.. Sampling for Microbiological Analysis: Principles and Specific Application. Olley J.. 1994. Oxford. 54. International Journal of Food Microbiology. Slutsker L. 181. 55. 172-249. Wadström T. Journal of Food Protection. Lehane L. 52.M. Braden C.. 56. Daniels N.. 20. Genigeorgis C. 23 .. Lindroth S. 50.. 1661-6. 1973-1998. 69. Jay J. 1997. 1-34..
C. Ross T.. Sumner J. Jones Y. Microbiological quality and shelf life prediction of chilled fish. Smith G. Cann D.P. Kitts D. Eds. Akiba M. Huss H.. Journal of Food Protection. Borgstrom G. Houston N.. Lancaster. 1985. The Microbiology of Seawater Fish. Further Studies on Marine Fish Stored Under Modified Atmosphere Packaging. Ed. Studies on the microbiological ecology of mackerel stored by the method of partial freezing. Picard G.H. International Journal of Food Microbiology.H. Food and Agriculture Organisation of the United Nations. Ed. Taoukis P. 62-70. Dalgaard P. in Fish as Food. Food and Agriculture Organisation. Rome. Hobbs G.. Torry Research Station.9. Detection of specific spoilage bacteria from fish stored at low (0 °C) and high (20 °C) temperatures. 317 (Suppl. International Journal of Food Microbiology. 13-32.A.. Application of predictive microbiology to assure the quality and safety of fish and fish products. Olley J.. McMeekin T.H. Villemure G. Huss H.1 Further Reading Processing Anon. 1989.. Bulk storage of cod fillets and gutted cod (Gadus morhua) under carbon dioxide atmosphere.M. Journal of Food Science.. 35. International Journal of Food Microbiology.. 1986.G. Torry Research Station of the Ministry of Agriculture. 55. 73-89. Simard R. Fisheries and Food. Estimation of the bacteriological quality of fish by automated conductance measurements.E. Fisheries and Food. 1989. Gram L.. Florida. in Seafood Science and Technology.. International Journal of Food Microbiology.1983.)..2 Spoilage Shewan J. Cho Y.9. 1.M... 127-34. Gibson D. 1992. Trolle G. 1992. Aberdeen. Ogden I.S. Changes in microflora and chemical compounds in mackerel stored by partial freezing. Shahidi F. 1984. 1961.. 1.FISH AND SEAFOOD 1. 271-85.L. Mathematical Modelling Used for Evaluation and Prediction of Microbial Fish Spoilage..9 1. Gibson D. Torry Research Station of the Ministry of Agriculture. 51.. Academic Press. Ravn Jørgensen B.M.. 741-50. Bulletin of the Faculty of Fisheries (Hokkaido University). in Torry Advisory Note No 93. 6.D. Magno-Orejana F. Prediction of shelf life and safety of minimally processed CAP/MAP chilled foods: a review. 1984. Handling and processing of scad.. Do Tropical Fish Keep Longer in Ice than Temperate Fish: the Circumstantial and Definitive Approaches.. 65-72. 15. 1997. Labuza T. Shinano H.D. in FAO Fisheries Report No. 4. 1. 317-21. Aberdeen. Technomic Publishers. 295-307. 24 . 1988. Huss H.
. in Fish processing technology. Journal of Applied Bacteriology. in Quality Assurance in the Fish Industry. International Journal of Food Microbiology. London.. 72.H. Trolle G. Eds. Gram L. Jacobsen M. Klausen N. 322-6. 1992. Predicting the shelf life of cod (Gadus morhua) fillets stored in air and modified atmosphere at temperatures between -4 °C and +16 °C. Modified atmosphere packaging of fish and fish products. 25 . 20023. Huss H. 1987.K. pre-incubation at low temperature and pH on the thermal resistance of Aeromonas hydrophila. 59. 137-46. Garcia M.eu/LexUriServ/LexUriServ.J. Liston J.europa. Applied and Environmental Microbiology. A rapid method for detection of histamine producing bacteria.9.4 Standards and criteria Council Regulation (EC) Number 2406/96 of 26 November 1996 laying down common marketing standards for certain fishery products. Hall G. 5.9. Sala F.M. International Journal of Food Microbiology. 65-72.3 Pathogens Condon S. 1987. 1. Blackie Academic & Professional...H. Inhibitory effect against pathogenic and spoilage bacteria of Pseudomonas strains isolated from spoiled and fresh fish. Effect of culture age. Detection of specific spoilage bacteria from fish stored at low (0 °C) and high (20 °C) temperatures..do?uri=CONSLEG:1996R2406: 20050602:EN:PDF Davies A. 1992.H.. 1997. 1993. http://eur-lex.. Huss H. Elsevier Science.. 479-88.R. 2197-203. 4. Amsterdam. 1. Otero A.CHILLED AND FROZEN RAW FISH Einarsson H.L. Ed. Gram L. Huss H.
or could be disguised by sauces. although recent developments in improved supply chains. it does not necessarily apply to the toxins that may have been produced.g. The same used to be said for the difference between fresh and frozen products. CHILLED AND FROZEN PREPARED FISH PRODUCTS Dr. Few seafood products are sold ‘ready-to-eat’. and crustacea such as shrimp and crabs. the exceptions being Sushimi (high quality raw fish). products of lesser quality. Simon Derrick The Grimsby Institute Humber Seafood Institute Europarc Grimsby DN37 9TZ United Kingdom 2. This is no longer the case with many processed products now available in stores. 27 .g. 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. colourings etc. This has the result that any microbes present in the product have the capability to grow (with subsequent spoilage) unless strict temperature controls are applied.). Sushimi tuna). seafood (with the exception of some crustaceans) remains uncooked until the point of consumption. due to the early cessation of spoilage. Even though heating prior to consumption will eliminate pathogenic microbes. clarity of eyes. smoked (hot) mackerel.2. but still acceptable and meeting all safety criteria. 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. smoked (cold) salmon.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. emphasising the high quality of the seafood and the gourmet nature of the prepared products. smell etc. colour of gills. are used in chilled and frozen prepared meals since many of the attributes on which quality is assessed are not present or noticeable (e. The majority of seafood products produced require a cooking stage before consumption.
Listeria spp. The expected shelf life attainable for this format is usually up to 8 days. and are well known for their diet consisting of waste materials. shellfish and molluscs. they are usually supplied filleted with head on. 2. where the preparation has already been carried out. and this has produced a greater availability and diversity of products in the marketplace. They can be with or without skin and may contain pin-bones although the major skeletal bones are removed during filleting. but these are now finding favour as products in their own right. Seafood is the term used descriptively to include all categories of product. This includes all categories of seafood including trout and other freshwater species.2 Definitions The traditional focus of the chilled and frozen fish market has shifted over the last few years. The market spans fresh fish counter sales and frozen recipe dishes with fish as the predominant source of protein. the processing industry has developed a wide range of value added products. Doubts have been raised over the microbiological status and handling practices of reef-caught fish. etc. It is sold from wet fish counters on ice or pre-packed in modified atmosphere packaging (MAP). Frozen fillets may have a surface layer of 28 . Marketing-style descriptions and process definitions are presented below to acquaint the reader with the variety and forms of seafood available. 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.. Nevertheless. Warm-water/reef-caught fish were initially supplied to add colour and variety to wet fish displays. but should be equivalent to cold-water products. as they tend to lose colour and sheen after defrosting. in particular.. have shown the increasing importance of convenience foods.FISH AND SEAFOOD Recent trends in the market for food and seafood products. which can be farmed. rather than frozen. prawns. At the same time. and pathogenic Vibrio spp. or in MAP depends on catch conditions and treatment. There is also a limited market for filleted material. Microbiological hazards include Salmonella. They are typically flown into export markets. and the consumer only has to heat (cook) the product. often using the simplicity of a microwave oven. Wet or chilled/frozen whole fish are whole fresh fish that may or may not have been filleted. Prepared natural fillets are fillets of fish without any additional coating. including freshwater fish. molluscs. and these are increasingly being presented with an ever-expanding range of accompaniments. Cold-water fish microbiological specifications are inappropriate for assessing freshness of warm-water fish. Shelf life on ice. from both wild capture and aquaculture sources. they are usually of a better standard than species such as tilapia.
and the product then shaped and heat-processed to cook the product before being chilled. mackerel. which is approximately 30% (or 1 . These products are packaged in separate high-care facilities to minimise contamination with microbes either from the workforce or from raw products. Added colours can be included with the brine solution to meet specific requirements for products e. a protective glaze may be added to the frozen steak. 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. cod).g. A core temperature of around 27 30 ºC for approximately 100 minutes is a typical processing condition. or pre-packed in MAP or vacuum packaging (VP). The pâtés are sold from segregated ‘wet fish’ counters as a ready-to-eat product.g. The expected shelf life of these products is up to 8 days (or longer for some chilled. Hot-smoked fish are prepared from brined. salmon. annatto with smoked haddock. and typically uses natural smoke passing over the fillets suspended on racks in a kiln (oak chippings.5% in the aqueous phase. swordfish steaks).2. Again. resulting in a product that is ready-to-eat without any further cooking. be flavoured with herb dips or peppercorns.g. tuna steaks) or band-sawn from a frozen block (e. 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. the ingredients premixed.2 days) longer than that of non-prepacked fish. or pre-packed frozen with added glaze. Cold-smoked natural fillets are prepared from a range of cold water fish including herring.5 . Fresh fillets are sold from wet fish counters. They are then kilnsmoked to a minimum core temperature of 66 ºC for 2 minutes. 29 . natural fillets of fish such as herring. haddock. with a minimum residual salt content of 3. They are sold from wet fish counters. VP products) because of the high salt content acting as a barrier to spoilage. A typical cold smoking process may pretreat the fillet by dipping in a 26% brine solution with a retention time of approximately 3 minutes. mackerel. Pâtés are prepared with raw fish. etc.g. The shelf life for MAP fillets is typically up to 5 days. and salmon. This process in effect cooks the product and reduces or eliminates microbial contaminants.CHILLED AND FROZEN PREPARED FISH PRODUCTS frozen water that forms a protective glaze. Some products may be salted for flavour only (e. which may.5%. The result is fish with a typical residual salt content of 1. Cold smoking is essentially a flavouring process. Fish steaks can be cut from a natural fillet or loin (e.. in VP or through mail order. but may be increased (up to 1 month) by packaging methods like MAP or VP. in addition. sawdust with other woods adjusted for flavour profile and burn duration).
rice or pasta. pasta or potato. and formed into round cake shapes. long-life seafood mixes using sorbate and benzoate as preservatives.g. haddock. e. The cakes are usually coated using a three. mixed whitefish. Other. It is often used as a protein supplement. and may also include additional ingredients such as fresh herbs and spices. and in the shell. The protein source can be fish fillet or blocks. 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. salmon) coated and sold as chilled products.g. Natural prawns from cold or warm waters can include those peeled. jellied eels. Other ingredients may be natural or reconstituted. coated in batter or breadcrumbs.g. fish nuggets/bites with dips and seafood-based snack products. Fish cakes are products produced by mixing fish with potato and herbs. Scampi are tails of Dublin Bay prawn or langoustine.. Breaded fillets/steaks and battered fillets are either natural or shaped fillets. 30 .FISH AND SEAFOOD Seafood recipe meal centres (frozen or chilled) are places where these products are supplied without a source of carbohydrate. with an expected shelf life of around 7 days. There is an increasing market for high value fresh or thawed fish (e. etc. Prawns. it can be used as a seafood analogue. and may include molluscs or prawns that may or may not have undergone a minimal heat process during their primary processing. rice. addition of ingredients such as colours. taramasalata. and seafood mixes. frozen blocks or canned fish and may include molluscs or prawns that may or may not have undergone a minimal heat process. Surimi is the insoluble protein fraction of fish after washing and processing. ‘fingers’ before being coated in batter and/ or breadcrumbs. and heat-fixed. tuna. They are pasteurised products and treated as high risk raw materials. Band-saw dust and fish trimmings from fishblock preparation materials may be used. flavours.) Raw material is typically fish fillet. The majority are products made from various types of whitefish (cod. cephalopods. or cut from a preformed frozen fish blocks into shapes. e. With further processing. This ‘catch-all’ category includes seafood salads. which may include molluscs.or a five-stage process. binders.
and pathogenic flora associated with seafood. storage. see Table 2. skin.and turtlefriendly long lining to the less discriminatory netting. The Food and Drug Administration (FDA) has produced guidelines for handling and processing of tuna. product handlers. The concern is that fish may be visibly acceptable but any subsequent washing process will have little effect on removing contaminants. searing or bar marking). Only after death. 6). After capture and during any subsequent processing. as a result there is an increase in spoilage rate thereby reducing the product shelf life. Contamination will. i. Tuna caught and discarded as unsuitable for sushi or other high-risk uses is often diverted for canning. and his co-workers have described a large celled microorganism found in spoiling fish and identified it as Photobacterium phosphoreum (3).CHILLED AND FROZEN PREPARED FISH PRODUCTS 2.3 2. or visible damage from gaff marks (4). 2). For a summary of background information on the hazards associated with fisheries products and ingredients. sensory assessment of freshness etc. and have emphasised the origin and sustainability of the fish as key marketing points. whilst gaping is also dependent on a range of other factors that effect the muscle composition. specifying internal temperature limits for fish and for limiting temperature abuse during handling and grading (1). and intestines of the fish. it may be used for other products or markets depending on the product specification. Recent public opinion has favoured the use of dolphin. and an increased risk from a potentially unsafe product (especially those that receive a minimal heat process.e.3. the microbial flora of seafood is associated with the gills. and ingredients added to the final product. The edible tissue of some species of fish are more susceptible to gaping than others.1 Initial Microflora Seafood Other chapters have discussed in some detail the indigenous spoilage. fat content. remain on the surface of the fish fillet except where “gaping” or separation of the muscle blocks occurs. handling or preparation stage. other potentially pathogenic and spoilage bacterial contaminants may be introduced from a wide range of sources including the environment. which may become a focus of contamination deep within the muscle. on the whole. species (Bluefin>Bigeye>Yellowfin>Albacore). and processing conditions such as dehydration or poor filleting practices. In summary. Frequent rebaiting and prompt removal of caught fish is essential to limit rapid spoilage.I. Tuna that does not meet the highest of grades based on colour. will the saprophytic flora start to invade the surface flesh (1. including seasonal variation. A primary role in fish spoilage has been assigned to this organism. 31 . Grading is based on size of fish (>25 kg). is not used for sushi or other high risk (and high value) products. Dalgaard et al. and the potentially toxic by-products of histamine-producing bacteria (5.
Staphylococcus. and so are the potential microbial contaminants associated with them. nuts and fruit Associated microflora Lactobacilli.FISH AND SEAFOOD 2. Fusarium. shelf life determination. mushrooms. Enterobacteriaceae. streptococci. for chilled products. Escherichia coli. Clostridium Lactobacilli. moulds Bacillus. Bacillus subtilis B. intermediate processing. 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. coliforms. product assembly. cream. enteric viruses 2. by the HACCP team or by a hazard and risk assessment programme. Staph. cereus Bacillus licheniformis. Bacillus cereus. cereus Staphylococcus aureus. moulds Bacillus Bacillus. protozoa and viruses B. be considered at an early stage during new product development. cereus B. Moulds Enterobacteriaceae Salmonella. spore-formers. and milk Cheese Water/Ice Bacillus. 32 .4 2. TABLE 2. Clostridium botulinum. yeasts Lactobacilli. Aspergillus. Clostridium perfringens. Vibrio cholerae. histamine Cryptosporidium. Listeria. In addition. Table 2. and associated spoilage and pathogenic microorganisms. Penicillium. moulds Bacillus.3. pseudomonads. Pseudomonas. moulds.2 Non-seafood ingredients The range of ingredients used in the preparation of chilled and frozen prepared seafood products is very diverse. staphylococcal toxin. toxin. Aeromonas.4. Shigella. coryneforms.I provides an overview of common raw materials. anaerobic spore-formers. B. cereus Carbohydrates Potato flake and powder Rice Pasta Flour Dried herbs and spices Dairy products including butter. however. raw materials.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. 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.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. yeasts Pathogenic microflora/toxins Salmonella.
up to -5 °C so as to improve the cutting and handling properties during processing.CHILLED AND FROZEN PREPARED FISH PRODUCTS Any preparation stage or process should be suited to the raw material. Intermediate processing and use of chill for storage of work-in-progress will help to optimise available life of perishable components.3. as a heat process is often required for functional or organoleptic reasons. Examples of tempering techniques include the use of heated air. depending on the size of the fillet 33 . Generic examples have been used to illustrate only the major points.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.2. as the full complexity of the processes used for specific products cannot be covered in a general text.2 Raw material tempering Whenever frozen seafood is used as the raw material it is normally tempered before processing. line extensions and process reviews. Figures 2. but it also poses a risk of cross-contaminating other products undergoing tempering. Effects on the microflora are minimal during tempering (7). the core temperature. 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. but will also need to ensure that the optimum shelf life can be safely obtained. For some components.4. It is recommended that the principles of HACCP are applied to novel processes.4. but it is assumed that any band sawing or dice process will introduce heat from blade friction. Where the raw materials are frozen.5 illustrate basic principles in the preparation of chilled and frozen recipe dishes. Rapid controlled cooling of heat-processed components will help to minimise spoilage. and contamination from equipment to clean fish muscle. to evenly raise the temperature of the entire frozen product from -20 °C or lower. All have the same intention. or the use of microwave technology.1 . there are a number of distinct methods used. and potential opportunities for growth of foodpoisoning bacteria. 2. 2. defined holding time at a chill temperature (0 – 3 °C). this may be an incidental effect.3 Preparation of fish component of chilled and frozen recipe dishes 2. Band-sawn materials or fish dice are normally promptly further processed. It is critical that the time and product temperature during tempering be controlled to avoid elevated temperatures (>5 °C) on the product surface allowing microbial growth or the onset of spoilage in raw material.4. The use of water for rapid thawing of product is not recommended since it not only rapidly thaws the product surface.
1 (2.4. The “blown” meat is graded.4. 2. Individual Quick Frozen (IQF) fish fillets or steaks may also be coated using a similar process. by mincing and mixing with a binding agent (as described in section 2.2. 2.3. with the coating heat-fixed.7 days.2). Frozen cores are usually batter-enrobed. Any broken pieces or fragments of frozen tail meat are kept for use in the production of a reformed scampi product. On arrival at the processing plant. quickfrozen.1A .Shaped Fillets.3. frozen in a nitrogen blast freezer and weighed into sacks for later processing. may not rise above 0 ºC during processing.1. The processing of three examples types are described in Figure 2. drained and then refrozen for storage until required for further processing.Scampi. Fig.Fish Cakes).1.1. chilled or sold chilled on defrost with an expected shelf life of 6 . 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.1 Whole-fillet / scampi Scampi (Nephrops norvegicus). the scampi is washed in potable water to remove foreign bodies. 2. are placed into plastic boxes and covered in ice to preserve their condition.FISH AND SEAFOOD processed.1B . Raw material preparation stages for chilled or frozen coated fish products 34 . after landing. The scampi meat (tails) is removed intact from the shell using potable water whilst still frozen.1C . It is then size-graded.
those that use frozen blocks of whole fillets. 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. such as aerobic spore formers and Staphylococcus spp.off-cuts or belly flaps. spoil more rapidly than the dry ingredients. The fish component of the cake will. once the dry components have been rehydrated. as the physical structure of the material has been destroyed. iii) Fresh/fillet/minced fish block.4.3. or sold as a chilled product after thawing. The mix is shaped and formed ready for packing or for enrobing. Waste material produced from this type of processing is often reused in the third category (fish cakes). material will need to be transferred swiftly to a chiller. chilled.1. off-cuts and by-products from other processes (maybe minced) are reformatted give a final product that is still 100% fish. It is for this reason that all by-products from primary processing.CHILLED AND FROZEN PREPARED FISH PRODUCTS 2. fish fingers).3. and water is added to rehydrate the components.2 Shaped products This type of product falls into two categories.g. with an expected shelf life of 6 .4. or promptly processed to avoid further quality loss. This type of material will be susceptible to contamination by Enterobacteriaceae and possibly Staphylococcus spp. because of its nature. unless it is poorly handled. more seriously.. are handled and stored as any other food product. quickfrozen. This is especially the case with block-sawn dust. Fish raw material can be obtained from a number of sources i) By-product of fish shaping or cutting/dicing process . that are to be reused for other foods. with the coating heat-fixed. additional spoilage organisms may be introduced. Fish cake cores may be batter-enrobed.7 days. The remaining dried ingredients are mixed with the fish. Shapes may then be cut from the individual slabs using either hydraulic shape cutters or continued cutting with band-saw type blades (e. Waste fish generated as a by-product of another process will always be susceptible to spoilage and water loss. or those where the trimmings. from potato flake. ii) Fish-filleting waste . Spoilage of fillet pieces or trimmings will not be appreciably different from that of fillet.block-sawn dust.1. 2. but. pathogens such as Salmonella and Listeria. 35 .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. Herbs or seasonings used in the recipe may contribute additional spoilage organisms or. Therefore.
and Enterobacteriaceae from flour. The product then passes along a wire mesh belt through a ‘waterfall’ enrober that gives an even coating of batter to the entire product. quickfrozen.4 Battering/enrobing process Figure 2. a. Batter contains water. raising agents. rusk or crumb. which is product-dependent and not always required. Tempered band-sawn materials b. the crumb is poured onto the product followed by repeat enrobing and crumbing to complete the five-stage process.e. rapid automated processing with the minimum of delays will minimise product spoilage by pseudomonads. a characteristic flora will develop from the batter components. absorb any excess moisture on the surface of the fish portion.g. lactobacilli from the milk powder and flour.g. chilled. and if required. e. flour. 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. Bacillus spp. The first stage. fish cakes Portions or fillets may be batter-enrobed. because of the intensive handling required during processing. A large variety of product formats and types can be batter-enrobed.4. from flour. is the addition of a pre-dust. Breaded or battered fish fillets are fully heat-processed before consumption. The function of this is to improve the adhesion of subsequent coatings.. with an expected shelf life of 6 . Scampi tails are particularly susceptible to contamination with Enterobacteriaceae. and may include milk powder. or sold as a chilled product after thawing. perfringens could. which typically consists of flour. Control of temperature limits the potential for growth of spoilage organisms and pathogenic bacteria. Products are preformed and presented to the process (either three. starch. egg. e. Spore-forming anaerobes such as C. such as Listeria and Vibrio spp. milk powder and spices. with the coating heat-fixed. potentially. and possibly Staphylococcus spp. some of this initial flora 36 . 2.7 days. If a crumb coating is required. i. Natural fillets d. and to carry flavourings where required.2 illustrates the typical stages of enrobing or breading. spices. as well as ensuring that there is minimal contamination from human processors. rapidly multiply if strict temperature control was not maintained. vibrios and other fishassociated spoilage organisms. Initially. Fish products. Over time. IQF/pre-formed fillets c. any excess is removed with an air knife. which can occasionally be isolated from protein-handling processes.FISH AND SEAFOOD In all of the above examples.or fivestage) frozen or chilled.
Staph. and both of these can produce heat-stable toxins that can survive a batter heat-fix process. The origin of contamination for the two organisms is often unclear. Fig. It is advisable that some control mechanism is employed to prevent toxinforming organisms from multiplying to dangerous levels. time-limitation or production run temperature control. The FDA has issued clear guidelines on acceptable 37 . dry batter mixes are all potential sources. replaced in part by process contaminants and bacterial flora from seafood. This can be achieved by frequent clean-downs and adequate hygiene. contaminants such as Enterobacteriacae can proliferate and develop during the process. Flow diagram outline of a typical three. and. in the case of B. but it is believed that poor hygiene regarding product and food handlers. With inadequate temperature control of batter systems. cereus can proliferate under certain conditions. this is not of great significance to the microbiological standard of the finished product. aureus and 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. but because batter is heat-set.2. cereus.2.
For example. 2.5 Recipe products These products involve the combining of seafood with other food ingredients as part of a recipe. rice. Vegetable quality is an important characteristic often overlooked in the selection of vegetables for frozen and chilled products. but they are supplied without a source of carbohydrate e.g.4. The protein source can be fish fillet or block. Flow diagram outline for recipe dish ingredients preparation 2. to produce a specific product. as this could introduce off-flavours and reduce shelf life. The vegetables must withstand processing. 38 . and must not show evidence of disease. and may include molluscs or prawns that may or may not have undergone a minimal heat process during their primary processing. Figure 2.3 illustrates vegetable processing and treatment for inclusion in chilled and frozen fish recipe dishes. canned fish may be used in their preparation owing to it having wider availability and low wastage.3. A temperature no greater than 10 ºC for 12 hours cumulatively or 21.1 ºC for no more than 3 hours is recommended (1).5.FISH AND SEAFOOD temperature regimes and run times permitted for batter enrobers.1 Seafood recipe meal centres (frozen or chilled) These products can be defined as the central component of a meal. Fig. pasta or potato. Well-managed breading and pre-mix systems that are segregated from batter enrobers present little hazard (7). There are two main categories.2.4.
This is a concern for chilled products that may have a shelf presence for up to 6 days including day of manufacture. Blanching of vegetables can be an effective means of reducing or removing surface contamination. they should be stored under temperature-controlled conditions. pastry. For example. control is more onerous. all-year-round fresh vegetables are preferred. and may include molluscs or prawns that may or may not have undergone a minimal heat process. and mixed whitefish). larger diced pieces allow for easier handling and a reduction in spoilage but are less attractive to the consumer. Obvious signs of vegetable soft rot due to mechanical damage or infection by Erwinia spp. with the accompanying release of additional nutrients by processing.CHILLED AND FROZEN PREPARED FISH PRODUCTS The products will often be presented with vegetable toppings. coryneforms. coli O157. Once vegetables have been blanched. 8) will.3 log reduction in bacterial numbers on the surface of vegetables.e.lactobacilli. However. chopping and mixing of material. with cases of foodborne disease traced to Listeria monocytogenes.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. not just as accompaniments. and protected from any post-processing contamination. Any assumptions regarding the application of good agricultural practice to produce should be made with caution. The presentation of vegetables has a significant effect on the product quality. acidification of hypochlorite and chlorine dioxide have been proposed and recommended as an improved alternative. colour and water will occur. Salmonella typhi. Although the processes are claimed to be superior to using unmodified hypochlorite for washing. spore-formers. will disperse the saprophytic flora of vegetables . For example. and may include sauce accompaniments. 39 . or breadcrumbs. and reduce spoilage organisms and potential pathogens. The opportunity for growth of L. frozen block or canned fish. For premium products. A number of examples of direct faecal contamination of vegetables exist in the literature (8). if the process is not appropriately controlled. encourage more rapid decay and loss of structural integrity. 2. coliforms and pseudomonads throughout the mix.5. rice or pasta. haddock. even those with pH-modified dressings. any processing stage. Products must be washed thoroughly to remove pests and soil. The majority are products made from various types of whitefish (cod. treated as a highrisk ingredient. loss of quality. Washing with a biocide will at best achieve a 2 . and E. the use of night soil or incorrectly applied manure or sewage sludge for ground preparation may contaminate produce with a wide range of pathogens. but also as components in recipe meals. i. Raw material is typically fish fillet. In an attempt to improve washing and reduce chlorine taint. However. monocytogenes should be taken into account when setting shelf life or holding times of vegetable components. for example. (7.4.
4. B. such as butter garnish with black pepper. and then cooled slowly allowing germination and re-growth. can germinate and rapidly multiply. Chilled pasta is becoming more popular than dried pasta as a component in many product lines. such as high-risk/low-risk barriers and dispensing and/or holding at above 63 ºC. Sauces can be multi-component preparations or more simple preparations. aureus toxin is well documented (10. if subjected to temperatures between 20 and 40°C. should be chilled to <5 °C as quickly as possible using either blast chillers or other equipment specifically designed for this purpose. cereus. with the range always expanding as new tastes and cuisines are developed. Alternatively. or use barrier or hurdle principles to preclude the growth of pathogenic microorganisms. a process equivalent to at least a 5D Listeria kill will be necessary. An incident involving Staph. Manufacturers of chilled products with an extended shelf life must recognise the potential for growth and spoilage of psychrotropic strains of B. On hydration. Typically.6 Preparation of non-fish components of chilled and frozen recipe dishes 2.g. 40 .1 Sauce manufacture The variety of ingredients and components used in sauce manufacture is extremely wide. but temperature-abused pasta can also spoil rapidly by mould growth. rely on the use of microbiologically sound ingredients.6. cereus are well known and are still a common issue in the UK (9). 11). 2. For this reason all products. Placing such products in a standard chill store to cool down could result in condensation. a heat process in excess of 73 ºC will be required to rehydrate this component fully. Sauces or marinades are added to fish to broaden appeal or to accompany components.4. cereus). where composite meals are heat-treated. Potato toppings made from flake or powder will normally have little associated spoilage flora. the preparation process must ensure the eradication of all vegetative pathogens from the ingredients. 13). Products or dried ingredients contaminated with either spores of aerobic bacteria (e. monocytogenes and other bacteria that may be present on chilled surfaces such as ceilings.) to the product (12. spore-formers can proliferate if adequate control of temperature is not maintained. and an increased risk of contamination from L.FISH AND SEAFOOD Food-poisoning incidents traced to rice and caused by B. Spices and herbs added to form toppings along with breadcrumbs or other coating materials will similarly contribute a spore loading (primarily Bacillus spp. especially in those products that have a vegetable component or rice accompaniments. If the sauce contains modified starches for freeze/chill stability. Appropriate handling facilities or equipment will be required to protect the product. This is a particular issue especially where products are heat treated to eliminate vegetative cells.
Figures 2.herbs. 13). etc. respectively. 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. Marinade-type products are becoming more popular.4 and 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. and handling of processed sauce can be less rigorous than. Without barriers. Flow diagram outline for recipe dish ingredients preparation Time limitation may minimise the potential for spoilage by process contaminants or at the dispensing unit. for instance.4. suspended in an acidified sauce with an oil component.5 illustrate the recipe dish ingredients preparation and assembly and packaging. It is expected that this product format would have a restricted shelf life to limit potential for growth of psychrotrophic bacilli or any survival of enteric pathogens (12. Acidified or low-pH sauce formats using acetic acid or vinegar normally require a lesser heat process for the equivalent effect.2. 41 . spices. flour products. The format uses flavour compounds .CHILLED AND FROZEN PREPARED FISH PRODUCTS Fig. that of a dairy-based sauce.
4. which may restrict either microbial growth or enzyme activity.7. so replacing the oxygen necessary for aerobic growth of spoilage microorganisms. 2. unless the temperature is strictly controlled to below 5 °C throughout the supply chain. The objective of this method is to deny the aerobic bacteria (both spoilage and pathogens) the oxygen that they need to grow. as it dissolves in the surface moisture to form a weak acid. generally carbon dioxide and nitrogen.2 Modified-atmosphere packaging Originally developed for the red meat industry.FISH AND SEAFOOD Fig.7.7 Effects of packaging on chilled products Products with minimal additional ingredients may use packing techniques designed to minimise microbial growth. These include: 2.4.5. Flow diagram outline assembly and packaging of recipe dish 2.1 Vacuum packing A range of fresh chilled products including salmon (smoked and fresh portions). 42 . the method replaces air in the sealed packaging with a defined mixture of gases. The oxygen-free conditions will however allow other anaerobic bacteria such as C. and tuna loins are packed by placing the product in a plastic pouch and removing all the air.2. Increased levels of carbon dioxide will also result in a slight decrease in the pH of the surface tissues.4. botulinum to grow.
initially.CHILLED AND FROZEN PREPARED FISH PRODUCTS 2. It is not clear whether the additional contaminants from components or processing will accelerate this typical seafood-type spoilage. Spoilage will be most readily detected in proteinaceous components of ready meals. phosphoreum. and finally flavour loss. by a loss of texture. temperature control is important in preserving chilled integrity of ingredients.5. This is also true of chill storage and the distribution chain. Nonetheless. both organisms are capable of growth at chill temperatures. The fish component of recipe dishes.1 Chilled storage For chilled products or chilled components of frozen products. but little on in-pack spoilage. (20). and in influencing the final eating quality of the product. where the protein component is nearly always heat-processed. However. at slightly higher temperatures (10 ºC). This is considered to be the one major difference between fish and meat products. advanced spoilage gives further changes in texture and the production of characteristic ‘off-odours’. 43 . is typically uncooked or at best lightly processed to retain eating characteristics. The case for growth and contamination of products with Aeromonas is stronger. in numerical terms. as this organism is a common contaminant of freshwater fish species (19). Shewanella putrefaciens and Ph. a large number of nutritionally non-fastidious spoilage organisms are commonly isolated from seafood. Aeromonas spp. followed by moisture loss. but they will also be readily adapted to the nutrient source and conditions. Not only will these organisms have a numerical advantage over other proposed spoilage organisms. but a conclusive case for pathogenicity has yet to be presented for Aeromonas spp. the slow development of ammonical and sulphuric ‘off’ odours and flavours(16). except for some crustacea. The literature has good information on spoilage of individual components of recipe meals.. the microflora of interest will be mainly psychrophilic or cold-adapted (14) spoilage organisms such as Pseudomonas spp. Spoilage of this material is not expected to differ greatly from the processes described elsewhere. and Yersinia spp. Vibrionaceae will succeed (17 ). but is not limited to.5 Spoilage During processing. Beyond this. have both been suggested as possible spoilage or pathogenic contaminants of chilled recipe meals (18). At refrigeration temperatures (5 .10 ºC) Enterobacteriaceae are often found to proliferate and produce a characteristic type of spoilage that includes. and therefore will have a greater opportunity to dominate other spoilage flora numerically over the course of product shelf life (15). 2.
however. For example. This reinforces the need for prompt processing and temperature control of ingredients and finished products. botulinum. freezing also has a protective role in improving the safety of raw materials by.. a recent report examining fish gut bacteria from cold waters characterised a proteolytic enzyme. and Aerobacter spp. The conditions and freezing process most favourable for good-quality products will.FISH AND SEAFOOD 2. Serratia spp. where the variety of components and ingredients used in manufacture is varied and complex. Products that are developed with an extended shelf life beyond the 6 . released by a pseudomonad with demonstrable activity to 0 ºC and a peak in production at 10 ºC (15). also be those that have the least effect on the viability of microorganisms (21). Still. for example. eliminating nematodes in products that do not have a heat treatment step. Characterisation of the enzymes has demonstrated residual activity even after a heat process that would destroy vegetative cells (24). the shelf life of products is limited by the natural spoilage process described earlier. These have been proved to impart off-flavours. this should also take into account shelf life of product components used in the assembly of the final product.3 Enzymic spoilage Post-harvest changes in the quality of fish due to enzymic activity and rigor mortis are well known and understood (10). 2.5.7 days typical of fish products/recipe meals must consider the potential for growth of non-proteolytic C.g. The type and nature of the enzymes and their activity will depend on the bacterial species. The ACMSF (22) issued clear advice on this topic and has recommended that the shelf life of products without barriers (23) is restricted to less than 10 days.. auto-oxidation of fats/oils). enzymic processes are often ignored or overlooked as insignificant or unimportant because of the rapidity of fish spoilage. within dairy products are well known. producing characteristic off-flavours. Bacillus spp. spoilage should not be considered a purely microbial phenomenon as taints and off-flavours can develop by non-microbial means (e. As a note of interest. 44 . However. Pseudomonas spp.. Other examples of bacterial proteolytic and lipolytic enzymes produced by Alicaligenes spp. This also ensures that any post-process bacterial contamination or spore-forming bacteria in the product will not grow. It is not unreasonable to expect that spoilage bacteria release many extracellular enzymes. Spoilage can be either a function of bacterial growth or the microbial release of free enzymes into solution. By retaining the intrinsic spoilage flora of seafood. which would warn the consumer of the potentially unsafe nature of the product.5.2 Frozen storage Maintenance of product temperature at <-18 ºC will not permit bacterial growth or enzymic spoilage.
These are C. are dealt with elsewhere in this book. A number of other Bacillus spp. potato.CHILLED AND FROZEN PREPARED FISH PRODUCTS 2. soil. Vibrio spp. Additional hazards from marine bio-toxins. In addition. farmed or free-living crustacea (30). botulinum type E and Vibrio parahaemolyticus (25). Hazards attributable to the non-seafood components are listed below and this information. typhi. perfringens contaminating seafood will depend on the source of raw material. Numerous quoted figures exist in the literature regarding cell counts required before toxin production occurs. and this issue may be of greater consequence for warmwater. 45 . water. low oxygen or anaerobic conditions.1 Bacillus cereus This is a common contaminant of dust. permit rapid growth of B.6. spices and herbs and vegetable shoots have been reported (12. histamine intoxication.. Inappropriate holding times and temperatures will. especially molluscan filter feeders. with pathogenic microorganisms or viruses.e. air. may be of benefit within any risk assessment for chilled and frozen prepared fish products. etc. 2. Therefore. 2. and some effects related to product type and time to toxin production have been noted. botulinum when held in favourable conditions for growth (28) i. Incidents in such diverse materials as cream.2 Clostridium botulinum There are reports of rapid spoilage of vegetables by C. along with previous discussion. perfringens in seafood noted a transient population of spores in the guts of fish feeding adjacent to rivers and sewage outfalls (29). Counts of between 103 . 2. in some cases.104/ g should be treated with some concern (26).3 Clostridium perfringens A study examining C. and many raw or processed foods (26). Cases quoting contamination of seafood with organisms such as S. agricultural run-off or sewage can contaminate in-shore species.6 Pathogens: Growth and Survival There are only two species of pathogenic bacteria that can truly be said to occur naturally in fish. cereus. Germination of spores requires favourable conditions.6. any risk of C. This may occur where storage under gas such as carbon dioxide is used to reduce ripening and so prolong storage. and Campylobacter are recorded in filter feeding molluscs such as mussels. 27). 13. or a heat shock followed by holding at a temperature suitable for growth.6. have been linked with food poisoning and should also be considered.
but under the appropriate conditions. aureus enterotoxins in foods. monocytogenes as a contaminant in foods (31). indigenous or other.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). Chilling may help Shigella to survive. 2.6.FISH AND SEAFOOD Vegetable raw materials are potentially contaminated through soil conditioning with animal manure.6. Shigella spp. All the above points should be considered within any risk assessment of the potential for growth and toxin production by Staph. An acceptance has now developed in the UK of the presence at low levels of L. These include inadequate refrigeration and preparing foods far in advance of planned service.5 Shigella spp. 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). and this is believed to be due to a tolerance to brine used in the smoking process (32). L. which is not affected by any subsequent cooking process.6 Staphylococcus aureus There are a number of underlying reasons for the development of Staph. monocytogenes has been frequently isolated from fish-processing plants and is thought to be able to colonise this type of environment (32. and will therefore cause “food poisoning” even in a product that is cooked before consumption.6. 33) due to its ability to grow at low temperatures and form “biofilms” on surfaces. growth may occur in foods (26). 46 . when introduced or inoculated into sterile components of chilled prepared recipe dishes. will colonise factory environments in a similar fashion except for possibly Vibrio spp. However. the restrictions on growth rate seen with seafood will not apply. and inadequate cooking or holding within the temperature growth range (27). the possibility of a sanitiser-resistant biofilm colonising processing environments should be viewed with some concern (32). if any other potentially pathogenic organisms. monocytogenes tends to grow poorly at refrigeration temperatures in seafood (33). 2. These conditions will allow the production of the toxin. Sources of contamination are many because of the widespread distribution of this organism in the environment. L. Rates of contamination with smoked seafood are much higher than those with unsmoked products. It is unclear. at the present time. infected food handlers. but. 2. aureus (35).
with respect to the number and types of ingredients used. and as such the monitoring is for verification purposes rather than control. 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. the Institute of Food Science and Technology (IFST) (39) or the British Retail Consortium (BRC) (40).III based on IFST Guidelines (39)) provide example guidelines for seafood products. (36). and are a recognised means of providing data on manufacturing standards. but these may be eliminated by a subsequent heat-treatment stage. Microbial guidelines. pasta. cereus emetic toxin are often confused owing to similar symptoms. such as batter mixes. pastry.II and 2. as published by for example.7 Published Microbiological Criteria Due to the complexity of many processed food products. In the EU these are defined under European Commission (EC) Regulation 2073/2005 (38).6. are not mandatory. and not to final products. For instance. Microbiological specifications do not necessarily relate to product safety. the fresh product is often consumed before the results are obtained. and their association with similar materials. . 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. it is not surprising that Microbiological Standards (defined as mandatory compliance required (37)) are limited to those raw materials in which specific hazards occur. although the only seafood products specifically identified are raw “shelled and shucked products of cooked crustaceans and molluscan shellfish”.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. 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. aureus enterotoxins and B. etc. 2. Microbial criteria and the monitoring of microbial levels within a product are important for monitoring trends within production systems. although due to long analysis times required. 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.A dairy-based sauce that includes a non-pasteurised cheese will contribute significant aerobic total counts. The following tables (Table 2. 2. It should be remembered that 47 .CHILLED AND FROZEN PREPARED FISH PRODUCTS Illnesses caused by Staph.
fish fingers. ready meals Storage: Frozen/chilled Use: Re-heated prior to consumption Pathogens and toxins Organism Salmonella spp. TABLE 2.fish products.g. 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. L.III IFST Guidelines for processed foods .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.II IFST Guidelines for part-cooked foods – e. parahaemolyticus (warm-water fish) Histamine (scrombroid fish) Indicators and spoilage organisms Organism APC (heat treated) Enterobacteriaceae E. cereus Staph. aureus V. monocytogenes B. coated fish. coli GMP* <102 Maximum 104 TABLE 2. Maximum= Levels at end of stated shelf life 48 .
. Masuoka Y. Yumoto I. 49 4. 1997.T.. Powers E. 12. Journal of Milk Food Technology. 1997. 2.G. 14. Sakamoto T. 1962. Murrel H.. London. International Commission on Microbiological Specifications for Foods.R. 59 (2). Bacterial histamine production as a function of temperature and time of incubation.. Eds. Martin R. Matsuyama H.K. 9. References US Food and Drug Administration. Communicable Disease Report... 13. Hackney C. The bacteriology of fresh and spoiling fish and some related chemical changes. Fish and Fisheries Products: Hazards and Controls Guidance.C. Isolation of a Pseudomonas species from fish intestine that produces a protease active at low temperature. Animals and Spoilage.L.W. Tu A. 39 (10).. Ohgiya S. Christiansen T. Fish: microbiological spoilage and safety. Microbiology of Marine Food Products.. London. Behling A. 11. 7.. 1997. 663-87. 1311-7. Microbiology of processed spices.. Food Science and Technology Today. 1993. 1976. Shewan J. 1. Incidence and levels of Bacillus cereus in processed spices. Microorganisms in Foods 6: Microbial Ecology of Food Commodities. 2. 1991.H... 1997. Dalgaard P. Lawyer R. 3. Woolaway M. Collette R. 373-8. Beuchat L.. Proper processing and handling of raw tuna. 38 (11). Letters in Applied Microbiology. Recent Advances in Food Science. in Fish Inspection Quality Control and HACCP: A Global Focus. 5. Gilbert R. Outbreaks of foodborne illness in humans..M..E. 15.. Pathogenic microorganisms associated with fresh produce. 67-73.R.A. Pvt. Ishizaki K. Mejholm O. Wienke A.. 70-2. IBH Publishing Co. Ltd. 1976.. Hobbs G. 167-93. Latt T.J.L. Ward D.. Communicable Disease Report. 1986. 10. Toxin Related Diseases: Poisons Originating from Plants. Kluwer Academic. Cambridge.. Oxford. 166-73. 8.M. ..R.. Washington D.R. Importance of Photobacterium phosphoreum in relation to spoilage.C. Bartram P. Journal of Food Science. Van Nostrand Reinhold. Aureli P. 6. Hoshino T. 215. New York. 2005. 25 (1). 96. 2001. Food and Drug Administration. Journal of Hygiene. Taylor S. 668 -70.CHILLED AND FROZEN PREPARED FISH PRODUCTS An authoritative survey of specifications can be found in Shapton & Shapton (41).M.8 1.C. 7 (24). Journal of Milk Food Technology. 24 (5). Kumeta H.R. 204-16. Letters in Applied Microbiology. 1982. 1991. Slavin J. Taylor and Francis. Journal of Food Protection. Huss H.L. Bartlett C. 5 (3). 233-7. England and Wales Quarterly Report. International outbreak of staphylococcal food poisoning caused by contaminated lasagne. Powers E. Brown T.J. 47.
274-86.A. Food Taints and Off-flavours. 1987. Matches J.. What problems does the food industry have with spore forming pathogens B. 1986. Leatherhead. 25 (2).. 1992. Gram L. Food Control. 1996. Speck M. 1977. Jemmi T. Ray B. Mead G.M.. 24.K. 18. coli (VTEC) . Blackie Academic Press. 20. 19. Journal of Food Protection. Golden D. 51-158 Saxby M. SAB Symposium Series No.. 11. 28. 29. Huss H. 26. Liston J. Anaerobes in relation to foods of plant origin. Curran D. New York. 4 (1).L. 40 (5). Anon. Modern Food Microbiology. Leatherhead Food Publications. a foodborne pathogen? Review of the clinical data. in Advances in Fish Science and Technology. Granum P. PHLS Microbiology Digest. 30.. Journal of Food Safety.W.H. 1995. 655-60. 59-72. 155-9. International Commission on Microbiological Specifications for Foods. 1992. Keusch A. Gram L.C.. Report on Vacuum Packing and Associated Processes. 50 17..C. Eds. 28 (4). 28. London.. Microorganisms in Foods 5: Microbiological Specifications of Food Pathogens.. Trolle G. 9. 1993. Wood L. Food preservation by combined methods. 1996. Applied Microbiology. 65. 4. London. Advisory Committee on the Microbiological Safety of Foods.. Microbiological guidelines for some ready-to-eat foods sampled at the point of sale: an expert opinion from the Public Health Laboratory Service. Ed. 1988.C. 13 (1).A Literature Survey. in The Microbiological Safety and Quality of Food. Baird-Parker T. Is Aeromonas spp. Springer. perfringens? International Journal of Food Microbiology.M. Gould G.. 351-72. Leistner L. International Journal of Food Microbiology.J. Survey of the incidence of Aeromonas and Yersinia species in retail foods. Aspen Publishers. Effects of freezing and storage on microorganisms in frozen foods: a review. 32. Occurrence of Listeria monocytogenes in fresh water fish farms and fish smoking plants. cereus and C.J. 34-40. 25. 2005.Yersinia. Walker S. The Emerging pathogens . Andersson A. 13.E. 472-506. 333-6. Detection of specific spoilage bacteria from fish stored at low (0 ºC) and high (20 ºC) temperatures. 31.J. Connel J. Food Research International. Lund B.. Halligan A. Liston J. Her Majesty’s Stationery Office.J. Blackie Academic and Professional. 1994. 1996. Microbiology in fishery science. 1974. Clostridium perfringens in the environment. Jay M.. Blackwell Scientific Publications. .. U. 1979.FISH AND SEAFOOD 16. 41-3. 21. 23. 309-16. Brooks J.R. 1990. Food Microbiology. 2000. Aeromonas and Verotoxigenic E. Morgan D.. Gaithersburg. Oxford. Eds. Loessner M. Lund B. 22. Glasgow.V.R. Barnes E. Ronner U. Fresh and processed fish and shellfish . in Anaerobic Bacteria in Habitats Other than Man. Huss H.H. 27. Fishing News Books Ltd.J..
chilledfood. Kraft A. Chilled Food Association. U. Wolfe Publishing. 37.K.2. Institute of Food Science and Technology. British Retail Consortium. 1997. U.europa. London. Shapton N. 34.uk/cday/posters2008/npd6. 38. Gloucestershire.. 39.eu/LexUriServ/ LexUriServ.CHILLED AND FROZEN PREPARED FISH PRODUCTS 33. Incidence and survival of Listeria monocytogenes in ready-to-eat seafood products. 50 (9). http://www. Principles and Practices for the Safe Processing of Foods. 372-6. London. CRC Press. Campden & Chorleywood Food Research Association.co.org/Resources/Chilled%20Food%20Association/Public%20 Resources/BRC_CFA_Micro_Criteria_Guidance_Ed_1. 60 (4). 1991. Cambridge. Chilled Food Association. 35.G. 51 . 1998. Guidance on the Practical Implementation of EC Regulations on Microbiological Criteria for Foodstuffs. Shigella as a foodborne pathogen. Woodhead Publishing Ltd. http://eur-lex. 788-801. Evans M. Development and Use of Microbiological Criteria for Foods. 2006.K. Varnam A. Journal of Food Protection. 36. Foodborne Pathogens an Illustrated Text.pdf Commission Regulation (EC) No 2073/2005 of 15 November 2005 on Microbiological Criteria for Foodstuffs. 1999.. 1992.A.L. Phil Voysey. 40. http://www.A. Smith J.F. Microbiological Criteria. McCarthy S.pdf Shapton D.H. Psychrotrophic Bacteria in Foods: Disease and Spoilage. 1987.campden. 41.A. Journal of Food Protection.do?uri=CELEX:02005R2073-20071227:EN:NOT Institute of Food Science and Technology.
The class having species of economic and public health importance is the Pelecypoda. viruses. chemical contaminants and other impurities on the mucus of the gills. The digestion process requires less than 2 hours in actively feeding adult bivalves (2). cockles. cockles. where it is eventually swallowed and digested. lying well within the mantle cavity. Box 3012 Bothell WA 98041-4421 United States of America 3. cockles and scallops. Other species of shellfish such as crabs. Scaphopoda. The term shellfish in this context is limited to oysters. snails. P. O. Food and Drug Administration 22201 23rd Dr. and tooth shells are a group of soft-bodied animals that usually secrete external protective shells. can hold the shells tightly shut against enemies (1). oysters.1.1 Definitions General features of the mollusc The phylum Mollusca. abalones. MOLLUSCAN SHELLFISH Carlos Abeyta. and Cephalopoda. In the process of filtering water. composed of two valves hinged together dorsally. In these forms. Pacific Regional Laboratory Northwest U. the foot is compressed to form a muscular spade for digging and the head is greatly reduced. Many microorganisms ingested by shellfish survive the digestive process (3. and scallops. The phylum consists of five classes: Amphineura. lobster.3. slugs. which includes oysters. The class Pelecypoda includes all of the bivalves such as clams. and shrimp. respectively. oysters and clams can process about 3. mussels. squid. Pelecypods have little need for locomotion. they also trap bacteria. In most species. mussels. Plankton and algae form the bulk of their diet from seawater. octopi. clams. while 53 .S. Jr. the shell. 4). For example.1 3. chitons. the adductors. Pelecypoda. S.E. and 2. clams. scallops. Special tracts of cilia move this mucus toward the mouth. mussels. Strong muscles. whelks.6 litres. Gastropoda. completely encloses the body. Bivalves feed by filtering large volumes of water across their gills to obtain oxygen and food.7 litres of water per hour.
Mactra sachalinensis Saxidomus giganteus Japan Pacific – North American Saxidomus nuttali Protothaca staminea or Paphia staminea 54 .FISH AND SEAFOOD commercially valuable.South America Pacific – Japan Quahaug (also known as hard clam and hard shell clam) Coquina clam Hen clam Butter clam (also called the Washington clam) Mogal clam Little neck clam (also called rock Cockle Pacific littleneck) Atlantic – North American/Europe Atlantic – Europe/North America Japan Atlantic/PacificNorth America Donax spp. dwelling on the bottom or attached to structures in the water column. and usually at higher concentrations than found in their habitat.2 International commercial products Globally. Whatever contaminants are in the water will eventually get into the shellfish. are not as vulnerable to contamination through pollution of their beds as the bivalves. 3.I Partial list of bivalve molluscan shellfish products in commercial use internationally (5) Common name Clams Soft (shell) clam Pacific/Atlantic North America Pacific/Atlantic North America Pacific .North America Saxidomus nuttali Venus mortoni Protothaca thaca Meretrix spp. Clams are benthic and live burrowed in the mud.I). many species of bivalves are edible (Table 3.1. The following are descriptions of the various bivalves and their forms of preparation for human consumption (6): TABLE 3. but maintain contact with the water by means of a siphon tube (5). The habitat of oysters and mussels are sessile. Mercenaria mercenaria Venus mercenaria Mya arenaria Country Scientific nomenclature Hard clam (also used for Quahaug/quahog) Pacific .
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.
2. hard-shell or soft-shell clam. littleneck.1. and other names. Buried clams leave tubes. The muscular foot is used to burrow in mud or sand.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. cherrystone. Jacques Iceland scallop Bay scallop Weathervane scallop (also called Alaska scallop) Atlantic North East Atlantic North Atlantic West Atlantic Pacific – North America Pacific – North America Pacific – Japan Common scallop Sea scallop (also called giant smooth) Japan Atlantic – North America Pecten varius Pecten maximus Chlamys islandica Argopecten irradians Pecten caurinus Pecten aequisulcatus Pecten laquaetus Pecten yessoensis Penten magellanicus or Placopecten magellanicus Pecten meridionalis Pecten novaezealandiae Pecten jacobaeus Aequipecten gibbus Chlamys opercularis Chlamys varius Amusium balloti Mediterranean – South East Europe New Zealand Australia Mytilus galloprovincialis Mytilus canaliculus Mytilus planulatus Commercial scallop Australia New Zealand Great scallop Calico scallop Queen scallop Variegated scallop Scallop saucer Atlantic/Mediterranean Atlantic – North American Atlantic Atlantic/Mediterranean Australia 3. extended above the surface to maintain water currents needed for respiration and feeding.FISH AND SEAFOOD Mussel Mussel Mussel Scallops Scallop Coquille St. and two symmetrical shells (valves) held closed by two large muscles and joined by a dorsal hinge joint. Most clams are only a few centimetres in their maximum dimensions. but 56 . geoduck. mostly marine bivalve molluscs. or siphons. laterally compressed.
and no siphon. clam madrilène and clam liquor. Smoked: meats. Canned: meats removed from the shell by steaming. then heat-processed. in cans with either brine or clam liquor. oyster bisque. or shelled meats. or shelled meats. Soups: oyster stew. boiled and then sun dried for 3 to 10 days before packing in boxes (Hong-Kong) Smoked: meats usually canned in edible oil. either whole or minced. 57 . Frozen: shelled meats. Live: in shell Fresh: in shell or shelled meats. These molluscs feed on minute plants and animals carried to them by the current. Canned: clams are steamed open and the meats removed from the shells. Canned soups include clam chowder.1. They live free on the bottom or adhere to stones or other objects in shallow water along the seacoasts. trimmed and packed.2 Oyster Oysters are marine bivalve molluscs. uncooked (shucked). sun-dried. Semi-preserved: meats cooked and packed in spiced vinegar.MOLLUSCAN SHELLFISH the giant clam of the Indian and Pacific oceans may be nearly 130 cm long and weigh 227 kg. having rough and irregularly shaped shells that are closed by a single adductor muscle. smoked meats are also canned. Dried: shelled meats. hermetically sealed. Fresh: in shell. skewered with bamboo sticks. oyster soup.2. the meats are smoked and packed in oil in cans or jars. washed. Oysters lack a foot or have a reduced foot. 3. Live: in shells. all in cans. Dried: Hoshigai (Japan) shelled meat. packed in weak brine. uncooked meats packed unprocessed. Frozen: in shell. after precooking and brining. uncooked. tidal waters or in brackish water in the mouths of rivers.
also with other ingredients. Canned: cooked meats. mussel in butter sauce.1. Live: whole with shells. composed of striated muscle fibres (used for fast swimming) and unstriated muscle fibres (used to hold together the two halves of the valves). by interlocking tufts of hair-like cilia.2. in mussel liquor.3 Mussel Mussels are marine bivalve molluscs that live either partially buried in the sea bottom or attached to rocky surfaces by means of byssus threads. The shells are fluted and round with a broad. sauces. Salted: cooked meats bottled in brine. or packed with jelly. are united.FISH AND SEAFOOD 3.1. They have filibranch gills.) Dried: peeled. scallops can swim or leap by a series of rapid closings and openings of the valves. and the anterior end smaller and pointed. 3. The common scallop is about 5 cm long. with the posterior ends of the shells (valves) broad and rounded. The posterior halves of the valves contain a large adductor muscle that holds the valves closed. Bait: used live extensively for baiting fish hooks for line fishing. They have a radially ribbed shell with the edge undulated and swim by opening and closing the valves. 58 . Semi-preserves: cooked meats with vinegar-acidified brine and spices. vinegar solution. brine. in which the individual branches. The adductor muscle is the only edible part of the animal. Unlike other sessile bivalve molluscs. boiled. Internally. Fresh: shelled meats (often only the adductor muscle and the roe are eaten. scallops possess a single large adductor muscle. mussel paste. Smoked: meats may be canned in edible oil. and held apart. gutted.4 Scallop Scallops are marine bivalve molluscs living in shallow waters of protected bays with sandy or muddy bottoms. The familiar marine mussel is asymmetrical. cooked meats packed in dry salt for transport. or filaments.2. In some species. the shell is often characterised with a lustrous nacreous or pearly lining inside their shells. flattened ear or wing at the hinge area. smouldered and afterwards dried (Japan). Fresh: cooked meats.
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. also bottled in brine. Salted: meats either lightly or heavily dry salted. and the environment. 3. and butter.2 Initial Microflora The microflora of molluscan shellfish directly reflects the environment from which the shellfish is harvested. Cockles range from about 1 to 15 cm in length and have two valves of equal size. nutrient concentration and pollutants of the water column.MOLLUSCAN SHELLFISH Frozen: shelled meats. Because molluscan shellfish are filter feeders. 3. Whatever contaminants are in the water will eventually get into the shellfish (7). which affects the temperature. mostly in oysters. are vectors of disease transmission (8). Canned: in own juice. Primarily inhabits shallow waters near tidal zones where they bury themselves in the sandy or muddy bottoms.1.5 Cockle A marine bivalve mollusc having a shell with convex radially ribbed valves.2. Vinegar cured: bottled in malt vinegar after brining. pH. The initial microflora of molluscan shellfish is comprised of the natural commensal microorganisms and the microorganisms accumulated from the water during feeding. Live: in shell. Raw and partially cooked shellfish. 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 . in sauce. salinity. clams and mussels. depending on length of journey. Fresh: meats removed from the shell by boiling. Canned: in brine. they accumulate pathogenic microorganisms from polluted waters.
Other microorganisms found in lesser numbers are the Achromobacter spp.3.2. clams.. 10. For example. Vibrios. Water temperatures affect the concentrations of microflora in the water. non-B Snow Mountain virus Plesiomonas shigelloides Salmonella spp.. Pseudomonas spp.1. Campylobacter coli Bacillus cereus Hepatitis non-A. dredges.3. and shucked shellfish that is shellfish.. 3.1 Shellstock 3. rakes. from which one or both shells have been removed. Flow diagrams of the major processing steps of molluscs are presented in Figures 3.1 Processing and its Effects on the Microflora Oysters. salinity. Moraxella spp. The microflora of molluscan shellfish at harvest consists predominantly of Gramnegative rods while the Gram-positives constitute a minor portion of the flora. spirochetes of the genus Cristispira and Saprospira.1. Enterococci spp. and Aeromonads. 12. At harvest. mussels. Campylobacter jejuni Escherichia coli Hepatitis A Norwalk virus Other viral agents A variety of commensal microflora are found in molluscan shellfish. or by 60 . 13). mechanical harvesters. Acinetobacter spp. pollution point sources and hydrological events will greatly influence the initial microflora in bivalves. (9. the initial flora composition and concentration of bivalves are highly variable. Micrococcus spp. Flavobacterium spp. There are microorganisms.3..000 /g.. whole or in part. 11. and yeast cells of Rhodotorula rubra and Trichosporon spp. Temperature. nutrients.. but have never been cultured (14.FISH AND SEAFOOD Aeromonas hydrophila group Staphylococcus aureus Shigella spp. Bacillus spp. They are termed viable non-culturable microorganisms.3 3. The largest groups found are the Pseudomonads.. shellstock (live) which is shellfish in the shell.1 Processing description The method of processing shellstock begins with the harvester collecting shellstock by the utilisation of tongs. molluscan shellfish have an Aerobic Plate Count (APC) of approximately 1000 to 100.1. In summary.. 15). have been observed microscopically in the digestive tract of Eastern and Pacific oysters.. 3. and cockles The processing of bivalves takes two forms. observed microscopically. that have never been cultured on laboratory media.1 and 3. responding to the environment in which they are harvested. Alcaligenes spp.
Shellstock may be wet-stored at the processing facility in tanks of clean seawater. There are three primary reasons for wet storage: first. or in bay-approved shellfish-growing waters prior to sale or processing. after harvesting. Generic flow diagram of live shellfish from harvester to point of sale 61 . Thirdly. or immediately placed in refrigerated trucks. usually in open-mesh containers and/or burlap sacks. Fig. Shellstock are given a wash to reasonably free them of bottom sediments and detritus as soon as practicable. or from other sources approved by the State Shellfish Control Agency (SSCA). 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. shellfish can be harvested from remote areas at convenient times and held live for brief periods at locations close to the point of sale. The shellstock is transported dry. by truck or boat. for transportation to the point of sale. Shellstock are placed in drycooler storage containers in the facility.MOLLUSCAN SHELLFISH hand (Figure 3.3. Water used for shellstock washing is from approved growing areas. wet storage may be used to increase palatability by increasing salt content of shellfish harvested from low salinity waters. If by boat.1. in containers from which they can be retrieved easily. Secondly. usually the boat will be docked by the processing facility with shellstock piled on the deck of the boat or dock.1).
24.FISH AND SEAFOOD 3. involving 691 cases was attributed to oysters contaminated in a wet storage area (27). coli. to avoid possible contamination of the shellstock. with prolonged storage the microbiological quality of the bivalve decreases. Wet storage operations are highly variable and may range from temporary storage near-shore in approved areas to on-shore tanks using re-circulating. Water used for shellstock washing should be of good sanitary quality. At this temperature the multiplication of faecal coliforms (E. Excessive sediment on the shells and dead shellfish may increase bacterial 62 . 22). bivalves tightly close their shells thus trapping water and microflora. Shellstock should be refrigerated at temperatures at or below 10 ºC (50 ºF). for a considerable length of time. There is a positive relationship between sewage-polluted shellfish and enteric disease transmitted by shellfish (8. with the exception of A. metabolic waste accumulates in the shell serving as nutrients for bacterial growth. vulnificus. and temperature storage of shellstock to point of sale. 21. without adequate refrigeration. the bacterial counts in the meats will increase. Several studies have established that pathogenic Vibrios and other bacteria may be present in marine sediments throughout the year (21. An outbreak of infectious hepatitis in Sweden in 1956. hydrophila (18). there is evidence that in the absence of adequate refrigeration. Shellfish may contain higher levels of pathogens than are found in the water in which they grow. shellstock are placed in refrigerated trucks for transport or placed in wet storage. Klebsiella. Enterobacter. The washing of sediment and detritus soon after harvesting will prevent further contamination.1. however. Shellfish in wet storage tanks are similarly subjected to pollution if the tank water is obtained from a polluted source. Several studies have established the increase of bacterial counts in shellstock during post harvest period before the shucking process (18. Furthermore.1. when it reaches the processing facility.2 ºC (45 ºF) or below (19). 17). Food and Drug Administration (FDA) recommends that potentially hazardous food such as shellfish be held at 7. including V. some pathogenic organisms will survive and multiply in shellfish (20. 23. are prevented. Generally. 26). 25). Harvest conditions are another factor that influences the growth of bacteria in shellstock. synthetic seawater for the purpose of de-sanding and salt uptake. Over a period. The safety of shellfish is predicated by the cleanliness of the growing area waters from which they are harvested. and the sanitary practices applied during harvesting and shipping. Washing shellstock helps prevent quantities of sediment and bacteria being mixed with. 16. At harvest. The most important factor in controlling bacterial growth in shellstock is temperature. handling. Oysters will remain alive in the shell for approximately two weeks if kept at the recommended temperatures. shellstock are not refrigerated on the harvest boat.3. and Citrobacter species) and Vibrios.2 Effects of microflora during processing The initial microflora found on or in shellstock is influenced by sources of shellstock. Generally. If long delays occur before reaching the processing facility and/or refrigeration. and thus contaminating the shucked meat at the point of sale.
MOLLUSCAN SHELLFISH loads in the tanks and lead to increased microbial levels in the shellfish during storage.1. and packing shellfish into containers (Figure 3.2. lobsters. Shucking and washing of meats under sanitary conditions prevents the proliferation of bacterial populations. washing and culling shellfish prior to storage is essential. shellstock is subjected to contamination when stored on the floor in standing water. In the blower-washer. It presents a risk of cross-contamination from the non-molluscan animals to the bivalves. 3. or subjected to splash from foot traffic. Due to the nature of the shucking operation.3. the knife is used to cut the remaining adductor thus freeing the shellfish (meats). Thus. 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). through which the washed meats are discharged for rinsing and then packed in containers. Meats are collected in either perforated or non-perforated buckets for delivery to the washing station.2).2 Effects of microflora during processing The same considerations as previously discussed. If shellfish are not reasonably clean at the time of processing. a considerable quantity of the adhering material will be mixed into the shellfish during the shucking process. for the effects of microflora during the processing of shellstock. clothing becomes very soiled. are applied to the shucking operation of shellfish.2 Shucked shellfish 3.2. In addition. Another potential risk with the use of wet storage is the commingling of bivalve molluscs with other marine species such as crab. and fish. Shuckers routinely use either cotton or rubber gloves during the shucking operation. washing. Washing of meats with potable water is accomplished by placing the meats on a skimmer table (the stand-supported.1. To remove the shellfish attached to the remaining valve. the tank is opened through a drain gate and chute. whereas cotton become wet and 63 . the meats are in a tank of water and agitated by the introduction of air from the bottom of the tank. 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. 3. Containers are placed in packing boxes for shipping at refrigerated temperatures. Sealed containers are placed in boxes and covered with crushed ice for chilling or refrigeration and transportation. the finished product may become contaminated. Studies have shown the use of rubber gloves reduces the contamination level.3. removal of shellfish (shucking). After washing of meats. Under these conditions if the shuckers enter the packing room.1.3. thus contributing to high bacterial counts in the final product (28).1 Processing description The processing of shucked shellfish is limited to washing of shellstock.
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. 2. 64 . Shucking containers are rinsed clean with running water. Separate rooms or lockers for storage of clothing. There is a tendency to store such articles on the shucking benches or in packing rooms. 4. within one hour.FISH AND SEAFOOD retains bacteria during the shucking thus increasing the bacterial load in shellfish (29). and gloves help prevent the contamination of shellfish. Shucking buckets are completely emptied at the packing room and no overage is returned to the shucker. Shucked meats are thoroughly drained. which may lead to contamination of product. Shucking buckets and storage containers are kept at adequate height above the floor to prevent contamination from floor splash.2. aprons.2 ºC or less. cleaned and delivered to the packing room or prechilled and placed in temporary refrigeration (≤ 2 hours) at 7. Fig 3. and sanitised before each filling. 3.
The environment has an effect on the bacterial population found in shellfish. Much of the work has been done with oysters and clams. particularly oysters stored at 7 ºC. Flavobacterium.MOLLUSCAN SHELLFISH Unacceptable practices in the packing step can affect the microflora of shellfish. clean gloves. Shucked shellfish are an excellent medium for the growth of bacteria.8% (37). 3. shucking pails. For example.2 ºC or less. 31. 65 . and that storage in wet ice is the most effective method for refrigeration of shucked meats (30. in oysters. and packing the shellfish in a timely manner. 32). Controls that should be applied include using clean packing equipment. blowers and other utensils that have cracked. Micrococcus. The initial flora found in shellfish is the major factor of spoilage in bivalves. Glycogen. This process changes the bacterial profile to microorganisms favouring low pH. rough. glycogen levels range from 0. keeping shellfish refrigerated is the most effective method of retarding deterioration and spoilage. As described previously. Lactobacilli seem to be the predominant microflora in shellfish. abundant gas production. 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. skimmers.2 ºC. Pseudomonas. All equipment coming in contact with the shellfish such as colanders. Shellfish. 36). Shucked shellfish are held and transported at temperatures of 7. streptococci. particularly shucked meats. where there is a breakdown of temperature control or prolonged storage of shellfish. 33. 34. The spoilage process in shellfish may be divided into three stages: increase of acidity. thus producing acid and gas (38). In summary. anywhere during the process of shellfish from harvesting to the point of sale. Glycogen is hydrolysed by the initial flora found in shellfish.4 Spoilage There is a lack of information in the literature regarding spoilage microorganisms in shellfish.47 to 6. and Pseudomonas fluorescens are the responsible agents of spoilage in oysters held at various temperatures (10. spoilage will occur. at the turn of the century. these include lactobacilli. whereas bacterium such as Pseudomonas. and proteolysis (10). are an excellent medium for growth of bacteria. Studies have shown that bacterial growth is significantly reduced at storage temperatures of less than 7. Earlier studies. a carbohydrate. showed that organisms such as Achromobacter. 35. Refrigeration and shipping of shucked shellfish in wet ice is highly recommended. or inaccessible surfaces are apt to harbour accumulations of organic material in which bacteria may grow. Temperature abuse will result in high bacterial counts. Achromobacter and Flavobacterium levels decrease during storage. Alcaligenes. is stored in the bivalve as a source of energy. and yeasts. These organisms may play a significant role in the spoilage processes. Proteus.
Fortunately. although non-hemolytic strains have been associated with illness (43.1 Vibrio parahaemolyticus V.5. 3. The incidence of V. during processing. headache and fever. Large numbers of bacteria must be present to reach potentially infective levels of >105 colony forming units per g (39). parahaemolyticus from shellfish is low. V. shellstock and packing of shellfish in jars) (29). from harvesting to finished product (i.FISH AND SEAFOOD 3. and estimates based from a hospital study show less than 0. Descriptions of outbreaks or reports of growth of the organism in shellfish are presented. 66 . One of 250 cases reported involved shucked products with low levels. there were over 250 cases of illness on the West Coast of North American extending from California to Canada. Major outbreaks are associated with the warmer months.1. parahaemolyticus possibly producing illnesses (40). These organisms can be present in the shellfish from the point of harvesting.e. primarily in brackish or saltwater.5 Pathogens: Growth and Survival This section deals with bacterial pathogens frequently present in shellfish. 41). Products involved were from the same processor. nausea. parahaemolyticus causes gastroenteritis lasting 24 .5 cases/100. The ability of the bacterium to cause gastroenteritis is highly correlated with the production of a heat-stable haemolysin (42).000 population/year (46). 44). Diseases caused by Vibrios are enteric in nature. prevention of cross-contamination. only a small percentage of strains in the marine environment are potentially pathogenic (45). and harvesting of shellfish in the cooler months. where outbreaks occur regularly. coastal areas). there is a recent report of low levels of V. of V. and to the point of sale.1 Vibrios Vibrios are Gram-negative facultative rods naturally occurring in the marine water (estuaries. They can be introduced to the shellfish from the environment or added during processing.48 hours with abdominal pain. parahaemolyticus illness is usually a result of eating raw or partial cooked shellfish (8. This bacterium is present in all the major processing steps of a shellfish plant. The majority of the illnesses were associated with the consumption of raw or partially cooked oysters and clams. 3.5. Sporadic cases are frequent along the coast of the United States. however. V. lot and retail outlet. approximately 23 cfu/g. parahaemolyticus multiplies rapidly at temperatures above 20 ºC. It is a common marine bacterium. The control of growth and survival of these organisms can be reduced by the use of good manufacturing practices including immediate and proper refrigeration. and very common in Japan. parahaemolyticus. Between May and September 1997. ranging from epidemic cholera to sporadic cases of diarrhoea. diarrhoea. widespread in both polluted and unpolluted waters.
5. It is one of the most severe foodborne infectious diseases. 48). 58). abdominal cramps. those who have liver disease. lasting 6 . 57. 56). and preventing cross contamination once the product is cooked. Healthy individuals are susceptible to gastroenteritis. Death has been reported to occur within 36 hours from onset. 46.0. 54. and nausea within 24 . which have been implicated in shellfish-related disease outbreaks (31). cirrhosis.7 days. nausea. Onset of symptoms occurs in 48 hours and can be severe. Again. V. and rapidly become inactive at low storage temperatures.3 Vibrio cholerae non-01 Illness caused by V. Primarily.MOLLUSCAN SHELLFISH Control of V.1. vulnificus. primarily in immunocompromised individuals with a mortality rate exceeding 50% (50). cholerae non-01 is a common bacterial cause of molluscan shellfishassociated illness. in the range of 0. 46. vulnificus are fever. Control of V. 23 of 24 major USA-West Coast estuaries were positive for V. cholerae non-01 is generally a less severe gastroenteritis than that caused by V. They are sensitive to cold. 49).5. or primary septicaemia. The widespread distribution of this bacterium is comparable to other studies in the Gulf and Atlantic Coasts (52. V. cholerae non-01 is ubiquitous in the estuarine environment. 55. these individuals are advised not to eat raw shellfish. or immunosuppression. It causes diarrhoea. however. High-risk individuals. the organism has been found in the Gulf of Mexico. 53. In one study.2 Vibrio vulnificus The symptoms of V. parahaemolyticus growth in shellfish meats is temperature dependent.4 . V. gastroenteritis. Illnesses are usually associated with the consumption of raw oysters as described in Table 3. chills. The occurrence of V.8 case/100. All isolates are considered pathogenic. 3. Oysters are the most hazardous food associated with V. diabetes. 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 (8.48 hours of onset. The bacterium is widespread in estuarine waters particularly in warm waters of >20 °C. cholerae non-01 has been associated with septicaemia. parahaemolyticus the estimated annual incidence rate is very low. Hazards from this bacterium can be controlled by thorough cooking of shellfish. with a fatality rate of 50% for individuals with septicaemia. Control of oyster-associated 67 . cholerae. and vomiting. leukaemia. 3. Inadequate refrigeration will accelerate the growth of V. like V.000 population/year (8.1. The bacterium is an estuarine species that causes wound infections. the organism has also been found in cooler temperatures from estuaries of Washington and Oregon (47). fever. parahaemolyticus.II (8. cholerae non-01 (51). are particularly susceptible to primary septicaemia. vulnificus is by rapid refrigeration during warm weather months.
dehydration. Onset of disease is 6 hours to 5 days. They are common marine bacteria.FISH AND SEAFOOD V. cholerae 01 is transmitted by faecal contamination of food or water. There is evidence of strains of V.10% carbon dioxide for optimal growth conditions. Death may occur after severe fluid and electrolyte loss. widespread in both polluted and unpolluted waters. whereas excellent sanitation facilities in developed countries are responsible for the near eradication of epidemic cholera. 68 . the pathogenic severity is lower than V. 3.5. drying. 3.2 Campylobacter jejuni C. imported product from a country experiencing an epidemic.5. fluvialis. In general. processors should know the product source.48 hours with abdominal pain. and the transmission of the disease by consumption of raw. parahaemolyticus and V. hollisae. and harvesting of oysters in colder months when Vibrio counts in water are the lowest (59). nausea. and V. 60).5%) and 2 . Serotype Ogawa biotype El Tor and 0139 are responsible for epidemics in South American. In developing countries. nausea. 63).4 Vibrio cholerae 01 and 0139 Symptoms of epidemic cholera vary from mild watery diarrhoea to acute diarrhoea with characteristic rice water stools. and illness lasts 7 .S. including V. There are several other Vibrio spp. however. 62. vulnificus. and shock. low pH. The presence of these strains appears to be in low numbers and non-virulent (59). In addition. or cross-contaminated shellfish (57. Illness can include abdominal cramps.g. undercooked. jejuni requires reduced levels of oxygen (3 . and southern Asia including India and Bangladesh. The frequency of infection of these organisms is no different from other Vibrios. V. preventing crosscontamination of cooked seafood. particularly in oysters. diarrhoea. is the same as previously discussed for the non-01 strains. Gulf coast environment. (8.1.1. 3. It is considered a fragile and sensitive organism to environmental stresses such as oxygen.5. acidity and heat. V. Poor sanitation and contaminated water supplies will spread the disease. fever. these organisms cause gastroenteritis lasting 24 . respectively. The control of V. cholerae non-01 illnesses is by cooking seafood thoroughly. salinity. cholerae 01 establishing in the U. Onset of illness is from 3 to 5 days after ingestion of contaminated food. mimicus. abdominal pain. cholerae 01 in molluscan shellfish. Control of infection from these organisms is no different from the other Vibrios.10 days. associated with shellfish-borne illness outbreaks.5 Other Vibrios spp. It causes diarrhoea. V. Illnesses are associated with the warmer months. and nausea. vomiting. headache and fever. cholerae 01. e. 61.
5. V. Although. V. 73). the organism is being continually inoculated into the environment by waterfowl and farm run-offs and thus eventually contaminating shellfish growing waters. control of C. 78) and P. ranging from 0. 76. 81. In one case. Proper care should be taken if the finished product is to be packaged in modified-atmosphere packaging (MAP). The seasonality of 69 . and have been implicated as the causative agent in illnesses from the consumption of raw oysters and clams (68. jejuni does not multiply in shellstock at refrigerated temperatures. hydrophila group (75. Campylobacter spp. infection occurred among participants of a firemen's banquet. 80) are ubiquitous in the marine environment. Campylobacter appear to survive well in oysters during storage. parahaemolyticus. from Pacific shellstock oysters purchased from a retail grocery store (74). however. jejuni were recovered. To reduce the risk of shellfish-borne campylobacteriosis. however. 77. cholerae 01 and non-01. 84. 82. 65. V. fluviallis. jejuni survived for over 20 days in market-shucked oysters stored at 4 ºC. mimicus. 3. and V. although their role as a pathogen is highly controversial and circumstantial.MOLLUSCAN SHELLFISH TABLE 3. This finding is significant in that C. V. Recently. 66). whereas at 10 ºC Campylobacter survive only for 5 days (71). of Cases 4 362 0 1 0 a Includes V. 83. have been isolated from shellfish beds throughout the world (67.II Molluscan shellfish associated outbreaks caused by Vibrio spp. They have been implicated in limited shellfishassociated gastroenteritis cases (8.3.3 Other organisms of concern 3.500 cells/gram of Campylobacter may cause illness (64. hollisae Infective dose varies with host susceptibility and virulence of the strain. 69. Sixteen of 28 participants became ill with campylobacteriosis after ingesting raw clams. they survive well at refrigerated temperatures. 71).1 Aeromonas hydrophila group and Plesiomonas shigelloides A. shigelloides (79.4 to 114 cells. 72. and their immediate proper refrigeration. Human feeding studies suggest that about 400 . the incidence of Campylobacter infection is low from the consumption of molluscan shellfish. jejuni begins in the harvest of shellstock from approved waters.5. C. Campylobacters do not survive well in the environment due to the organism's lack of resistance to environmental factors. 70. of Incidents 4 279 0 1 0 No. 68. vulnificus.a (8) Species Hard clams Oysters Soft Clams Mussels Scallops No. 85). C.
They are easily destroyed with mild cooking temperatures. FDA/Administrative Guidelines 3 Source (91). coli Salmonella V. after 7 days storage of shucked oysters at a shellfish processors’ cooler kept a 5 ºC Aeromonas counts ranged from 500. and where indicated are intended to be used for official control purposes. parahaemolyticus Staph. 2 Source (90). In one study. FDA/ Administrative Guidelines.000 cfu/g1 <230/100 g2 <1000 per gram. 70 . Enforcement action is considered on a case-by-case basis taking into account all the factors associated with the specific situation. jejuni V. dried milk/whey drink powders.000 MPN per gram1 negative for staphylococcal enterotoxin or when the viable MPN count is <10.FISH AND SEAFOOD Aeromonas and Plesiomonas is similar to the Vibrios. suggesting that these individuals are at higher risk when consuming raw or partially cooked shellfish containing these bacteria. vulnificus 1 Acceptable Levels in Molluscan Shellfish <500. coli Enterotoxigenic E. Control of these organisms in shellfish products is limited. 87) of the outgrowth of A. being isolated more often during the warmer month. 4 FDA and ISSC recommending establishment of an educational program to alert at risk population of presence of V. Caution should be exercised by individuals with haematological malignancies (88).000 to 3. soy products. prepared sandwiches.600. aureus C. 3.0001 negative for the presence3 Under review4 Source (89). Note: Although molluscan shellfish are not included in the selected high risk food. Test Standard Plate Counts E. cholerae V. Similar results were seen by other investigators (86. vulnificus in molluscan shellfish. action is considered on a case-by-case basis taking into account all the factors associated with the specific situation. non-dairy frozen desserts. Pathogen monitoring of Selected High Risk Foods Domestic including: milk. hydrophila group stored at refrigeration temperatures. FDA established action levels or levels of concern for certain microbial pathogens in molluscan shellfish. The Agency will consider enforcement action against the shipment of molluscan shellfish if the levels exceed as stated. They survive well in shellstock or shucked oyster meats held at refrigeration temperatures.000/gram (29).6 Published Microbiological Criteria The published microbiological criteria below are included as a guide. 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.
544-8. 447-52. Gill S. 1926. 10.K. Bacterial flora of the market oyster. 1994. Ed. water and sediment from Galveston Bay.M.K.R. 8. 44. Jr. Smith F.E. 1936. Organisation for Economic Co-operation and Development.S. 105-8. Microbial flora and level of Vibrio parahaemolyticus of oysters (Crassostrea virginica).7 1. in Microbiology of Marine Food Products.R. Liston J. 64. 1982. Applied and Environmental Microbiology. 1970. Cook D. American Journal of Hygiene.W. Ward D. 3. Microbiological characteristics of Dungeness crab (Cancer magister). Hackney C.H. 72-8. Occurrence of the spirochaete genus Cristispira in western Canadian marine bivalves. 13..S. The American Oyster. 71-6. Microbiology of shellfish: bacteriological study of the natural flora of pacific oysters (Crassostrea gigas). Wiley-Blackwell.J. 1975. Colwell R. 26. 30. Rippey S.E. in General Zoology. 7. 33643. 104-9.B. Biology of the oyster in relation to sanitation. Journal of Milk and Food Technology. 58. Thompson C. Applied Microbiology. 42. Microbiology of bivalve molluscan shellfish. A typhoid fever epidemic caused by carrier bootlegging oysters. 71 .S.. 1960. in Fishery Bulletin 1964 No. Walker W. USA. Villee C.C.R.L.C. 1964. Philadelphia.. Colwell R.. 12.R. Eds. Schmidt C. 15. 1968.MOLLUSCAN SHELLFISH 3. 1991. Pfeifer D. Viability and release Salmonella charity and Escherichia coli from oyster faeces..W. Galtsoff P.. 11.R. Nauman R. Ray S. Jr.D. 1940. Washington D. U. Proceedings of the National Shellfisheries Association. Rowse A. Eds. Tubiash H. 1981. 19-34. Cook D. Saunders Company. Applied and Environmental Microbiology. New York. 9. Tall B.A. 14... Clinical Microbiology Review. Fleet G. 30. Eliot C. Galtsoff P. 755-76. 46. 1984. 633-40. 1995. 82-7.. 17. 13. Applied Microbiology. 6.S. 7. Infectious diseases associated with molluscan shellfish consumption. Hood M. Vanderzant C. Lee J. American Journal of Public Health.. 1973. 16. 36. Lovelace T. Effects of harvesting waters and storage conditions on yeast population in shellfish. Fish and Wildlife Service.. 29. References The Phylum Mollusca. 231-51.F. 2. Journal of Food Protection. 1983.N. 6. 8. Veliger.. Journal of the Mississippi Academy of Sciences.R.A. Van Nostrand Reinhold. Quantitative and qualitative commensal bacterial flora of Crassostrea virginica in Chesapeake Bay. 1963. W. American Journal of Public Health. 245-7. in Multilingual Dictionary of Fish and Fish Products. 33-6. Fate of enteric bacteria in estuarine sediments and oyster faeces. 4. 5.. 419-25. Old H. Scanning electron microscopy of Cristispira species in Chesapeake Bay oysters. Bernard F. Government Printing Office.A.
H.. Ed.J. Jones W. Kelly C. 123-8. 47 (5).S. 989-1003. The influence of time and temperature on the bacteriological quality of shell oysters during processing and shipping..T. 1971.S. Influence of U.W.. 1995. Fraiser M. Fujino T. Indicator bacteria and Vibrionaceae multiplication in postharvest shellstock oysters. 31. Sakazaki R. Crassostrea virginica and Mercenaria campechiensis..A.. 24. Hepatitis epidemic conveyed by oysters. 21.A. 72 . 1967. 29. Abeyta C. F61-9. 1956. Applied and Environmental Microbiology.. Handling and Shipping Shellfish.E.J. 1205-10. I.FISH AND SEAFOOD 18. 25. 30. 1964. 50.. 1989. Stott R. Kaysner C.S. U. 69. 1980.D.. Son N. Trost P. Beck W. Bacteriological studies of commercial shellfish operations on the Gulf Coast..B. 53. 1985. Behaviour of pathogenic bacteria in the oyster. Cook D. Establishment of critical control points in west coast oyster processing plants. 4. Journal of Food Science. U. Ness G. during depuration. Temporal occurrence of Vibrio species and Aeromonas hydrophila in estuarine sediments. 5.F. Bacteriological survey of an oyster bed in Tangier Sound. Presnell M. and storage.. Public Health Service Technical Report.W. Applied and Environmental Microbiology. Applied and Environmental Microbiology. Sadaguchi G.A. Part II Harvesting. Food and Drug Administration. Maryland. 1221-8.. Liston J. Chesapeake Science. 1214. 27. re-laying.. 1973. 994-1002. Crassostrea commercialis. Bacteriological changes in live shellfish: Pacific oysters (Crassostrea gigas). in International Symposium Vibrio parahaemolyticus. Vasconcelos G. 40.J. Arcisz W. 1961.. 22. Rodrick G. seafood-handling procedures on Vibrio parahaemolyticus. Food and Drug Administration.C. Eds. Svenska Lakartodningen. Ltd. Wekell M. Timetemperature effects on the bacteriological quality of stored shellfish. and Manillal clams (Venerupis japonica). 1992. Kelly C... Fleet G. 23... Survival of enteric organisms in shellfish. U. Journal of Food Protection. oysters.A...S.W. crabs and mullet..H. 45.W. Jr. Journal of Food Protection.B.E. P-1..A. 32.S. Public Health Service. Williams L. 343-9. Special Report U..M. Roos B. 1954. LaRock P. 1490-5. 1984. 20. Hoff J. Food and Drug Administration. 26. Koburger J. Vaughn M. 52. Saikon Publishing Co. Ruple A. Blake N. Native Littleneck clams (Protothaca staminea).A. 343-5. 1983. 167-71. Ericksen T. Takeda Y. FDA Gulf Coast Technical Services Unit. American Society for Microbiology. 324.B. Effects of storage on microbial loads of two commercially important shellfish species. in National Shellfish Sanitation Program Manual of Operations. Tokyo.G. Olympia oysters (Ostrea lurida). Presnell M.W.C. Hood M. Public Health Report. 19. 28. Incidence of Samonellae in clams. Colburn K. Washington D.
273-80. New York.B. Vibrio parahaemolyticus illnesses. Crassostrea virginica. The bacterial content of market oysters. 41.. Infectionskrankheiten und Hygien. Journal of Food Protection. 42. 43. Takikawa I.. Shellfish Specialist. 1987. Hlady W. Sample T. 160.. 1916. 313-22.M. Kaysner C. 1969. Applied Environmental Microbiology. Volume 1. Berry F.G. 1954.A. 107-8.A. 1958.D. Leitch J. Stroh E. Sidwell V. Bothell.S. Kelly M..W. 1976.. 33. kanagawa-negative Vibrio parahaemolyticus from patients and the environment in the Pacific Northwest. 41. Takizawa K.. 1937. 41. Bacteriological control of oysters during processing and marketing. Akiyama S. Ed. Fieger E.A. 1989. Kato T.D.C. Miyamoto Y. 13-7. Ishibashi M. Journal of Food Science.. Washington Report October 14... Novak A. 47. 69.. 100. Bulletin of the Johns Hopkins Hospital.. Microbiology of shellfish decomposition.W. 46. 97. Takeda Y. 53.. 38.M. Israel E. 2820-2. Serological and hemolytic characteristics of Vibrio parahaemolyticus from marine sources. The bacterial flora of market oysters. 1926. Yokohama Medical Bulletin.. Hoge C. Grodner R. Studies on pathogenic halophilic bacteria. Epidemiology and spectrum of Vibrio infections in a Chesapeake Bay community. Geographic and monthly variation in composition of oysters. Public Health Reports..M. Journal of Clinical Microbiology. Obara Y. 39. Virulent strains of Vibrio vulnificus isolated from estuaries of the United States west coast. Morris J. Journal of Clinical Microbiology.P. 60. Urease-positive. 45.T. 1914.. 27. 1.. West Coast States and British Columbia. Zentralblatt für Bakteriologie. 37. Vanderzant C. 73 . 40. Emergence of a restricted bioserovar of Vibrio parahaemolyticus as the predominant cause of vibrio-associated gastroenteritis on the West Coast of the United States and Mexico. Peeler R.. Food and Drug Administration. Jacobson M. Libonati J. Marine Fisheries Review. 27.MOLLUSCAN SHELLFISH 32. Watsky D.. 44...L. 1349-51... Journal of Infectious Disease.. Powers C. 25.. Jr. 1989. Bacteriological examination of oysters stored at low temperatures.. Journal of Bacteriology. Borgstrom G. 1147-9. Parasitenkunde.S. Academic Press. in Fish as Food. 133-47.A. Journal of Infectious Disease. Abeyta C. Kelly C. Stott R.N. 1979. 1961. 2891-3. 353-7. Arcisz W. DePaola A. Kaysner C. 36.F. Joseph S.E. U.. Vibrio infections associated with raw oyster consumption in Florida. Jr. In vitro haemolytic characteristics of Vibrio parahaemolyticus: Its close correlation with human pathogenicity. 985-93. 2. Geiger J. 34. Janda J. Yamai S.L. Thompson C. U. Bacteriological findings in Baltimore oysters. Loomis A. 1989.M. Wekell M. 1997. 204-5. (2)..G.. 35. 1997. 128-31. Ward W.. Tanikawa E.. Joseph M. Journal of Bacteriology.M. 38. Abbott S.
.... Hug A.A. Jr. Jr.J. Blake P.A. 60. Joseph S..M. Wazenski T. Kaysner C.T. Journal of the American Medical Association.. 53.M. DePaola A.A.. 91-103.J. Lancet. 1979. Stott R.W... 1980. 56. Presnell M. Spite G... Colwell R. Feldman R. Zywno S. New York. 37.A..T.A. McFarland L.O. Francis D.M. Roderick G. Kaper J.. cholerae in Louisiana.W. Characterisation of Vibrios cholerae isolated from oysters. 1982. 74 . Pons V. 8.E. Morris J. 94. 305-9. Hollis D.. 45. John Wiley & Sons Inc. Craig J. 1983. 1984. Journal of Food Protection.R. 1985.. Gulf Coast.. Jr. 1988. Vibrios in the Louisiana Gulf Coast environment.T... 46.. Jacobs R. Snyder J. 10. 57. Applied and Environmental Microbiology. Lockman H.C.J..V. 1012-7. Davis B.. Clinical features and an epidemiological study of Vibrio vulnificus infections.F. Tacket C..M..M. Wathen H.E.D.G.G. Brenner F. Non-01 Vibrio cholerae in shellfish.. Morris J. Small P.O.. Blake P.T. Abeyta C. Illness caused by Vibrio damsela and Vibrio hollisae. Twedt R.. Duran W... Non-O group 1 Vibrio cholerae gastroenteritis in the United States: Clinical.G.W. McFarland L. 49. Becker S.. Allegra D. 1983. 656-8. Johnston J. Jr.G. 275-83. Hug M. Peeler J.R.. Wachsmuth I. and enterotoxin production of Vibrio cholerae in Chesapeake Bay. Vibrio vulnificus: Man and the sea.. Wilson R. 52.A possible endemic focus in the United States. West P. New England Journal of Medicine.. Siebeling.A. McPhearson R... 53..P. Plaard R. Conte J.F. Riddle C.G.. 41. Bradford H. 51. New England Journal of Medicine. Bradford H. DePaola A. serology.. Applied and Environmental Microbiology.M.F.A.B. 1344-8. 61..M..T. 253. Annals of Internal Medicine. 1.S.. 343-50. McCay C. 1987.B.G. Lee J. 55. 305. Incidence of Vibrio cholerae from estuaries of the United States West coast. in Vibrios in the Environment.M..FISH AND SEAFOOD 48..N. Blake P. Ecological relationship between Vibrio cholerae and planktonic crustacean copepods. Ecology. Microbial Ecology.. 1475-8...M. Colwell R.A. sediment and waters of the U. 1294-7. Twedt R.. Black R. Applied and Environmental Microbiology. 312.. Wekell M. 50. Roberts N. Wilson T. 1985. Cholera and other vibrioses in the United States.C.L. Journal of Infectious Disease. Hebert S.. Jr. Blake P.R.. Miller H. Motes M. Ed. Jr.G. 58.. 558-61.B..E.W. Applied and Environmental Microbiology.E. 1984. R. 54. Puhr N. An epidemiological study of V. Non-01 Vibrio cholerae bacteraemia: Case report and review. Feeley J. Rahman R.A. Leitch J.G.. and laboratory characteristics of sporadic cases. Morris J. Adams M.. 5972. 299-312. Safrin S. Maden J. Barrett T. Jr. Becker R. 1982.. 1981. Review of Infectious Disease. 2050-3.. 59.. Morris J. 802-6.I.. Colwell R.R. Hunt J. Cholera ..O. 1981.D... Weaver R... Kaper J... Tacket C. epidemiologic.K.. Caraway C. 149. Hickman F..
D. a newly recognised Vibrio species.L. Journal of Food Protection. 472-9.J..I.. Campylobacter jejuni in a Washington State shellfish growing bed associated with illness. 169-71.R.G.M. Huq M... Rowe B. Wilson T..G. Alam A.A.M. 6-9 September 1983. Infective dose of Campylobacter jejuni in milk.M. Plenum Press.A. Helicobacters and Related Organisms.W... Clements M.J. 65. Deeter F. 1993. Pearson A. 1991. Blaser M. Ireland.A. Feldman R. Murphy P. Kaysner. Journal of Rapid Methods and Automation in Microbiology. 607-9. Journal of Clinical Microbiology. Stott R. Incidence and survival of thermophilic campylobacters from shellfish growing water: media evaluation. Eds. 4.R. 56.M. Campylobacter gastroenteritis associated with raw clams. Brussels.A.F. Hunt J. C. Part 1 Abstracts: Shellfish as a possible source of thermophilic Campylobacter spp.P.. Robinson D. Levine M.J.. Matsuda M. Eds. 11.K.L.. S41. Wareing D. 4.X. 1980.G.. Morris G.. The occurrence of Campylobacter jejuni and Campylobacter coli in Sidney rock oyster.S. 282. Black R. 75 . 64. Skirrows M. 63. 71. Proudford R.M. Microbial Ecology in Health and Disease.A. Davis J. 73. London. Disease from infection with Vibrio mimicus. Murphy P.. Helicobacters and Related Organisms. Annals of Internal Medicine.. Isolation of vibrio-like group.E.G. 1987.S. 157. 1995. 68... infections in man. 4.. Kaysner C.. 323-5. 66.H.. New York. Newell D. New York. Jr. Jr. Griffin M. 1980.K.G. Wareing D.. Shandera W. Levine M. Wilson I. 80. Wilson T.. Arumugaswamy R. Public Health Laboratory Service. British Medical Journal. Jr. 1981. Experimental Campylobacter jejuni infection in humans. Newell D. Wilson I. Fitzpatrick M. 72. 1988.P. International Journal of Food Microbiology. Abeyta C..D. 257.. Black R. 1584. Hughes T.. Journal of Infectious Disease. 101-4.E.. A method for isolation of Campylobacter spp. Journal of the Medical Society of New Jersey. 70.. Kaysner C.. Plenum Press. 67. from patients with diarrhoea.A. 256.R. 13. coli and lari) from shellfish and the marine environment.. in Campylobacters.G. in Campylobacter II: Proceedings of the Second International Workshop on Campylobacter Infections. Johnston J. EF-6.R.. Eds..E. Clements M. Moore J.R.. Blaser M... Abeyta C.. (jejuni.A.M. Jones D. Phenotypic and genotypic characterisation of urease-positive thermophilic Campylobacter (uptc) isolated in N..M. Davis B.. Wekell M.... Blake P.K. 1983.B. Huges T. Studies of Campylobacter jejuni infection in volunteers.A.G.E. 51-64.M. 1983. 621-4. Moore J.MOLLUSCAN SHELLFISH 62.. Ketley J.J.. Feldman R. 99. Austin S. 69. Brenner D. Dalley E. in Campylobacters. Abeyta C. 1996.. 1996.A. Wekell M.M. Ketley J.
Herrington D. Fliermans C.. Incidence of motile aeromonads from United States west coast shellfish growing estuaries.J.. 1989. Wekell M. Van Damme L. Jr. Kaper J. Holmerg S. 1984. 1146-9. Vandepitte J. Partridge K.A. 55. Abeyta C. vibrios and Aeromonas hydrophila. Levine M. Department of Health and Human Services/Public Health Service/Food and Drug Administration. 449-57.R.. Wachsmuth I. 690-4. The use of bacterial membrane fractions for the detection of Campylobacter species in shellfish. Plesiomonas shigelloides: An opportunistic food and waterborne pathogen.. Effects of processing and storing oyster meats on concentrations of indicator bacteria. 76. Journal of Food Protection..W. 1978. 998-1001. Wekell M. Farmer J. Hickman-Brenner F.. Kaysner C. Recovery of Aeromonas hydrophila from oysters implicated in an outbreak of foodborne illness. 7-12... Cook D..FISH AND SEAFOOD 74. Aeromonas hydrophila: ecology and toxigenicity of isolates from an estuary. Stott R.P.C. 731-8.. In vitro and in vivo pathogenicity of Plesiomonas shigelloides. epidemiology. 82. and treatment of Plesiomonas shigelloides diarrhoea. Petersburg. Atlanta. Fla. 80. 1980.. Infection and Immunity. Baker R. 86.. 1987.M.. Indicator bacteria and Vibrionaceae multiplication in postharvest shellstock oysters.M. 1988. Herrington T.K. 81.A. Kain K. 1997.. 48.. Hirsch R.M. 105..A.A. Kaysner C. Journal of Rapid Methods and Automation in Microbiology. 1989. Jr.. Prevalence and distribution of Aeromonas hydrophila in the United States. Koburger J..W.M. Journal of Food Protection.. 27. 79. Kelly M. Journal of Applied Bacteriology.M. Robins-Browne R. Bark D. Tzipori S.J. 84.D. 85. Tall B..A. 5... Peterson D. Journal of Clinical Microbiology. 849-55. 1990. Wekell M. Burke V.. 1985. Journal of Food Protection. 75.. Beaman J... Applied and Environmental Microbiology. Lockman H.. 223-47. 979-85.M. Gracey M.J.. 87. Hazan T. Applied and Environmental Microbiology. 47.B. 39. Journal of Food Protection. 50. Journal of Food Protection.L. 52. 598-601. Hunt J. Trost P. Jr.T. 83. Hood M. Seidler R. Report (1/13/84). Biotyping and virulence factors in clinical and environmental isolates of Aeromonas species.N.F.L.A. Miller M..R. 78. Clinical features. 343-9.. Kaysner C. 1984... 475-9. Colwell R. GA. Frequent isolation of Edwardsiella tarda and Plesiomonas shigelloides from healthy Zairese freshwater fish: A possible source of sporadic diarrhoea in the tropics.. 52. 77. Plesiomonas enteric infections in the United States.C..M.. Abeyta C. Sullivan J. Cooper M. 53..L. Ruple A.D. Singelton F. 76 . 36. Tenge B.D... 1986..H..A. Robinson J. 1981. III.J. 359-77. Blake P. Stelma G.A. Esch G. Abeyta C. Annals of Internal Medicine. 47. Oyster borne disease outbreaks in St. Applied and Environmental Microbiology.W.
Food and Drug Administration. Burdge D.1987. and HACCP: A Global Focus.S. Slavin J. Government Printing Office .R. Pittsurgh. National Academy Press. Program 7303. U. 63-8. 3.L.S. U..E. Seafood Safety.S. Martin R.W. Sherlock C. Ed. 560.Pathogen monitoring of selected high risk foods (FY 88/89).C. mussels. Clams. Gaithersburg. Fish Inspection. Chapter 5: Foods. Lund B. Food and Drug Administration Compliance Policy Guides.A. Washington D. in U. 90.600 (CPG 7119.MOLLUSCAN SHELLFISH 88.. U. 7. 91.C. 77 . Washington D.12).W. Smith J. Ed.H. U. Technomic Publishing Co. Ed. Quality Control.C. Food and Drug Administration Compliance Program Guidance Manual.. in U. in National Shellfish Sanitation Program Manual of Operations. Food and Drug Administration..S. 89. Washington D. Aspen Publishers Inc. 1997.8 Further Reading Ahmed F. Food and Drug Administration.E. Part I Sanitation of Shellfish Growing Areas.M. Food and Drug Administration. 2000. Food and Drug Administration.. Baird-Parker T..S. Does Aeromonas hydrophila preferentially colonise the bowels of patients with hematologic malignancies? Diagnostic Microbiology and Infectious Disease. The Microbiological Safety and Quality of Food. Food and Drug Administration. 1991. Collette R. Lancaster..S.S. Food and Drug Administration. Inc.C. fresh or frozenadulteration by bacteriological contamination Sec. 1995.S.. oysters. Food and Drug Administration.030. 1987.. Gould G. 1995. U. U.
packed and frozen in 0. e. and are usually aquatic. Shrimps. and frozen in 1 and 2. and available in 0. lobsters and crabs have ten legs. jointed limbs and a chitinous exoskeleton.5 and 1.0 kg polythene bags.1 Definitions Crustaceans are invertebrate animals and represent a class of the phylum Arthropoda.5 and 1. Shrimp can be purchased in the raw or cooked state in a variety of forms: Entire shrimp. M. Peeled ready-to-cook shrimp are raw. Peeled and cooked shrimp are ready-to-eat after controlled thawing. 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. peeled product. FRSPH Ethical Trade and Food Management Group Natural Resources Institute University of Greenwich Central Avenue Chatham Maritime Kent ME4 4TB United Kingdom 4. prawns and scampi are temperate species commonly eaten by man and include members of the families Nephrops norvegicus (scampi).5 kilo pack (boxes or bags). (deep-water shrimp). They possess a segmented body.4. Pandalus borealis (deep-water shrimp). 79 ... the front pair ends in claws..Ph. crabs. lobsters. crayfish and shrimp (1). Crayfish. Pandalus montagui and Crangon crangon (inshore shrimp). Parapeneopsis spp. CRUSTACEAN SHELLFISH Linda Nicolaides. Such products are usually only partially cooked before freezing. and Trachypenaeus spp. with additional water to form a glaze.0 kg polythene bags. Species eaten from tropical waters include Penaeus spp.g.
lobsters and crayfish are traditionally kept alive after capture and then cooked to kill immediately before consumption. If these products are subjected to temperature abuse. Following capture or harvest. Poor hygienic procedures will increase the probability of contamination with bacteria of public health significance.. for crabs and lobsters. particularly after it has been cooked. the level and type of the initial microflora of crustacean shellfish will reflect a combination of factors. during the peeling of cooked shrimp. Pasteurised crabmeat is edible meat removed from the exoskeleton by hand. Shewanella putrefaciens and 80 . e. shrimp die immediately after capture. which means that the haemolymph can be a reservoir of bacteria. Spoilage begins following death once the immuno-response system of the crustacean shellfish fails to function. their feeding and living habits. traditionally. hence. However. stored and transported as frozen product. The microbial flora of crustaceans from temperate waters differs from that of crustaceans from warmer. Control measures to prevent these potential hazards occurring include training and supervision of operatives as well as Good Hygienic Practices (GHP). only live carcasses are processed. Microbiological risks associated with crustacean shellfish increase in proportion to the degree of handling that the product undergoes. Spoilage bacteria associated with temperate crustaceans include Pseudomonas spp. spoilage bacteria are of little importance as. In comparison. the geography of the area they are captured from or are grown in. or frozen crab. it is important that shrimp are handled and iced correctly in order to keep them in as fresh and uncontaminated a state as possible. As with finfish.FISH AND SEAFOOD Crabs. particularly for members of the genus Vibrio (2).g. so that spoilage may occur during landing and transportation to the processing plant. boiled to kill the crustaceans and cook their meat prior to consumption. the potential exists for crosscontamination from both the processor and from the environment. e. Cooked and chilled. 4. The structure of the circulatory system of crabs is not closed. which include the environment from which they have been captured or harvested. tropical waters. this presents more favourable conditions for growth by both spoilage and pathogenic bacteria. lobster and crayfish are available for the consumer owing to the growing demand for ready-to-eat products. storage above the recommended storage temperature.2 Initial Microflora As with finfish. packed into polythene bags. the flora will change depending upon the methods of handling and/or the environmental conditions to which they are exposed. there is a growing market for fresh lobster tails or cooked crabmeat in crab shells. with the increasing demand for ready-to-eat/cook crustacean products. and the temperature and quality of the waters in which they exist. lobster tails and cooked crabmeat presented decoratively in their shells.g. the season. For example.
thus.for example.. Shrimp might be further processed with the shell on. A large proportion of shrimps are produced by aquaculture. Alcaligenes. by removing the head and thorax a source of enzymes is eliminated that will prevent the occurrence of black spots. lobsters and crayfish are harvested from cages or pots containing bait. especially if they are stored in ice or iced water. or are found in brackish water around the shores. When the shrimp arrive at the processing plant they are washed and then sorted into size. from the time of capture and processing. peeled and cooked. After capture and transport to the processing plant.3. or with the shell and central vein removed. uncooked prawns and shrimp is presented in Figure 4. or cooked. so they are either sold on the local market or are processed in local processing plants. 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. In contrast.1. graded according to size. Vibrio spp. These animals can survive out of water if the surrounding environment is kept cool and damp. 4. Shrimps can be processed whole.3 Processing and its Effects on the Microflora Crabs. and will be dependent upon whether the crustaceans were harvested from cool or warmer waters. the rate of deterioration of the crustacean shellfish by these organisms is reduced. chilled sea water.CRUSTACEAN SHELLFISH members of the Acinetobacter-Moraxella group. Pseudomonas spp.. Scrupulous hygiene is important at this stage to prevent the cross-contamination of the shrimp tails. or transported directly to processing plants. whether GHP are in place. In comparison shrimps die quickly after capture. 4. the genera associated with tropical species include Enterobacteriaceae. Hence. with “head on” or “head off” whereby the edible tail is removed from the head. they are washed. As the muscles of crustacean shellfish decompose rapidly after death dead animals are rejected.1 Prawns and shrimps An example of the process used for preparing frozen. The rate of spoilage will be a function of time and temperature.1).10 days for temperate water species (3). The latter process can expose the shrimp muscle to incidents of cross-contamination if GHP are not followed. chilled 81 . They are then taken to market alive in either holding tanks. glazed and frozen (see Figure 4. gills and thorax. Micrococcus spp. bacterial contamination and spoilage are inevitable. Shrimp will then be glazed and frozen in a raw state.. there is evidence that the shelf life of tropical crustaceans is longer than that of temperate-water species stored correctly in ice . up to 16 days for tropical species compared with 8 . The claws are usually bound together to prevent the animals fighting and causing damage to each other. As with finfish. and Achromobacter spp. however. corynebacteria.
1. particularly the pathogens Bacillus cereus and Clostridium perfringens originating in the flour.FISH AND SEAFOOD frozen. Good Manufacturing Practice (GMP). After cooking. 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. Breaded products introduce an added risk of spore forming bacteria. The cooking stage will eliminate the intrinsic microbial flora of the shrimp as well as any contaminants that have been added during handling and transportation to the processing plant. and packed to meet the customer’s specification.frozen uncooked prawns 82 . Fig. Flow diagram . 4. spices and cooking batter.
In comparison.2 Crabs and lobsters Crabs and lobsters should be alive when boiled or sold. If the tail drops. Heat is produced if the shrimps decompose in a confined space.3. slightly iodine smell. they include Sh. the tightness of limbs will indicate whether they were killed by immersion in boiling water or. 4. 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. whilst boiled crustaceans have a pleasant slightly sweet smell. If boiled. 83 . wet and ‘soapy’ or ‘jammy’ to the touch. the meat is either repacked and sold in the shells. 4.4. Processed crab and lobster meat can also be bought as packages of meat that have been subjected to a pasteurisation step. Once removed and cleaned. or packaged and processed. putrefaciens and Pseudomonas spp.2 Cooked crabs and lobsters As indicated earlier.CRUSTACEAN SHELLFISH 4. 4. or in hermatically sealed cans that have undergone an appropriate heat treatment. there is a new hazard of contamination of the cooked meat by workers during its physical removal from the exoskeleton. there is no associated spoilage problem. immediately prior to boiling. dull and sticky.4. contaminating microflora may include enteric bacteria. the joint between the tail and carapace is open and the limbs hang loosely. and have a sweetish. with visible decomposition or green discoloration seen through the membrane exposed by the dropping tail. viruses or parasites. If hygienic procedures are not followed. The shells have a crisp. putrefaciens. However. and a strong ammoniacal odour is produced. since crabs and lobsters are traditionally consumed immediately after cooking. An offensive odour is also produced. indicative of protein degradation and the growth of the principal spoilage bacterium Sh. bacteria of public health significance. The shell will be soft. With spoiled crabs and lobsters.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. spoiling prawns and shrimps have shells that are soft. for crabs. with the growing demand for ready-to-eat products from crab and lobster meat. there is no indication of life.
monocytogenes was absent from 40 samples of cooked prawns. They are mesophilic microorganisms usually residing in the intestinal tracts of human beings and warm-blooded animals. 4. 7). 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. monocytogenes in cooked ready-to-eat crustaceans is recognised as a potential risk to selected groups of individuals: the very young and 84 . Salmonella cannot be completely removed from raw shrimp during processing. in 24. shrimp and shellfish (11). monocytogenes in contaminated crustacea was recognised in the United States of America (USA) when the pathogen was isolated from Mexican frozen cooked crabmeat (10). and to increase testing of many other domestically produced seafoods under the General Pathogen Surveillance Programme (14.FISH AND SEAFOOD 4. animal manure. nearly 30 years elapsed before the potential hazard of L. The presence of L. The above results prompted the FDA to develop a compliance programme for the pathogen. later surveys in 1990 recovered the pathogen from retail raw. although efficient cooking will quickly eliminate it (6).5. An import alert was put in place in June 1987. One of the earliest accounts of the presence of Listeria monocytogenes in crustaceans was by Russian workers in 1959 (9). Results of a survey carried out in England and Wales between 1987 and 1989 demonstrated that L. and usually for both.5% of frozen raw shrimps from different countries (13). 16). Although foodborne illness had not been associated with the product. even if GMP is followed. often contains Salmonella and other enteric pathogens (8) and has been demonstrated to contaminate crustaceans growing in these environments. which required automatic detention of all Mexican frozen crabmeat that tested positive for Listeria and Escherichia coli (12). 6.5. as well as for Salmonella.5 4. Further surveys identified Listeria spp. 5. monocytogenes in cooked ready-to-eat foods. fish fingers. a common pond fertiliser used in aquaculture. the first in a series of Class I recalls was issued in May 1987 to retrieve nearly 4 tonnes of tainted crabmeat (11). Following this.cooked prior to consumption. Aquatic birds and the hands of workers have also been identified as sources of the pathogen. shrimps and cockles sold (17). cooked and smoked fish. However. in compliance with the Food and Drug Admistration’s (FDA) ‘zero tolerance policy’ for L. in domestic/imported shrimps (15).1 Pathogens: Growth and Survival Salmonella spp. Moreover. The genus Salmonella contains a wide variety of ‘species’ pathogenic for man or animals.2 Listeria spp. The pathogen was later identified in frozen raw shrimp (13) and lobster tails (14) . Hence.
combined with the degree of handling of these types of product.CRUSTACEAN SHELLFISH old. 19). Researchers have demonstrated that inoculated shrimp supported toxin production by C. therefore. 4. the pathogen can grow at temperatures as low as 3 °C. aureus is not a member of the intrinsic flora associated with crustacean shellfish. which was eventually attributed to a ship discharging its bilge into the coastal waters of Peru. V. Crustaceans have been linked with food intoxications attributed to Staph. is widely distributed in aquatic environments.5.g. cholerae O1 and V. aureus because of the ubiquity of this organism in the processing environment. pasteurisation and cooking. 24. GMP should be followed in the processing plant to ensure that the final product is held at 3 °C. Crabs are covered in a chitin exoskeleton. botulinum type E at 10 °C but not at 4 °C (20). cholerae non-O1. no incidence of foodborne illness caused by L.3 Staphylococcus aureus Staph. cholerae.5 Vibrio spp. 85 . whilst boils and cuts are also common sources of the pathogen. Vibrio cholerae. An epidemic that began in South America in 1991. botulinum type E is commonly associated with fish and fishery products. 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 on their hands. e. To date. pregnant women and the immuno-compromised.70% of healthy individuals carry toxin-producing strains of Staph. However. The main source of contamination is the hands of processing personnel. the causative agent of cholera. 25). so is able to survive heat treatments that eliminate vegetative cells. monocytogenes has been attributed to crustaceans. 4. although more recently the non-O1 type has also been demonstrated to cause foodborne illness in crustaceans. 22) as well as from undercooked crustacea when harvested from contaminated waters (23.5. 4. Vibrio parahaemolyticus and Vibrio vulnificus are species that have been reported to be associated with raw crustaceans harvested from estuarine waters (21. Once the competing microflora has been removed by a heat treatment. cholerae is divided into two groups. The former. which can become colonised by V. although contamination by types A and B may occur from the environment. about 50 .4 Clostridium botulinum C. as part of the natural bacterial flora. particularly crabmeat (18. especially in products that have been vacuum packaged. The pathogen has resistant spores. Control should be achieved by promoting GHP and staff training programmes.5. was associated with seafood consumption. V.
they have not been confirmed to be causal agents of foodborne illness associated with crustaceans (28). However. Norovirus. 4. 32). vulnificus is an invasive and lethal pathogen associated with wound infections. It is more commonly implicated in countries where crustacean shellfish are eaten raw. whilst 5. or freezing at a temperature of -23 °C for 7 days (30. jejuni was isolated from 36 (15%) of 240 samples of freshly hand-picked blue crab (Callinectes sapidus) meat. the potential hazard exists as C.8 Parasites The use of animal excreta in aquaculture has also been associated with foodborne parasitic infections of crustaceans.7 Aeromonas hydrophila and Aeromonas sobria A.2 °C) so that GHP coupled with controlled low temperature storage is essential to prevent the pathogen from reaching levels that may present a hazard. Japan. Parasitic infections are controlled by either cooking infected material prior to consumption. particularly trematodes (6). a marine halophile. South America. non-B Hepatitis viruses (31).5. sobria are known to be fish pathogens and are commonly isolated from ponds used for aquaculture. been implicated in a case of foodborne infection associated with crustaceans. 31. to date. coli. hydrophila is able to grow well at refrigeration temperatures (0 .30 MPN/g) (27).g. Caliciviruses. 4. Quantitative levels were below limits of detection in all cases (<0. Of these. occurs in crustaceans from warm environments. in a survey carried out in the United States between 1977 and 86 .8% of samples contained C. 4. hydrophila and A. however. The organism is widespread in estuarine waters and is a common isolate from harbour water in warm climates. e. Astroviruses and non-A. V. hydrophila has been isolated from prawns (29).9 Viruses Crustaceans grown in waters contaminated with sewage may become carriers of viruses such as Hepatitis A. jejuni has not. However.5. 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. A. Hepatitis A virus presents the most serious hazard. although A. but has the potential to cause fatal foodborne illness (26). 4. V.6 Campylobacter jejuni and Campylobacter coli C.FISH AND SEAFOOD and caused major economic losses to South American countries that export shrimp to the United States.5.5. parahaemolyticus.
A summary of microbiological criteria for crustacean shellfish is listed below. produced or imported into the European Union (EU). or chlorine concentrations similar to those found in drinking water. The risk of such hazards is far higher with fin fish and molluscan shellfish. Therefore any microbiological criteria need to be part of this HACCP-based system to verify that the system is under control. 87 . Provisions laid down concerning the use of additives would also apply to these types of products. 4. 39). although it is one of the European Union (EU) legislated allergens. which results in a persistent and apparently permanent loss of short-term memory (36). 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.11 Chemical pollutants and biotoxins Researchers have demonstrated that crabs feeding in waters contaminated with algae do have potentially high levels of algal toxin domoic acid (DA) in their viscera. are included in the Commission Regulation (EC) No.1 EU Legislation Microbiological standards for cooked crustaceans and shellfish.6 Published Microbiological Criteria The global legislative requirement for controlling the safety (and quality) of crustacean shellfish is to apply a preventive. 2073/2005. only 0. 4. 38. from farm to fork (37. which could be transferred to the meat during unskilled removal from the shell (35).47% of outbreaks attributed to Hepatitis A virus were attributed to a foodborne origin (33). 4. Norwalk viruses can also withstand heating to 60 °C.CRUSTACEAN SHELLFISH 1981.5.6. consistent with that in the stomach. 4.5. They can survive low pH levels. No foodborne incidents have been attributed to this source to date. 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. Selected parts from this regulation is presented below.
FISH AND SEAFOOD
184.108.40.206 Microbiological criteria applicable to the production of cooked crustaceans (Commission Regulation (EC) No 2073/2005 on microbiological criteria for foodstuffs) 220.127.116.11.1 Chapter 1 - Food safety criteria
L. monocytogenes Prepared crustacean products fall into the category described in the regulation as “Ready-to-eat foods available to support the growth of L. monocytogenes other than those intended for infants and for special medical purposes”. During capture and preparation there is a zero tolerance for the presence of L. monocytogenes on the product. However, to take into account the issues of natural habitats being contaminated with low levels of this pathogen there is a slight loosening of the regulation at the point of sale and for the duration of the products shelf life (refer to Table 9.I, Food Category 1.2). If the level of L. monocytogenes should be above this level then the consignment is considered to be unsatisfactory. Salmonella All cooked crustaceans and molluscan shellfish should be free from Salmonella at the point of sale and for the duration of their shelf life (refer to Table 9.I, Food Category 1.16). Should any sample test positive for Salmonella then the product is declared to be unsatisfactory. 18.104.22.168.2 Chapter 2 - Process hygiene criteria
Process hygiene criteria for E. coli and coagulase-positive staphylococci in shellfish and shucked products of cooked crustaceans and molluscan shellfish are laid down (refer to Table 9.II, Food Category 2.4.1). 4.6.2 International requirements
International requirements are based upon having a risk-based, preventive management system in place at all stages of the supply chain. The requirements for this preventive approach is described as follows (37): • Fish and fishery products should be prepared in plants certified by the local competent authority. All certified plants should comply with the GHP (39); • The fisheries industry should take responsibility, implement and maintain safety management systems based upon HACCP; • The national competent authority is responsible for the certification of fish processing and manufacturing plants, verification of effective systems, and
issuing of certificates of compliance for export products. This includes auditing and inspection programmes; • National surveillance and monitoring programmes should be in place to demonstrate that all identified hazards are under control, e.g. biotoxins, and to identify potentially emerging hazards; Within this requirement is the flexibility for countries and trading blocks to set specific standards required to deliver an equivalent requirement with those at national level. Such standards need to be science-based, especially if they are more stringent that those recommended by the Codex Alimentarius Commission. Verification that GHP have been followed during the handling and preparation of crustacean shellfish for consumption is usually done by assessing the level of E. coli and/or coagulase-positive staphylococci in the products. Legal requirements generally require that these groups of indicator organisms are present in levels <100 cells/gram of product. Pathogenic bacteria such as Salmonella spp. should be absence is 25 grams multiplied by the number of subsamples tested. 22.214.171.124 United States The FDA brought into force a Federal Mandate Seafood Rule in 1995 which forms the foundation for good practices required for the handling and processing of imported fish and fishery products into the U.S. The best practices include GHP and HACCP (37). All fish and fishery products produced in and imported into the U.S. after 17 December 1997 are required to demonstrate that HACCP principles were applied to their handling and processing. The USFDA has a zero tolerance for Salmonella in fish and fishery products; this includes farmed shrimp. The US FDA considers that the presence of pathogenic microorganisms in seafood is an indicator of adulteration (32). 4.7
Encarta 1997. Encarta 97 Encyclopedia World English Edition. Microsoft. The International Commission for Microbiological Specifications for Food. Fish and Fish Products, in Microorganisms in Foods 6. Microbial Ecology of Food Commodities. Ed. The International Commission for Microbiological Specifications for Food. New York, Kluwer Academic/Plenum Publishers. 2005, 174-249. Cann D.C. Bacteriology of shellfish with reference to international trade, in Handling, Processing and Marketing of Tropical Fish. Ed. Tropical Products Institute. London, Ministry of Overseas Development. 1976, 377-94. D’Aoust J.Y., Gelinas R., Maishment C. Presence of indicator organisms and the recovery of Salmonella in fish and shellfish. Journal of Food Protection, 1980, 43, 679-82.
FISH AND SEAFOOD 5. 6. 7. 8. 9. 10. 11. Fraiser M.B., Koburger J.A. Incidence of salmonellae in clams, oysters, crabs and mullet. Journal of Food Protection, 47, 343-5. Reilly P.J.A., Twiddy D.R., Fuchs R.S. Review of the occurrence of Salmonella in tropical shrimp. FAO Fisheries Circular No. 851, 1992, (FIIU/C851). Reilly P.J.A. Salmonella and Vibrio cholerae in brackishwater cultured tropical prawns. International Journal of Food Microbiology, 1992, 16, 293-301. Twiddy D.R. Antibiotic-resistant human pathogens in integrated fish farms. ASEAN Food Journal, 1995, 10, 22-9. Olsufjew N.G., Petrow V.G. Detection of Erysipelothrix and Listeria in stream water. Zhurnal Mikrobiologii Epidemiologii i Immunobiologii, 1959, 30, 89-94. Anon. First Listeria finding in crabmeat confirmed by FDA. Food Chemistry News, 1987, 29 (14), 38. Ryser E.T., Marth E.H. Incidence and behaviour of Listeria monocytogenes in fish and seafood, in Listeria, Listeriosis and Food Safety. Eds. Ryser E.T., Marth E.H. New York, CRC Press. 2007, 617-54. Anon. Mexican crabmeat, Greek pasta automatically detained. Food Chemistry News, 29 (19), 43. Anon. FDA checking imported domestic shrimp, crabmeat for Listeria. Food Chemistry News, 1987, 29 (24) 15-7. Archer D.L. Review of the latest FDA information on the presence of Listeria in foods. WHO Working Group on Foodborne Listeriosis, Geneva, Switzerland. 1988, 15-9. Anon. FDA to sample shrimp for Salmonella, Listeria. Food Chemistry News, 1988, 30 (36), 9-10. Anon. Monitoring of changes to listeriosis problem urged. Food Chemistry News, 1989, 31 (20), 13-7. Richmond M. Report of the Committee on the Microbiological Safety of Food. London, HMSO. 1990, 133-7. Swartzentruber A., Schwab A.H., Duran A.P., Wentz B.A., Read R.B. Jr. Microbiological quality of frozen shrimp and lobster tail in the retail market. Journal of Food Protection, 1980, 40, 765-9. Wentz B.A., Duran A.P., Swaartzentruber A., Schwab A.H., McClure F.D., Archer D., Read R.B. Jr. Microbiological quality of crabmeat during processing. Journal of Food Protection, 1985, 48, 44-9. Garren D.M., Harrison M.A., Huang Y-W. Clostridium botulinum type E outgrowth and toxin production in vacuum-skin packaged shrimp. Food Microbiology, 1994, 11, 467-72. Molitoris E., Joseph S.E., Krichevsky M.I., Sindhuhardja W., Colwell R.R. Characterisation and distribution of Vibrio alginolyticus and Vibrio parahaemolyticus isolated from Indonesia. Applied and Environmental Microbiology, 1985, 50, 1388-94.
12. 13. 14.
15. 16. 17. 18.
CRUSTACEAN SHELLFISH 22. Varma P.R.G., Iver T.S.G., Joseph M.A., Zacharia S. Studies on the incidence of Vibrio cholera in fishery products. Journal of Food Science and Technology, 1989, 26, 341-2. Madden J.M., McCardell B.A., Morris J.G. Jr. Vibrio cholerae, in Foodborne Bacterial Pathogens. Ed. Doyle M.P. New York, Marcel Dekker. 1989, 525-42. Twedt R.M. Vibrio parahaemolyticus, in Foodborne Bacterial Pathogens. Ed. Doyle M.P. New York, Marcel Dekker. 1989, 543-68. Oliver J.D. Vibrio vulnificus, in Foodborne Bacterial Pathogens. Ed. Doyle M.P. New York, Marcel Dekker. 1989, 569-600. Pearson A.M., Dutson T.R. Quality Attributes and Their Measurement in Meat, Poultry and Fish Products. Advances in Meat Research Series. Volume 9. London, Chapman & Hall. 1994. Reinhard R.G., McAdam T.J., Flick G.J., Croonenberghs R.E., Wittman R.F., Diallo A.A., Fernandes C. Analysis of Campylobacter jejuni, Campylobacter coli, Salmonella, Klebsiella pneumoniae and Escherichia coli 0157:H7 in fresh handpicked blue crab (Callinectes sapidus) meat. Journal of Food Protection, 1996, 59 (8), 803-7. Buchanan R.L., Palumbo S.A. Aeromonas hydrophila and Aeromonas sobria as potential food poisoning species: a review. Journal of Food Safety, 1985, 7, 15-29. Palumbo S.A., Maxino F. Williams A.C., Buchanan R.L., Thayer D.W. Starch ampicillin agar for the quantitive detection of Aeromonas hydrophila in foods. Applied and Environmental Microbiology, 1985, 50, 1027-30. Wekell M.M., Manger R., Colburn K., Adams A., Hill W. Microbiological quality of seafoods: viruses, bacteria and parasites, in Seafoods: Chemistry, Processing Technology and Quality. Eds. Shahidi F., Botta J.R. Glasgow, Blackie Academic and Professional. 1994, 196-219. Schantz J., McKerrow J.H. The dangers of eating raw fish. New England Journal of Medicine, 1989, 320, 1143-5. Ahmed F.E. Seafood Safety. Seafood Safety. Committee on Evaluation of the Safety of Fisheries Products, Food and Nutrition Board, Institute of Medicine. Washington D.C., National Academy Press. 1991. Blackwell J.H., Cliver D.O., Callis J.J., Heidelbaugh N.D., Larkin E.P., McKercher P.D., Tayer D.W. Foodborne viruses: their importance and need for research. Journal of Food Protection, 1985, 48, 717-23. Kapikian A.Z., Chanock R.M. Norwalk group of viruses, in Virology. Eds. Fields D.M., Knipe D.M. New York, Raven Press. 1990, 359-77. Horner R.A., Postel J.R. Toxic diatoms in western Eashington waters (US west coast). Hydrobiologia, 1993, 269/270, 197-205. Perl T.M., Bedard M.S.M., Kosatsky T., Hockin J.C., Todd E.C.D., Remis R.S. An outbreak of toxic encephalopathy caused by eating mussels contaminated with domoic acid. The New England Medical Journal, 1990, 322, 1775-80.
23. 24. 25. 26.
34. 35. 36.
FISH AND SEAFOOD 37. Ababouch L., Gandini G., Ryder J. Causes in Detentions and Rejections in International Fish Trade. FAO Fisheries Technical Paper 473. Rome, Food and Agriculture Organisation of the United Nations. 2005. Australian Government. Australian Quarantine and Inspection Service Hazard Analysis and Critical Control Point (HACCP). A Guide to Compliance with Export Control (Fish and Fish Products) Orders 2005. FAO/WHO. General Hygiene Requirements. Rome, Food and Agriculture Organisation of the United Nations. 2003.
Further Reading Spoilage and pathogens
Gram L., Huss H.H. Fresh and Processed Fish and Shellfish, in Microbiological Safety and Quality of Food Volume 1. Eds. Lund B.M., Baird-Parker A.C., Gould G.W. Maryland, Aspen Publishers Inc. 2000, 472-506. Ababouch L., Gandini G., Ryder J. Causes of Detentions and Rejections in International Fish Trade. FAO Fisheries Technical Paper 473. Rome, Food and Agriculture Organisation of the United Nations. 2005. International Commission on Microbiological Specifications for Foods. Fish and fish products, in Microorganisms in Foods 6. Microbial Ecology of Food Commodities. Ed. International Commission on Microbiological Specifications for Foods. London, Blackie Academic and Professional Press. 1998, 130-89. Sutcliffe P., Disney J. Handling, Processing and Marketing of Tropical Fish. London, Tropical Products Institute. 1977. Connell J.J. Control of Fish Quality. Oxford, Fishing News Books. 1995 Ward D.R., Hackney C. Microbiology of Marine Food Products. New York, Van Nostrand Reinhold AVI. 1991 Pearson A.M., Dutson T.R. Quality Attributes and their Measurement in Meat, Poultry and Fish products. Advances in Meat Research Series. Volume 9. London, Chapman & Hall. 1994.
4.8.2 Pathogens - general
International Commission on Microbiological Specifications for Foods. Microorganisms in Foods 5. Characteristics of Microbial Pathogens. Eds. International Commission on Microbiological Specifications for Foods. London, Blackie Academic and Professional Press. 1996. Varnam A.H, Evans M.G. Foodborne Pathogens - An Illustrated Text. London, Wolfe Publishing Ltd. 1991. Doyle M.P. Foodborne Bacterial Pathogens. New York, Marcel Dekker. 1989.
This can be called ‘kench’ curing and is mainly used for non-fatty white fish. dependent on the country or the type of product. where the salt replaces moisture in the fish flesh. and the extracted moisture is allowed to drain away. drying and smoking processes. split and opened out flat. SMOKED AND DRIED FISH Dr. It includes the salting.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. rather than for preservation itself. Simon Derrick The Grimsby Institute Humber Seafood Institute Europarc Grimsby DN37 9TZ United Kingdom 5. The fish is immersed in a brine solution in which different salt concentrations can be used according to the process. alternating fish layers with a layer of salt. their combinations and/or variations. and a preliminary operation to some smoking. CURED. 93 . and before significant spoilage takes place. Salting is both a method of preserving fish. The processes take anywhere from a few hours to a few days or even months. and salting is done by stacking the fish in layers. Fish curing is practised worldwide. and marinating processes. The fish is headed. to assure a salt concentration in the fish flesh that is appropriate for flavouring purposes prior to the use of other preservation techniques. Fish can be salted in three different ways or their combination: Dry salting.5. Wet salting or brining. drying. and is sometimes known as ‘pickling’. fish is immersed for a few hours in a relatively low concentration brine. The immersion in concentrated brines for long periods is usually used for fatty species and for longer-term preservation. both to preserve and to create a variety of new products. Usually.
depending on the type of product. are added to the marinade. under normal pressure. Drying is the preservation of fish by removing the moisture content of the fish flesh by evaporation. Fish in contact with refrigerated plates freeze. Heat is transferred to fish from heated air or a heated surface. thus delaying spoilage for a few days. This is a mixture of a dry and a wet salting process. Vacuum drying. this was done naturally by exposure of the fish to the air. this ‘brine’ can be collected and placed again over the fish several times. This is the common method of drying fish. thus reducing the water activity of the product. Smoking is an ancient process for preserving and flavouring food. Freeze drying. There are some marinating processes where salt is used for the first cure and a firmer product is obtained. The product is dried by contact with heated surfaces or by radiation. Salt-boiling. spices). It is not commonly used for fish products. or boiling it with dry salt and water. Longer shelf life is achieved in the products when salt is added. as concentrated brines are formed. as well as other flavourings (sugar. the product rehydrates quickly but does not retain reabsorbed moisture on cooking. such as ‘gravad’ fish. it can also be named ‘pickling’ or ‘blood pickling’. Food products can be dried in three different ways: Air or contact drying. the ice sublimes and the vapour is removed from the fish by vacuum pumps. It has found limited application with fish. Usually. Marinades are acidified brines. Thus. After the freezedrying process. As the water is drawn out of the flesh by the salt a brine is formed. evaporated water is removed by vacuum pump. without other preservation methods applied after boiling. 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 . Salted-boiled fish are the products obtained on either boiling fish in a brine. When the process is running in a closed system. The fish is mixed with dry salt but in a watertight container. The principle of preservation of the first method is the inactivation of enzymes and killing the non-spore-forming organisms. thus making the product unsuitable for microbial growth. the preservative effect is due to the low pH used in the process. The products are usually consumed raw.FISH AND SEAFOOD Dry / wet salting. is used as a preservative instead of salt. Marinating or pickling. where an organic acid. and the air movement removes the vapour. Traditionally. Included in this group are some lightly preserved products where no acid is added to the marinade. Salt. and the fish can be immersed in such brines for long periods. prior to or during the boiling process. usually vinegar.
the fish is subjected to heat for a period that does not coagulate protein. leaving only elevated levels of carbon monoxide. Also. temperatures below 30 °C are used. by exposure to the wood or sawdust smoke. the fish is subjected to heat for a sufficient period of time to coagulate protein throughout the fish. The different spices used in the immersion brines are also a potential source of microorganisms. SMOKED AND DRIED FISH of smoke. the salt used in the processes may contain halophilic bacteria. process temperatures are over 60 °C. In this process. The secondary objective is to impart sensory characteristics such as smoke colour and smoke flavour to the product. such as injection or dipping in a solution of liquid smoke. A dry or wet salting step is always used in the process before exposure to the wood or sawdust smoke. including pathogens. The process was designed so that the smoke is filtered to remove all smoke particles. Usually. 95 .CURED. or by means of smoke flavouring processes. Clear smoke. Usually. fish can be smoked by means other than the direct action of smoke. sporeforming bacteria or osmophilic moulds. or added to the brine with other flavourings. It is commonly used in place of traditional smoke. With the exception of some Asian countries. the sugar used in some products may contain yeasts and moulds as well as spore-forming bacteria. Smoking can be either traditional. and filtered to remove tars and particulate matter. or by means of smoke-flavouring processes with liquid smoke. Smoked fish is prepared by subjecting the fish to the direct action of smoke from wood or wood sawdust burning. 5. Liquid smoke is a natural aqueous condensate of wood smoke that has been aged. by exposure to the wood or sawdust smoke. or liquid smoke. which impart the flavour of smoke to fish. Hot-smoking. which irreversibly combine with the haemoglobin in the tissue to maintain the red colour. In this process. It can be injected into the product. After the salting step. However. Is an illegal method (in the European Union (EU)) employed to treat tuna to ensure a bright red colour is maintained during storage and transport. The smoke itself may contain spores of moulds.2 Initial Microflora The initial microflora are those of the raw fish from which the products are made. and the preservative effect is only slight. Smoked-flavoured fish. smoking is now used mainly to give the fish a characteristic flavour. It can be smoked either traditionally. Cold-smoking.
80º brine (184.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) 26.6 g of salt or more per litre of brine) may cause salt burn in this way. which is the percentage of salt in the whole weight of fish that is water plus protein plus fat. Thus. or salt in the water phase. The dissolution of salt in water is dependent on the temperature.2 105. Therefore. the fish is usually salted with a 70 . smoking or marinating processes.3 Processing and its Effects on the Microflora Owing to the wide variety of products. all using salting steps. sometimes the salt crystallises onto the surface of the fish. When a 100º brine is used.4 52.0 . The strength of the solution is described as degrees of saturation (brineometer degrees). and salt content. Salt in the water phase links the two conditions of moisture and saltiness. Different brine strengths are required according to the curing processes and/or the type of product.2 237. Salt concentration means the percentage of salt in the water contained in the fish. a 90º or above brine (237. When a milder brine.4 184. For smoking salmon. a 0º brine saturation means pure water.6 132. TABLE 5. only the most common processes can be described here.8 211. it is impossible to establish standardised processing stages in the curing of fish. it causes swelling because the fish absorbs water from the salt solution (1).8 79.FISH AND SEAFOOD 5.2 g of salt per litre of brine). etc. it is important to understand the concept of salt concentration. is employed.I). When working with salting.0 g of salt per litre of brine). This should be distinguished from the salt content.0 158.4% salt (Table 5. % Salt in the water phase = % salt content % salt content + % moisture x 100 Another important point to remember when wet salting is used is the concentration or strength of a brine. Salt in the water phase is the amount of salt compared with the total amount of water and salt in the fish. about 50º (132. it is important to state the temperature. processes and producers’ private recipes.8 211. A 100º brine solution means that the solution is saturated and no more salt can be dissolved. At 18 °C a 100º brine is produced when the solution contains 26.6 264.
15 .1 ppm and 10 ppm. Salting can be done in a number of different ways.35% and 0. See Figure 5. however. when the air humidity is high. and the salt has penetrated throughout the fish (3). When pure salt is used.1 Salting Salting of fish for preservation purposes has been used since pre-history. It is common for the dried salted cod in the Mediterranean countries to be made using solar salts. the fish is restacked with fresh salt after about a week. The fish is then stacked in piles of about 2 metres and the water is allowed to drain away. it tends to lead to a slight yellowing and softening of the product. producing characteristic pink patches.3. can be dried further to produce ‘klipfish’ or ‘bacalão’. if very pure sodium chloride is used for curing purposes. are recommended to help make the product whiter. but the concentration of copper and iron must be kept under 0. For a light salting. having a short section at the end of the tail. ‘Green cured’ split cod with about 60% of the backbone removed. can lead to the decomposition of the centre of thick muscles before it reaches the level of salt concentration sufficient to prevent spoilage. ‘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. This condition is known as ‘putty fish’ (4).3. when rock salt is used the ‘dun’ patches caused by moulds are prevalent (1). However. are likely to cause hydroscopicity of the product and.CURED. It takes about 3 weeks until proper salt penetration and moisture loss are achieved. 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. A ‘green cured’ product is obtained when the water content has been reduced to about two-thirds of the initial amount. for which there is a high demand in Mediterranean countries. and salt can speed this oxidation process. mainly in tropical conditions. Usually it is followed by other preservation techniques such as drying and/or smoking. and for a heavy salting a ratio of 1:3 salt:fish is used (1). addition of levels of 0. 97 . and such delays.0.1 Dry salting / Kench curing Usually. 5. and a ‘putty’ fish product is also obtained (1).1. respectively (1). respectively. On the other hand.1. which consequently inhibits the diffusion of the water from the bulk of the product to the surface. The most common fish used in the northern hemisphere is cod (Gadus morhua). Also. The calcium and magnesium impurities. This is because fatty fish turn rancid quickly. The salt used should be of food-grade quality.15% calcium and magnesium. and essentially free from iron and copper (3). lean white fish is used for this purpose. caught in North Atlantic waters. the fish is mixed with one part of salt per eight parts fish. SMOKED AND DRIED FISH 5. low in calcium and magnesium. outgrowth of halophilic bacteria may occur. which include calcium and magnesium.
90 are rarely achieved. Food-poisoning organisms.1. the process is mainly for flavouring purposes prior to the use of other preservation processes. such as 98 . eviscerated and/or split). when a light brine is used. Fig. 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.5. it appears that water activities much less than 0. Where fish are salted in their whole form. Processing of dried salted cod 5.FISH AND SEAFOOD Dry salting can also be used for small species without the need for opening the fish flat. such as Staphylococcus aureus could survive and grow at such water activities in the product. For large fish it is advisable to split it before salting. The greatest barriers to salt penetration are the skin and scales. although toxin production would be unlikely (4).2 Wet salting or brining As referred to above.1.3. Only small fish can be salted whole.
Fresh sardines.85 3.26 1. Histamine formation is of importance when using scombroid and some clupeidean species.51 5.76 4. TABLE 5.60 2. The main objective of the brining method of salting fish is to limit air penetration during processing. the fish enzymes play an important role by maturing or ripening the product.76 4.66 5.70 3.45 3.44 2.05 7. after being drained and pressed at 465 kg/m2 for 15 hours. can be cured in a saturated brine solution for 6 days and.11 4. thus. the liquid that comes out of the fish forms a brine.80 5.25 5.09 8.74 9. if split fish is dry-salted. There are several variations of this process.41 3. herrings and other fatty fish can be used. a saturated brine is used. Histamineforming bacteria do not generally grow at water activities below 0. limiting the amount of rancidity that can develop.25 5.26 2. sardines.76 4.66 5. but mackerel.98 6.41 4.5 10.88 5.76 4.90 (1).49 4. Anchovies are the main fish used for these purposes.54 5.23 8. It is then taken out and dried in the sun or in kilns until about 35% moisture in the product is obtained.23 3. See Table 5.3 days.35 4.II.5 4.78 6.98 5. with a shelf life of about 4 weeks (water activity (aw) about 0. the product obtained is called gaspé-cured (5).CURED.78) (1).17 5.00 3. Here. the fish is dry-salted. The fish are kept immersed in their own brine near saturation point for several days.16 7.76 99 .86 2.26 4. 5.96 1. and a red burgundy colour will develop in the product.11 3.23 2. but as it is placed in containers.65 2.85 3.0 2.70 6.91 2.10 1.22 6. SMOKED AND DRIED FISH smoking.3 Dry / wet salting In this process.95 3.76 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) 2.51 3.03 2.0 3.26 7. headed and gutted. For example.41 6.04 5.41 5. the water that comes out of the fish forms a brine in which the fish stays immersed for 2 . For preservation purposes.5 3.30 2.5 3.46 4.99 2.61 3.64 4. The brining step for smoking purposes also helps the product to develop a desirable glossy appearance.3. the brine is then replaced over the fish.78 2.73 1.09 8.61 3.19 3.56 6. and can be collected and replaced again over the fish.84 1.26 3.03 4. they can be stored at 28 °C.44 2.47 6.1.33 5.0 4.17 4.11 9. This is why it is the preferred method for fatty fish.98 6. with pressure being applied to the top of the container to help the brine to drain away.06 4.23 2.
As referred to above. All these products are ‘semi-preserves’. Other preservation techniques can be used for extending the shelf life of these products. and packed (3). they can be cut and cured for 3 days in a solution with 3% white distilled vinegar.7%) for as long as 3 weeks. Usually.3. containing 1 kg of salt for every 25 kg of fish. has a shelf life of several years. underlines the need for careful control (4). flavouring (sugar. placed in a marinade of lime juice.2 Salt-boiling Fish can be boiled after being dry-salted in a watertight container and heated for at least 1 hour.FISH AND SEAFOOD 5. giving a cooked appearance to the product. a Peruvian product. salt. The product is usually consumed raw with the high strength acid solution softening the bones enough for them to be eaten. Ceviche. and spices) is added to the brine. Another salt-boiled Indonesian product. However.5% 120grain distilled vinegar.30 minutes. The Indonesian ‘pindang’ is made by boiling the fish in a saturated salt solution and it is sold as is. The fish can be soaked in a high concentration of vinegar (about 4 . and lasts for 3 . and 6% salt. only the nonspore-forming organisms are eliminated in the process and. There is epidemiological evidence that these products have been the cause of some 100 . or can be boiled by being placed in a basket. Herring can also be cured for several days in an 80 to 90º brine with 2. the occasional implication of pindang in cases of foodborne illness. A much firmer product is obtained when the fish is salted prior to acidification. Afterwards. Another variation uses sour cream instead of the second brine step. Traditional jellied eels are a product where pieces of fish are cooked in boiling water.3. whereas the latter method preserves the fish for only a few days. ‘ikan kaju’. even if the product is hermetically sealed. up to 1 . it can be repacked in milder brine (1 . The lime juice plays the role of the vinegar. drying and salt-boiling steps until a hard translucent block is obtained (4). The first process can give a longer shelf life. 5. After this. In a second step. because of the salt content of the product. but it has further smoking. usually flavoured with onions and hot chillies. They will have a salt content >6% (w/w) in the water phase.4% of salt) for distribution and sale (1).2 months. or a pH <5 and they are stabilised for a limited period. herring or other fatty species are used. outgrowth of bacterial spores after processing may occur (4). then packed in barrels with a 70º brine.6 days without refrigeration (1).2% acetic acid and 2 . and even death.3 Marinating / pickling Fish can be brined in solutions based on organic acid instead of salt. Once cooled. until the flesh is soft. The cooked portions are poured into bowls containing gelatin dissolved in water (10% solution) and sometimes a small amount of vinegar. usually requiring refrigeration below 10 °C for longer-term storage. which is dipped into a boiling brine for 10 . is made of different varieties of fish. they are packed for consumption within 2 weeks at chill temperature (6).
are commonly dried in Asian countries. 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. Provided the product is kept in dry and fairly cool conditions. Stockfish products have a golden brown colour and are very hard. South America and the Caribbean. Some bacteria and enzymes are still active in the presence of acid and salt. Other products. it being preferable to use frozen salmon or to freeze the product at the end of the process to avoid parasite survival. which are placed in a mixture of salt. etc. Usually. are sundried and. 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). sea cucumbers and seaweeds. 5. the fish is dried naturally in a relatively cold environment. such as Listeria monocytogenes and Clostridium botulinum. are generally air-dried at sea. under a light pressure. a very fine delicacy for Oriental food lovers. At pH levels below 4. Fresh dill is placed between the layers of fish. by using either solar heaters or mechanical dryers. are likely to survive in these mild-processed products. while in the latter case there is a provision for fans or other means of mechanically moving the air (1).There are several products named marinades (matjes. still made without prior salting on a large scale. Dried shrimps are widely used in Asia. jellyfish. mixed with salt. In the first case. and the stacks are placed in the refrigerator. Shark pectoral fins. gravad fish. after frying. split and dried gadoid species) are traditionally made in cold climates.) where no acid is added to the process. some fish species such as moray eel and varieties of dog fish. It takes about 2 days to marinate completely. Gravad fish is a Scandinavian product made of salmon fillets.0) (7). gutted. it will last for many years (5). used as an appetiser.5. marinated products will keep for several months at a temperature of 4 °C. and a fine product is obtained. Matjes are made of fat young herring. such as squid. then mild-cured with special seasonings and packed in barrels with 80 º brine (5). They are an important dried product.4 Air drying This is probably the oldest method of fish preservation. bacterial toxins and parasites (7). They are not cooked prior to consumption and they have a limited shelf life even at refrigeration temperatures. Dried octopus is also a very common dish in the interior of the country. depending on the thickness. air flows naturally. Stockfish is extensively used in Portugal. 101 . Italy and Spain (5). It is important to assure the quality of the raw material. The most common method used for fish products is the air-drying process. the temperature and the amount of pressure applied (5). SMOKED AND DRIED FISH foodborne diseases related to the presence of biotoxins. Stockfish (dressed. and the flesh will break down. Foodborne pathogens. the fish is dried after salting or brining. sugar and spices. but sometimes it is dried without prior salting.CURED. In some coastal areas of Portugal. In Scandinavian countries.3.
and has the primary objective of creating a product with specific taste and textural characteristics. botulinum). The fish must be stored in reasonably dry conditions to avoid moisture absorption. some of which are listed in Table 5.III. They must be soaked in water. aureus) or spores (e. To prevent this. neither it. nor subsequent cooking operations will affect the presence of toxins (e. and increasing amounts of such products are making their way into the market place. more commonly nowadays. tarpaulin.g. the ‘green cured’ cod is rinsed or washed with water and then hung to dry either by the sun and breeze or. there is an increasing demand from people that have emigrated to industrialised nations for traditional products. 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.83 (9)). Italy and South America. and it is exported to Portugal.FISH AND SEAFOOD When the fish is sun-dried in tropical conditions. the product can be held for months at room temperature. by warm air circulating in a drying chamber. the fish must be dried slowly enough to avoid case-hardening. The product is dried to a final water content of about 25 to 38% (3). The amount of moisture transfer from the fish to the air is dependent on the relative humidity . if the rate of water removal is too high in comparison with the diffusion rate. toxic secondary metabolites of the moulds Aspergillus flavus and Aspergillus parasiticus are not produced at aw <0.g. Owing to restriction of fishing areas for Spanish and Portuguese vessels since the 1970s. 102 . the diffusion of the bulk water to the surface of the fish is blocked. and the water must be changed several times before cooking. When no more water can be removed from the fish. Traditional processing conditions provide ample opportunity for product contamination from a wide range of hazards and sources. Dried salted products must have a water activity below 0. the protein must be set or denatured with low-temperature drying before applying higher temperatures. It is important to balance the drying rates and quality/safety issues. Iceland and Canada. Whilst the production of dried fish products is tightly controlled in industrialised countries. the better the moisture transfer. or wooden drying rack to dry in the sun.75 (1). but fast enough to avoid spoilage caused by bacterial and enzymic activities (1). Spain. A highly demanded dried fish product in Mediterranean countries is dried salted cod. making a water barrier and consequently. Thus. In conclusion. In many such artisanal fisheries the drying is simply conducted by spreading the fish on the beach. Whilst the low aw of the final product will inhibit microbial growth. dried salted cod (bacalão) is now mainly produced in Norway. which leads to mould growth (aflatoxins. Portuguese and Spanish fisheries cannot satisfy their own countries’ demand for cod. Although controls are in place to ensure that products processed in such relatively uncontrolled conditions do not enter the supply chain. skin up. However. After dry salting. that may have contaminated the product during the drying process. spoilage and possibly C. the skin will form a hard case (case-hardening). C. the quality is more difficult to control. France. botulinum growth might occur in the bulk of the product. typically in stews and soups.the lower the humidity. Staph.
aureus and Saccharomyces cerevisiae (10). Hardwoods are considered better because of the milder flavour they impart to the food. There are many fish species that are used for smoking purposes . tuna. swordfish. temperature and humidity can be automatically controlled and the process is faster because the directed air movement draws the smoke through the fish. Staph. are some of the ways of reducing the polycyclic aromatic hydrocarbon (PAH) compounds (10). It can be used for cold or hot smoking. Blends of various woods can be formulated to produce sawdust with specific flavours. Aroma and flavour are a blend of smoke components (3). is mainly a process for flavouring fish rather than a preservation technique.5 Smoking Smoking today. 5. Proper cleaning and preparation prior to smoking and washing of the fresh or frozen fish by water spray or a continuous 103 . cod.3. Preventing the occurrence of carcinogens during smoking has been an issue for the past several years. In the mechanical kiln. haddock. The quality of the smoke is related to the type of wood used. The production of smoke by wood or sawdust burning can be done in a mechanical kiln or in traditional chimney kilns. aureus (toxin) C. as referred to above. eel. and the use of smoke generated by superheated steam or liquid smoke. botulinum (spores) Pesticides / Chemicals Physical Glass / sand etc. softwood colours the product quickly. trout. SMOKED AND DRIED FISH TABLE 5. It has been reported that the smoke from mahogany. Top-quality fish must always be used. herring. However. using electrostatic filtration of the smoke. oysters. The smoking procedures vary widely. such as Escherichia coli. making full description impossible here. cod roes. mussels and many others from tropical countries.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. clams. Maintaining the temperatures of pyrolysis between 425 and 200 °C. white mangrove and abura has an inhibitory effect on some microorganisms. mackerel.salmon. sprats.CURED.
thawing must be done slowly for 12 . it will reduce the salty flavour. However. spices. filleted. Cod roes.3. however. smoked cod fillets and finnan haddock are usually cooked before consumption.e. gutted or filleted. are boiled before smoking (12). When frozen/thawed fish is used. Sugar. it will double the drying time in the kiln to get the same total weight loss as for dry-salted fillets (11). 70 – 80 º brine is commonly used. but it will increase towards the end of the process (10). skin on or off). temperatures must be kept under 30 °C throughout the process. as it retains more moisture. but a ‘safety’ (i. after being smoked need to be chilled immediately to 4 °C before being packed. Also. which is important for the appearance of these products (4). permitted colourings or smoke flavours can also be added to the product in the brining step. Thus. canned or bottled. Some herring products. but this concentration will again depend on the type of product and the length of the brining stage. or packed in retortable foil pouches.and hot-smoking is the temperature of the process. All the smoked products. whether cold. If frozen fish is used. they can be vacuum packaged. pasteurisation) temperature is not achieved in any stage of the process. covering the fish surface completely (10). it will take up salt twice as fast as fresh fish (10).50 ppm) should always be done just before process initiation (3). 5. temperatures and times used in processing coldsmoked fish are very favourable for the proliferation of food-spoilage and foodpoisoning microorganisms. Whether the product is prepared headed. Thus. giving the product the desired appearance and flavour (4). tuna and swordfish. skin off or on. such as smoked salmon-trout.FISH AND SEAFOOD water flow system with chlorinated water (25 .5. The salting period is dependent on the size of the fish and its presentation (whole. The maintenance of temperatures below 30 °C avoids protein coagulation. Depending on the type of product.10% of the weight of the fish. an attractive glossy pellicle on the fish fillet surface is achieved after draining. thus allowing the deposit of smoke on the fish surface. the following step is salting. low temperatures will slow the uptake of salt. In cold-smoking.18 hours at 4 °C. or quick-frozen and stored at -18 °C (2). are not cooked prior to consumption. Wet salting or brining is preferable to dry-salting when smoking fillets because. the excess salt is removed with water. and the cold smoke is denser. Brining at 10 15 °C is recommended (10). all cold-smoked products should be cooked before consumption. If wet salting is chosen. usually dry-salted and cold-smoked. 104 . The fish can be dry-salted using a weight of salt 5 . Drying should be minimal at the start. The length of the smoking process for such products is much greater than for the hotsmoked fish. shrink packaged. This difference is crucial for safety issues because the flesh is not cooked. owing to the elution of soluble proteins. the highly desirable cold-smoked salmon. wooden boxes or master cartons (10). as well as many other products prepared in the same way.1 Cold-smoking The major difference between cold. Sugar can be added together with the salt.or hot-smoked.
During smoking. in order to produce toxin. but they should be cooled to a temperature of 10 °C within 3 hours. Pathogenic vibrios. Smoked products can be also packed under controlled-atmosphere packaging. an initial humidity of 70% seems to be preferable because it is a good compromise between maximum smoke adsorption and minimum casehardening (1).CURED. See Figure 5. Smoked salmon fillets are usually vacuum packed whole or after slicing. Afterwards. Vacuum packing is recommended only for coldsmoked salmon. can grow with salt levels of 10% in the water phase. Vacuum packing extends the shelf life of the lightly smoked products by suppressing the growth of other bacteria and moulds. The protein must be set or denaturated with lower temperatures before the application of higher temperatures to avoid case-hardening. A typical temperature set is as follows: firstly. they are more sensitive to sodium chloride and unlikely to grow in smoked products (13). 5.2. thus allowing any psychrotrophic C. and hotsmoked salmon (kippered) (3). Although other human pathogens that can grow at refrigeration temperature. a drying period at 30 °C.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. the products are cooled. but the proteolytic types A and B. such as Yersinia enterocolitica. as. further cooled to 3 °C within 12 hours and then packed. typically mesophilic. there is no point in the process that can fully assure the absence of L. a final cooking period at 80 °C (12).3. a partial cooking period at 50 °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). if a 105 . if present. To avoid growth of non-proteolytic C. The exception is vacuum packed fish. and their sensitivity to salt (13). stored at 4 °C and packed. can be isolated from seafood. botulinum type E the product must have at least 3% salt in the water phase and be stored below 5 °C. monocytogenes. Length of the smoking process at each stage will depend on the species and the type of product. condensation will occur inside the pack. gravad salmon (also called gravad lax or gravlax). which underlines the danger of storage at abuse temperatures of >10 °C (13). SMOKED AND DRIED FISH During cold-smoking. secondly. Hot-smoked products require a longer cooling stage.5. These products should be vacuum packed at room temperature. Furthermore. Neither the smoking temperature nor the salt content is enough to kill the organism. refrigerated storage and vacuum packaging still allow its growth (12). This temperature should be maintained during storage and distribution (3). If the refrigeration chain is maintained. are of concern when smoking fish from tropical waters (13). thirdly. On the other hand. risk from the presence of Bacillus cereus and Clostridium perfringens can also be excluded as potential hazards because of the relatively high temperatures they require to grow to. if they are taken from a chill room at 3 °C and packed.
FISH AND SEAFOOD product is packed too warm without being cooled to room temperature. it will have a reduced shelf life. Fig. Process of cold-smoked sliced salmon 106 .2. 5. and moulds will grow on the product (12).
is a boiled smoked fish product that is used in Japanese cooking to impart flavour. 22. Hon kare bushi is a form of Katsuobushi from which shavings are cut to make aromatic stocks for use in Japanese dishes (24).5. Eels. 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). and substances unconnected with smoke but with a smoky flavour or smell. Mackerel. are used. hot-smoked fish can be classified as a ‘heat-treated or pasteurised’ product (7). 23). rewetted and remoulded to give the desired shape and consistency.3 Liquid smoke Smoke-flavoured fish represents a small percentage of the total smoked fish industry output. apart from the improved appearance. such as yeast derivatives. they can be stored for 6 months at -30 °C and for even longer when frozen in vacuum packs (12). to have no further advantage over other packing methods for other fish (12). including Eurotium repens and Eurotium rubrum.3. 18).6 hours and smoked for 3 . When refrigerated at 3 °C. Vacuum packaging retards the onset of rancidity when fatty fish is used. a very expensive and fashionable smoked product in the Netherlands. The classical liquid smoke is 107 . Growth of these moulds on the tuna results in the extraction of moisture. Boiled or steamed clams. Katsuobushi. resulting in a characteristic flavour (20. are usually hot-smoked after being immersed in 80º brine or dry-salted (6). similar to masmeen. then hot smoking and finally sun drying it. They will keep well frozen. The production method is similar to masmeen. cooking in brine. chilled or freeze-thawed. are usually 80º brined for 5 . Natural smoke extracts. Aspergillus oryzae. SMOKED AND DRIED FISH There are several species that are usually hot-smoked and the process can vary markedly. The product is removed from the drying cycle. but seems. white fish about 8 days. synthetic smoke flavours. (Penicillum cyclopium and Penicillium puterellii) (19). and many closely related Aspergilli (Aspergillus ochraceus. but mechanical drying is used instead of sun drying. 5. 21.CURED. A type of dried hot smoked tuna called masmeen or masmin is produced in the Lakshadweep islands of the Indian Ocean. 17. including ‘umami’ (16. Aspergillus ostianus. The composition of commercial liquid smoke can vary widely depending on the method of manufacture. moulding it into long thick sections. Aspergillus tamarii) and several Penicillium spp. They are generally not cooked prior to consumption. fatty fish will remain in good condition for 6 days. Masmeen is usually prepared from bonito tuna by eviscerating and filleting the fish. In terms of safety. which means that any bacterial contamination and growth after the heat treatment is a serious hazard that must be controlled by Good Manufacturing Practices (GMP) and factory hygiene/sanitation after the heat treatment (7). Hot-smoked products made from white fish will usually keep better than those from fatty fish.3½ hours using the typical temperature set for hot-smoking referred to above (12). break down of fats and the moderation of smoke flavour. The product is often allowed to go mouldy by incubating it in a chamber containing a natural fungal flora.
In experiments comparing traditionally smoked fish with liquid smoke. drying. The European Scientific Commitee for Food.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. applying it as a dip after brining.6 to 2. 5. Fresh fish spoilage may be measured by 108 . botulinum types A and E spores in several fish species that were stored at 25 °C for 7 and 14 days (25).3. pickling and marinating fish were originally used to lengthen the short shelf life of fresh fish. Smoke-flavoured fish processed using liquid smoke can be prepared by including liquid smoke in the brine. it has been reported that the liquid smoke-treated product has less pronounced aroma and flavour (25).03 μg/kg of benzopyrene. nutritional neutrality and antioxidant activity (27). smoking. both of which are considered potential carcinogens. There are two groups of chemicals of concern in smoke: polycyclic aromatic hydrocarbons (PAHs) and nitrosamines. When liquid smoke was used.5. spraying is preferable because it is easiest to control flavour and acceptable with this method (3). the result of this process is seen as a manipulation of the natural degradation of tuna colour that hides the true nature of the product quality and therefore has been made illegal in the EU. The synergistic effect of liquid smoke with other preservatives could reduce the amount of salt in the product when light diets are necessary whilst still meeting the necessary safety criteria (26). It has been shown that liquid smoke can reduce the formation of PAHs (3). so as to impart an unnatural red colouration to the flesh. established in its guidelines that smoke flavourings added to the food product must assure a level lower than 0. injecting it into the fish flesh.8% and still provide protection for 7 days (25). and less than 0. According to some recent research.4 Spoilage Brining.06 μg/kg of benzoanthracene in the final foodstuff (28). Liquid smoke was considered to provide advantages of safety.FISH AND SEAFOOD smoke condensate that is dissolved in water or oil. or applying it as an atomised spray within a modern automatic smoke kiln. 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. or smoke extracts in organic solvents. 5. the salt level could effectively be reduced from 4. 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. Whilst neither the CO or the carboxymyoglobin complex are harmful in themselves. consistency of flavour and colour.
g. therefore. however. Micrococcus spp. pickling and dry-salting all introduce bacteria to the fish. For some products. these halophilic organisms may grow and cause spoilage. contaminant bacteria from other materials used to produce the product. SMOKED AND DRIED FISH grading schemes or quality indices. Pickling and/or the anchovy process uses solid salt extraction of fish fluids to form a brine solution (cure). and high concentrations are usually required to inhibit their growth fully. for some. Once placed into brines. and. Brining.000 cfu/ml of brine may build up. the pickle and dry-salting preservation processes require further processing for significant shelf life increases (e. mainly comprising corynebacteria and Gram-negative bacteria from the fish and water (29). or extended refrigerated storage. a process similar in microbiological effect to that of saturated brining. the bulk of the microflora comprises Bacillus spp. the fish are cut and splayed to allow greater surface contact with the salt. but.. Upon salt packing. a product may become undesirable for consumption as a result of changes in its sensory characteristics. Often. otherwise. also. limits the types of microorganism capable of growing. The use of drying and/or salt to lower the water activity. This is not usually the case for smoked fish as the residual salt concentration is not high enough to inhibit all the non-halophiles. contact with the bacteria 109 . Even saturated brines allow some halophilic organisms to grow. Endogenous marine fish bacteria prefer to grow in the presence of salt (marine concentration approximately 2. rock and manufactured salts did not usually contain halophilic bacteria (29). these bacteria tend to continue to dominate the flora in the liquid.CURED. chemical oxidation. Within the limits of safety. the fish are first cleaned (known as ‘rousing’) by a salt rub to remove any outer slime.3%). Spoilage needs to be carefully defined for each product. which may also be produced to describe the quality attributes and their loss over time for processed fish. both of these will affect the spoilage patterns. the fish flora are transferred to the brine mix. When acid marinating or smoking. marination or drying). Once these changes are defined. Salts used for brining have been found to carry a microbial load of between 10 and 1000 organisms per gram of wet salt. solar salts investigated carried the highest load and included halophilic organisms as a result of their production method (open-air evaporation of sea water). the shelf life may be estimated and investigation made into the causes of the loss of sensory quality. In salts. or their products require refrigerated storage. and coryneform bacteria. Usually. the production process will modify the microbial flora present on the product. as the fish are added. for others. and of smoke to add chemical preservatives. The microflora attached to the fish is relatively unaffected by light brines used before smoking. the consequent reduction in the pH will help to inhibit spoilage organisms. a very dry product is required to produce 'shelf-stability'. Some brines are re-used over a period of days and a load of 10. When heavy brining or salting is important for product shelf life. Stronger and longer brining has an effect like that of pickling. spoilage is due to endogenous microorganisms and enzymes of the fish. which are found in the salt.. Most marinated and smoked fish products are first brined or salt-pickled.
as little if any growth will occur below 5 °C on salted fish (29). the best way of controlling the organisms is to refrigerate the fish. together with a range of Aspergilli. and Halobacterium spp. A. and Aspergillus penicilloides. or after drying if the fish is stored in excessive humidity. and still produce a reasonable dried product. halophilica was isolated 110 . Therefore. This condition is often associated with case-hardening. Insufficient salt penetration into the flesh leaves areas where Gram-negative rod-shaped bacteria and micrococci grow and cause a putrefactive spoilage. Bacteria from these two genera also grow sufficiently in brine and even salt to cause pink discolouration. lactic acid bacteria (LAB) and moulds. it is possible to wash off the infected parts. most notably Penicillium citrinum and Penicillium thomii. 34) have shown that P. which.. yeasts. spore-forming bacteria. protein breakdown.g.FISH AND SEAFOOD in the salt is made and the flora will develop accordingly in the blood pickle. ‘pink’ and ‘dun’. niger. The metabolism of the organisms will produce off-odours. powdery growth. The microbiology of heavily salted products is dominated by Gram-positive. spoilage of these pickled and dried fish in refrigerated storage tends to be due to chemical changes (textural changes due to proteolysis. The bacteria are osmosensitive and will lyse in water. 32. like the ‘pink’ organisms. though only the latter two species were considered likely to have grown on the fish (32). Their growth produces noticeable spoilage of the fish appearance but is also controlled by refrigeration. may also be found in the salt used in production. W. Only halophilic microbes may grow. Basipetospora halophila. although it may occur in other salted fish. or fat breakdown) caused by enzymes rather than by microbial growth. it was found in about 50% of samples examined. sebi is considered to be the main spoilage agent of dried fish produced in temperate regions (30). Sporendonema sebi)). pisce was the main fungus isolated. Aspergillus wentii. ‘Sliming’ or ‘putty’ fish is a condition that occurs in poorly or insufficiently salted cod (e. A number of spoilage organisms can grow and produce conditions such as ‘sliming’. gaspé). 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). sometimes covering the whole of the fish with a whitish. (syn Oospora halophila) and Polypaecilum pisce are two white halophilic moulds that are associated with spoilage of salted fish (31. At the pre-drying stage. yeasts and fungi. The bacteria form coloured colonies on the surface of salted fish prior to drying. (syn. Unlike ‘pink’. rubrum are also common isolates from dried fish. ‘dun’ is not characterised by the production of offensive odours. rancidity due to lipolysis. B. 33). as the water activity of saturated brine solutions is low enough to inhibit most bacteria. halotolerant or halophilic micrococci. Studies conducted on dried and salted fish from Indonesia (31. ‘Pink’ describes a condition associated with surface growth of the pigmented halophilic bacteria Halococcus spp. Several Penicillium spp. were also isolated. including A. E. repens and E. including hydrogen sulphide (26). flavus. The condition describes the growth of the halophilic moulds (Wallemia sebi. if not too contaminated. Eurotium amstelodami. Aspergillus sydowii.
It has been reported that the type of salting (dry. The salt in the water phase must be high enough to inhibit the growth of C. but often colour changes due to chemical oxidation of the smoked product will limit the shelf life. The preservative effect of the smoke will inhibit some spoilage microorganisms.5% sodium chloride in the water phase will inhibit some bacteria. flavus. most marine bacteria are moderately halophilic and are not effectively inhibited by these low salt concentrations. 33. so the bacteria spoiling these products will be psychrotrophic or psychrophilic.CURED. as already stated. has been isolated from dried fish (35). able to grow on dried salted fish. which does not create the salty conditions required for spoilage by those halophilic microorganisms. Fish processed by cold-smoking. Refrigeration is necessary. but. For more information on spoilage see Table 5.747. around the water activity of saturated sodium chloride. but this will not prevent the growth of spoilage bacteria. not the growth of spoilage organisms.IV. In Vietnam. SMOKED AND DRIED FISH infrequently at low levels.V gives details of pH ranges and aw minima for the growth of some important spoilage microorganisms and pathogens (38). Vacuum packing limits chemical oxidation. 36). Marine vibrios were the dominant flora in smoked salmon that was drysalted. Reports for vacuum packed cold-smoked salmon have identified more than one organism associated with spoilage. botulinum type E. with a lower aw limit for growth of 0. Cold-smoked fish will contain microbial flora representative of that of the raw fish. so the most probable spoilage organisms will be spore-formers or post-smoking contaminants. The brining to 3 . Aspergillus spp. it was. where a high internal temperature is reached. or any smoking process where temperatures in the interior of the fish do not reach >50 °C for a significant period of time. Smoked fish products tend to have a light brine treatment. 111 . Pseudomonadaceae. will have a different residual post-smoking flora from 'hot-smoked' fish. LAB. Aspergillus clavatus and Aspergillus niger. however. including A. Technical studies demonstrate the ability of many of the xerophiles above to grow at typical water activities seen in dried salted fish (32. whereas a mixture of Enterobacteriaceae and LAB were prevalent in brineinjected salmon (37). Table 5. Refrigeration of hot-smoked fish will greatly reduce the number of organisms able to grow and will lengthen the shelf life.. Vibrionaceae and Enterobacteriaceae may all play some part in shelf life limitation. and microbial spoilage is more likely. The heat during hot-smoking will kill vegetative microorganisms but not all spores.or wet-salting) in the cold-smoked salmon process leads to different microflora growth at the onset of spoilage.
W. Histamine producers Halophilic spoilage organisms Spore formers and PPC None Acid-tolerant spoilage organisms Xerophilic moulds Fresh fish spoilage organisms. 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. Dun. sebi and B. + to +++ strength of preservative effect 4 Post-process contaminants 112 .not used/unchanged. boiled. 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. . halophila Halophilic spoilage organisms Halophilic spoilage organisms.FISH AND SEAFOOD TABLE 5. Enterobacteria and PPC Spore formers and PPC Variable Salt marinades Salted then boiled Brined then boiled Salted then dried Canned Fish Gravad. and Halobacterium spp. Matjes v3 + ++ v - +++ ++ ++ - v +/- ++ ++ ++/++/- Pindang Bacalao + +++ Gaspé cure Anchovie ++ Anchovies ++ - - - ++ ++ Saturated brined fish Salted then boiled Salted. LAB.variable.IV Spoilage of salted.
8 1. the fermented products being the most dangerous group (7).9 0.5 0-11 3.61 Organisms Most Pseudomonadaceae C. The presence of salt in cured products has a great effect on the growth of the bacteria.97 0. A number of surveys have shown that fish from temperate waters is mainly contaminated with the psychrotrophic C. but if the fish products are stored at temperatures <10 °C. the importance of cold-chain integrity must be emphasised.5-10 4. temperature ≤5 °C and salt ≥3% in the water phase is sufficient to prevent any growth of type E. although the combination of the two factors. the non-proteolytic type E strains can grow in up to 5% sodium chloride.8 0. SMOKED AND DRIED FISH TABLE 5. pressure on the traditional methods of curing. Sodium nitrite in conjunction with sodium chloride is known for its ability to inhibit C.5.5% of salt in the water phase and great care must be taken to avoid any temperatures >10 °C (13). sodium 113 . botulinum. and as botulism is a serious hazard in foods. The concentration required to prevent growth at room temperature can vary from as low as 3. when present in the raw material. botulinum type E Staph.75 0. Examination of 165 outbreaks of botulism caused by fish products showed that smoked. It has been shown that. popular demand for healthy foods has led to a change in the tastes preferred by the public and. 3% sodium chloride in the water phase is enough to inhibit growth of type E for at least 30 days (39).5-10. and C.5 4.5 5. with this change. but the concentration of salt in smoked salmon or trout (usually varying from 1 4% salt) is not high enough to prevent growth. but the presence of the mesophilic types A and B is of concern in fish from warmer waters (13). the products were heavily salted.5-8.97 0. Since neither the smoking nor the drying parts of this curing process are particularly severe.86 0. types A and B are not affected by 3 .V pH ranges and aw minima for growth of microorganisms (38) Organisms Most Pseudomonadaceae C. under optimal conditions.5% to 5%.0-9. respectively. salted and pickled products accounted for 10. In the United States. 9 and 8 of the outbreaks. botulinum Staph. Before. botulinum type E and production of toxin. aureus Most spoilage bacteria Most spoilage moulds Halophilic bacteria Xerophilic moulds Aw 0.5-8. botulinum type E. aureus Yeasts Moulds Lactic acid bacteria pH 5. could not grow and produce toxin. However.5 5. although approximately 10% salt in the water phase is necessary to fully inhibit the mesophilic types A and B strains (14).1 Pathogens: Growth and Survival Clostridium botulinum In the last few decades. smoked and dried.CURED.
The degree of concern in coldsmoked salmon is low unless temperature abuse >10 °C occurs (13).0% salt in the water phase (30). The FDA established that hot. when the bacterial load of the product is lower. gills and guts as well as in several lightly preserved fish 114 . 5. monocytogenes is a ubiquitous organism that has been detected in fresh fish skin. but. An outbreak of Shigella in an Italian cruise ship.2 Staphylococcus aureus This is a ubiquitous organism but its main reservoir and habitat is animals and. but requires higher temperatures for toxin production (>10 °C).30% of the population positive for enterotoxin-producing strains (41). aureus growth. if it contains sodium nitrite ≥100 ppm. Thus.5. It is a halotolerant bacterium able to grow at water activities as low as 0. has been reported (42). 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. they will grow rapidly (7). aureus. after processing. Once the product is contaminated.94 (7). coli all occur on fish products as a result of contamination from animal/human reservoirs.5% salt in the water phase. 5.5. but.4 Listeria monocytogenes The psychrotrophic L. they do not grow well in the raw material. Usually.5.4. The human carrier rate can be as high as 60% of healthy individuals with an average of 25 . As with other organisms. its occurrence in a final product is due to contamination during processing by an infected asymptomatic carrier with poor personal hygiene. if storage temperatures are over 10 °C. the organism can survive most of the light curing processes. As they are poor competitors. However. it should contain not less than 3.5. growth and toxin formation may occur. if recontamination occurs and storage conditions are favourable.3 Enterobacteriaceae The pathogens Salmonella.or cold-smoked fish should have at least 3. and the minimum pH for growth is 4. In foodstuffs.86 and 10% sodium chloride. the human nose. as they require more handling. 5.0 . related to the consumption of a raw smoked fish meal. are particularly susceptible to contamination and. the risk of infection can be eliminated by proper cooking. The organism is mesophilic with a minimum growth temperature of 7 °C. they can survive lightly cured processes when the sodium chloride content is less than 5% and water activities are higher than 0. if the person handling the product is a carrier of Staph.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. Shigella and E. throat and skin. Infection by Shigella sonnei has increased in the UK. more importantly. the organism may contaminate the product. Sliced smoked fish products.
The round worms. The use of LAB in fish preservation is a topic that has been studied in projects (UP. have been shown to have an anti-listerial activity in many food products. This includes matjes herring. Vacuum packaging (VP) of sliced smoked fish creates a microaerophilic atmosphere. SMOKED AND DRIED FISH products such as cold-smoked fish. salt levels of 8 . do not survive 1 min at 60 °C. As they are heat-sensitive. favours growth of psychrotrophic LAB.92 . cold-smoked fish. the CT 3162-FAIR program. They have complicated life cycles. a round worm. and in CT95-1207FAIR). a number of fish products. 5.12%. in combination with curing agents and refrigerated storage. has also been detected in cod (7). Two outbreaks in the United States have been reported to be linked to salmon (10).514. from freshwater fish.95 (45). in the EU Fisheries R&D-FLAIR program. both being detected in cold-smoked salmon. Unlike some European countries. such as ‘lightly preserved’ fish products. which considered L. As its presence has been reported in lightly preserved fish products whose processing chain cannot fully assure the absence of the organism. the only concern to public health is when eating raw or uncooked fish products. monocytogenes in levels at or above 102 per gram (46). from seawater fish. Some LAB. when an outbreak of nine cases of listeriosis was suspected to have been caused by gravad or cold-smoked trout (47). widely used as starter cultures in food processes. monocytogenes pathogenic for only specific segments at risk.5. Thus.5 Parasites Despite the presence of parasites in fish being very common.CURED.0.the round worm Anisakis spp. Marine LAB is part of the natural flora of chill-stored vacuum packed salmon and cod. which include intermediate hosts. Of microbiological concern in the northern hemisphere are two parasites . are considered unsafe. lightly salted caviar. monocytogenes’ in 25 g of the finished product. marinades (ceviche) (43) and gravad fish (44). pH 4. and other local traditional products (7).6 and water activities below 0. 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. a short period of freezing either of the raw material or of 115 . which means that cooking a fillet 3 cm thick for 10 min at 60 °C will kill any worms present (48). Until 1997. and the tape worm Diphyllobothrium spp. the USA established ‘zero tolerance’ for the organism.2. As freezing to -20 °C and maintaining this temperature for at least 24 h will kill all the nematodes.8 . there were no reported outbreaks related to cured fish. researchers are looking for other ways of preventing the pathogen’s occurrence or growth. most of them are of little concern to public health. Pseudoterranova dicipiens. which means that the process must conform to a level of ‘no detectable L. Its limiting conditions for growth are 0 . which. gravad fish.45 °C. or nematodes.9.
If the Bacillus counts exceed 104/g. 5. 48h ± 2h) Indicator organisms‡ Enterobacteriaceae E. salami.<100 20 . soft cheese. These foods fall into category 5.VI Guidelines for the microbiological quality of ready-to-eat smoked fish. cholerae Vibrio parahaemolyticus L. perfringens B.<104 100 . monocytogenes Staph.FISH AND SEAFOOD the final product should therefore be included in the processing as a means of controlling parasites (7). aureus C. E. texture.<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.<104 20 . coli (total) Listeria spp. † Guidelines for aerobic colony counts may not apply to certain fermented foods. cereus and Bacillus subtilis# <20 <20** <20 <10 20 . coli O157 & other VTEC Vibrio.<103 N/A 100 . (not L.<100 200 . and the levels or absence of indicator organisms or pathogens. not detected in 25 g Campylobacter spp.<104 104 .<104 present in 25 g ≥103 >100 ≥104 ≥104 <103 103 . the organism should be identified # N/A denotes not applicable 116 .<100 20 . ‡ On occasions some strains may be pathogenic.6 Published Microbiological Criteria TABLE 5. taramasalata and cooked shellfish sampled at the point of sale (adapted from (11)) Criterion Satisfactory Borderline limit of acceptability (cfu/g unless stated) Unsatisfactory Unacceptable/ potential hazard* (cfu/g unless stated) (cfu/g unless stated) Aerobic plate count† (30 °C.<100 10 . smell. for example. monocytogenes) (cfu/g unless stated) <106 106 .<107 ≥107 N/A <100 <20 100 .<100 >104 >100 N/A N/A <20 20 . Acceptability is based on appearance.100 >100 N/A Pathogens Salmonella spp. and unpasteurised yoghurt.
Torry Advisory Note No.H. Moody M. 82 (revised). Wareing P. 1997. New York. 1982. Eds. 334. Doré I. in The Seafood Industry. or presence of toxin 0. New York. Van Nostrand Reinhold. SMOKED AND DRIED FISH TABLE 5. Horner W.7 1. Van Nostrand Reinhold. 1979.F. 381-406. References Regenstein J. in Fish Processing Technology.A. 56. Urner Barry Publications Inc. Fernandes R. 3273. 1984. in Micro-Facts: The Working Companion for Food Microbiologists. Torry Advisory Note No. London.W. 9. 117 . parahaemolyticus Vibrio vulnificus C. 2007. 1991. monocytogenes Salmonella spp.M. * Staph. Catching. FAO Fisheries Technical Paper. Introduction to Fish Technology. 7. Leatherhead Publishing. Bannerman A. Handling and Processing Eels.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. Wareing P. 1994. Eds. except in the viscera of dungeness crab. 8.8 ppm (saxitoxin equivalent) 20 ppm domoic acid. Flick G. Leatherhead. Assurance of Seafood Quality.M. Martin R.E. Birnie K. Regenstein C... McLay R.. 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. coli (ETEC) L. where 30 ppm is permitted Paralytic shellfish poisoning* Amnesic shellfish poisoning * *These criteria are applied to all fish 5. 37(revised). Marinades. salting and smoking).CURED. 4. Preservation of fish by curing (drying. Hot Smoking of Fish. Food spoilage fungi. Smoked. Horne J. 2. Flick G. Torry Advisory Note No.. botulinum * Limit of tolerance 1x 103 ETEC/g.F. 120-38. 6. The Smoked and Cured Seafood Guide.. 254-82 5. cured and dried fish. USA.. 1990. Fernandes R. Ed. aureus * Vibrio cholerae V. Huss H. 1993. Hall G. Chapman & Hall. 3.
Bioscience. 18. Biotechnology and Biochemistry. 335-40. Food Control.. 118 . Pelroy G. Degradation and omethylation of phenols among volatile flavour components of dried bonito (Katsuobushi) by Aspergillus species..M.. Druschitz A. Shuto Y. Kinoshita Y.E. 51-81. 1995. Martin A. 2324-5. Yamauchi H. 16. Martin A. Bannerman A. 958-60. 59 (12). Eklund M. Food Product Design. Lindemann B. Ninomiya Y. 27 (9).. 1051-5. Bannerman A.C. Teeny F. 20. Bioscience. Ninomiya M. Cann D. 18.K. Paranjpye R. Shuto Y. 13. 56 (6). Torry Advisory Note No. Hosokawa M.. 23. Recommendations for the Preparation of Smoked Salmon. Kinoshita Y. Yokayama S. Torry Advisory Note No. 14. 6-dimethoxy-4-methylphenol by Aspergillus glaucus and their possible contribution to Katsuobushi flavour.R. Biotechnology and Biochemistry. 1386-7... 1981. Journal of Food Protection. Jeppesen V.. Food Chemistry. 1990. Doi M. 19.. 1984. 12.. 2008.... 22 (revised). Cann D. Ogiwara Y. 6 (4).. Microbiological control for fish smoking operations. 61 (8). Torry Advisory Note No. 22. Shuto Y. 5 (revised). Biotechnological Applications. Horne J. 24... Fish Drying and Smoking: Production and Quality. 2007.M. Peterson M.. Oxidation of acetophenone by Aspergillus species and their possible contribution to Katsuobushi flavour. Food Reviews International. Huss H.. Inhibition of Clostridium botulinum types A and E toxin production by liquid smoke and NaCl in hot-process smoke-flavoured fish. Doi M. 21.F.A. Dillon R. O-methylation of 2. 508. 1977. 2002. 17. Ed.. Torry Advisory Note No. 1998. 840-6.. and Biochemistry.H. Technomic Publishers. London.. Chapman & Hall. 17 (9). Kanayama T.C. 45 (10). 1995. Doi M. Bioscience.. Perusing Asian soup options. 11. 6 (6).P. Control of biological hazards in cold smoked salmon production. 108 (3).FISH AND SEAFOOD 10. Flavour constituents of dried bonito (Katsuobushi).E. 1982. in Fisheries Processing. 843. Doe P. Biotechnology. 1992. 53 (4). Embarek P. Hayashi K. Sakakibara H.. Yajima I. Processing Cod Roes. Matsui M. 1997. 15. Matsui M. 1994. Mitou M. Packing Fish in a Modified Atmosphere. 553-72. 88. 935-41. Doi M.B. Aoshima H. Botulism and Fishery Products.W. Shiemori Y. 1980. Lancaster. Agricultural and Biological Chemistry. What’s in a name? Are MSG and umami the same? Chemical Senses. 1989.... Patel T. 25. Halpern B.. Asymmetric reduction of acetophenone by Aspergillus species and their possible contribution to Katsuobushi flavor..M. The discovery of umami. Effect of dried bonito (Katauobushi) and some of its components on GABA-A receptors.
49 (1). 3 (4). Blackie. Hess E... 1979. Gill T. Luxembourg. Food Technology. New York. 181-8. Hocking A.V. Institute of Medicine. Anggawati A. Fungi and Food Spoilage..A. 36. 30. Press Inc. Studies on salt fish. 74. 34. 123-9. 1996.D.I. 1991. Scientific Committee for Food. Seafood Safety. 12.A. V.D.K. CRC. 1994.. Effects of sodium chloride and relative humidity on growth and sporulation of moulds isolated from cured fish.T. Ed.F. US Food and Drug Administration. ceviche. Cox J. Modern Food Microbiology. 5.. 98100. Jay J. Pitt J. Committee on Evaluation of the Safety of Fishery Products.. 42.A.M. Rockville. 164-9. Cann D. 35....CURED. in Smoke in Food Processing. The control of botulism in hot-smoked trout and mackerel..D. 119 . 27. 28. Washington DC. 32. Loessner M. in Fish and Fishery Products Hazards and Controls Guide. 31.. Golden D.1988. SMOKED AND DRIED FISH 26.I.D.. Maga J. 74. Food Microbiology. Lucas F. 1994. 1993. 351-7.H. Ahmed F. Commission of the European Communities. 1961.E. Wheeler K. Diyaolu S. Shewan J.Y. 38 (3)... CEC. 37. Journal of the Fisheries Research Board of Canada. Journal of Food Technology. 218-25. Nahrung.O. Academic Press.. 74 (2). 107-11. Hocking A.A. Journal of Applied Bacteriology. 2005... Incidence of Listeria monocytogenes in an acidified product. Townsend J. Fuchs R. Boca Raton.M. Ed. Smoked and smoke-flavoured fishery products. London. Borgstrøm G. in Fish as Food. An outbreak of Shigella. 33. 14. Røntved S. Lancet. Hansen L. 27686.M. Tour highlights production and uses of smoke-based flavours. Pszczola D. Sprouse R. Antimicrobial properties of smoke. New species of fungi from Indonesian dried fish..E.B. National Academic Press. Letters in Applied Microbiology. Simini B. Food Research International. 39. 22. 70-2. Hocking A. 1971. Pitt J. 1986. 38. Smoke flavourings. Oxford. Pitt J. Adebajo L. The microbiology of sea-water fish.C. Sirvas S. 1991. in Reports of the Scientific Committee for Food (34th Series). 43. Studies on Sporendonema epizoum from ‘dun’ salt fish. 29. Food and Drug Administration. Springer Science. 1996. Food and Drug Administration. Frank M. Fungal flora of South Vietnamese fish and rice. Mycotaxon. 40. Ed. 41.. Importance of autolysis and microbiological activity on quality of cold-smoked salmon. 1997. Interactions among xerophilic fungi associated with dried salted fish.S. Fungi associated with Indonesian dried fish. 1985. 348. Journal of Tropical Medicine and Hygiene.I. Hocking A. Wheeler K.R. Ed.J.F. 29 (2). 311-7. Huss H. 1941. 1995. 1995. 88-90. 742. 197-208. Taylor L.
.W.FISH AND SEAFOOD 44. Clostridium botulinum and its importance in fishery products. Canadian Journal of Microbiology. Cann D.. Round Worms in Fish.. 507-10. 47. 80. Paranjpye R. Appendix 5: FDA and EPA Guidance Levels... Keeping quality of vacuum packed smoked sardine fillets: microbiological aspects. Eklow A. Ribeiro C.C. de Louvois J. Pathogenic microorganisms of importance in seafood...N. 833-41. in Quality Assurance in the Fish Industry. Loncarevic S. 35 (11) 2904-7..D. 1994.html Gilbert R.. Loncarevic S. Roberts D. 120 . Danielsson-Tham M-L. Hurst A. Ericsson H. 3. Amsterdam. 13-8. US FDA Centre for Food Safety & Applied Nutrition. Food Control. Torry Advisory Note No. Fletcher R. 37 (1). 1991.. 1996.. Donovan T. Zeitschrift für Lebensmittel-Untersuchung und –Forschung. Acta Veterinaria Scandinavia. 1992. Hobbs G. Unnerstad H. Pelaez C. Wooten R. Growth and production of toxin of Clostridium botulinum type E in rainbow trout under various storage conditions. 197-209. An outbreak of listeriosis suspected to have been caused by rainbow trout. Peterson M.. Inhibition of Clostridium botulinum types A and E toxin formation by sodium nitrite and sodium chloride in hot-process (smoked) salmon. 55 (4). Eklund M. Journal of Food Protection.. 45 (9).W.8 Further Reading Gibson D. Harrison M. 1980. Little C. in smoked and "gravad" fish. Tham W. 58 (8). Communicable Disease and Public Health. Beltran A. and smoked fish establishments good manufacturing practices. 1976. Association of Food and Drug Officials. Gibbs P... Hughes A. 2000. 5-7.R.H. Susuki E. Tham W. Eds. Cured..A. Bolton F. http://www.. 2001. 863-6. Mentzing L-O. 22. Advances in Food Research. Washington D..L. Liston J. US FDA Centre for Food Safety & Applied Nutrition.. Persson I.V..E. salted.. 5 (1). Journal of the Association of Food and Drug Officials. Ed.. Journal of Clinical Microbiology.S. Richards J. 46. 5. 1980.. 45.M.A. Huang Y. 49-59. Journal of Food Protection.. 232-6.J. 1997. Jacobsen M. Incidence and growth of psychrotrophic Clostridium botulinum in food.J. 26 (4). Prevalence of Listeria monocytogenes and other Listeria spp.fda.. Moral A.. Danielsson-Tham M. Food and Drug Administration..gov/~comm/haccp4x5. 1982. Nye K.C.. Pelroy G. in Fish and Fishery Products Hazards and Controls Guide.cfsan. 1995. Davies A. Huss H.. 188 (3). Garren D. Microbiological guidelines for some ready-to-eat foods sampled at the point of sale.. 1989. 135-85.A. 48.M. Elsevier Science Publishers B. 163-7. The effect of NaCl on the upper temperature limit for growth and enterotoxin synthesis by Staphylococcus aureus.
.R. Dillon R. 21 (3). SMOKED AND DRIED FISH Sutherland J. 1988. 1992. temperature.J.R.. 4389-95. 19..W. International Journal of Food Microbiology. Hitchins A. Efficacy of high sodium chloride concentrations for the destruction of Listeria monocytogenes.. van Spreckens K. 293301.. Applied Microbiology. 11.. 55 (11). Letters in Applied Microbiology. Patel T. Eklund M.. 844-6. 58 (5).. Marine toxins of microbial origin. in a salmon slaughterhouse and smoked salmon processing plant.. 217-36. Incidence of Listeria in hot and cold-smoked fish. 27 (6). Rørvik L. Food Microbiology. Yndestad M. 94-8. International Journal of Food Microbiology.. 1995. Slade A. Prevalence of Listeria in smoked fish. Peterson M. Caugant D. Presence. Beuchat L.N. 121 . 1734.T.. Histamine production by psychrophilic flora.. Davies A. Hudson J.C. Letters in Applied Microbiology. Tran T.S. Liston J. Journal of Food Safety. Fuchs R. Paranjpye R. detection and growth of Listeria monocytogenes in seafoods: a review..CURED. 14.. 11.178-80.. Nicolaides L. Competition of food bacteria with Listeria monocytogenes during enrichment culture. Roberts T. Letters in Applied Microbiology. and salt concentration on growth and survival of Vibrio parahaemolyticus. 1994. 1987.K. Ben Embarek P. Taylor S. 23 (1). Poysky F. 1992. 1075-80. Dallas H. 1994. 1995.A. Combined effects of water activity. Keusch A. 25 (5).A. Fate of Aeromonas and Yersinia on modified-atmosphere packaged (MAP) cod and trout. Bayliss A. Food Technology. Journal of Food Protection. 1994. Ratnam S. in Seafood Quality Determination.A. Elsevier. Predictive modelling of growth of Staphylococcus aureus: the effects of temperature. Contamination pattern of Listeria monocytogenes and other Listeria spp. 866-70... 25.E. 42.. 1973. Pelroy G. Occurrence of Listeria monocytogenes in freshwater farms and fishsmoking plants. Beuchat L.R. 394-6. New York. Jemmi T. International Journal of Food Microbiology. 1995. Eds. 502-8. 1994. Kramer D.P. 309-18. Lashbrook L. 309-16. 19-27. 61 (12). 1974. Applied Microbiology. 1991. Poindexter C. Journal of Food Protection. pH and sodium chloride. and temperature on the growth of Vibrio parahaemolyticus. Incidence and sources of Listeria monocytogenes in cold-smoked fishery products and processing plants. solute. Interacting effects of pH.
It is produced from the sandfish Arctoscopus japonicus and has the rice-based enzyme preparation. Sugar can also be added to the Indonesian fish sauce bakasang and the Thai product nam-budu. FERMENTED FISH Prof. Martin Adams Faculty of Health and Medical Sciences University of Surrey Guildford Surrey GU2 7XH United Kingdom 6. As with most traditional products that are still produced principally on a cottage industry or domestic scale. whereas in those where a carbohydrate source such as rice or sugar is also added. North American and other markets and are.25% and are used mainly as a condiment. though some are produced elsewhere. Essentially.1 Definitions As applied to fish products. they can be divided into two categories: fish/salt products and fish/salt/carbohydrate products.6. the most important of all fermented fish products. The fish sauces are produced on the largest scale. in economic terms. 6. the best-known of the fish sauces (gyoshoyu) of Japan is shottsuru. producing the fish sauces and pastes. they are exported to European. For example. Carbohydrates are added to some fish sauces during their preparation. The relative importance of these two activities depends on the formulation of the product. The principal fish sauces and pastes of south-east Asia and their local names are listed in Table 6.1 Fish/salt products such as the fish pastes and sauces tend to contain relatively high levels of salt.1. typically in the range 15 .I. 123 . there are numerous variants of some common themes and a host of local names used to describe them. koji. lactic fermentation can play a significant role. to those processes in which the activity of lactic acid bacteria (LAB) plays an important part. Fermented fish products are largely confined to east and south-east Asia. the term ‘fermented’ describes a spectrum of processes that ranges from the largely autolytic degradation of fish protein. Those to which only salt is added tend to be mainly autolytic. added during preparation.
This is well exemplified by some of the fish/salt products of Thailand described in Table 6. cheese-like aroma Burma Indonesia Ngapi ketjap-ikan Paste Red/brown salty paste nga-ngapi trassi-ikan trassi-udang (shrimps) prahoc mam-ruoc (shrimps) Kampuchea nuoc-mam nuoc-mam-gau-ca (livers only) Korea myulchljeot jeots. to products analogous to other lactic fermented foods such as salami.FISH AND SEAFOOD TABLE 6. salty taste. a variety of products can be produced within a single country. A selection of Asian products is presented in Table 6. Fermented fish products are of very minor importance outside Asia. In Europe. where the bacterial production of lactic acid is a major feature. 124 . who had adopted them from the Greeks.2 Fish/salt/carbohydrate products range from those that resemble the fish sauces and pastes in the sense that extensive autolysis has occurred. although garos. and pissala.I Fish sauces and pastes of south-east Asia Country Sauce Amber/brown liquid. made from fish livers in Greece.III. These have long since fallen from use. made from whole fish in France. fish sauces known as liquamen or garum were ubiquitous condiments for the Romans.1.II. jeotkals (shrimp) nam-pla (pa) Budu Patis nam-pla nuoc-mam padec Belachan (shrimps) bagoong kapi mam-ca man-tom (shrimps) Laos Malaysia Philippines Thailand Vietnam Even within the limitations of fish and salt as the sole ingredients. may be direct descendants. and the more widely known Worcestershire sauce a very distant relative. 6.
12 months 10% raw sugar added. funa-suchi. burong-ayungi.3 months Main dish Tai-pla Bowels 1:3 6 . mam-seeing sikhae som-kay-pa-eun.g. som-fak/ som-fug 125 Malaysia Philippines Thailand . crustaceans Variety of fish Variety of brackish or marine fish 1:4 3 . shrimps. pa-khem. som-pa.II Fermented fish/salt products common in Thailand Name Type of fish Form Approx. ayu-sushi. mam-chao.4 months Condiment and main dish Whole 1:3 18 months Condiment Whole 1:3 After 3 .8 months Main dish TABLE 6.FERMENTED FISH TABLE 6. the mixture is then boiled and bottled - Condiment Nam-khoei Shrimps. pla-chao. tai-suchi phaak. e. pa-chao. som-pa-keng pekasam.g. cencalok burong-isda. crustaceans Mackerel eviscerated fish Variety of fish Whole 1:4 Condiment Pla-thu-khem Whole 1:3 2 . pla-som. mam-pa-kor. burong-bangus pla-ra. e. burong-dalag.III Fish/salt/carbohydrate products of south-east and east Asia Country Japan Kampuchea Korea Laos Products I-sushi. 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.
These are often described as fish/salt products. especially in fish/salt products. which employ lower levels of salt than Asian products. malt or flour (1). in Norway and surstrømming. Table 6. Penaeus indicua Many of the fish used in fermentation are uneviscerated. A modern account of their production. The diversity of fish used in fermented products (see. Here. For example. 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 the initial microflora is likely to be considerably larger in these cases. rakefisk or surfisk. states that sugar is often included with the salt. TABLE 6.IV) and in the way in which they are handled prior to processing means that the initial microbial levels are far from uniform. and their microflora tends to reflect their local environment more than that of marine species.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. Norwegian gravlaks. it was found that several different sources were used for the fish . and a large local freshwater reservoir (2). and counts ranging from below 104 /g to in excess of 107 have been reported. in a survey of fermented fish products in north-east Thailand. however. fermented trout. 126 .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. which has been increasing in popularity over the last 50 years.the flooded rice fields. This could lead to quite marked variations since the same fish can be obtained from a variety of sources. or buried salmon. some fermented fish products remain popular. There are also more heavily fermented products. the most popular varieties of which are rakørret. for example. paddy ponds beside rice fields used for collecting fish when the field has dried up. made from herring. 6. relatively mild-tasting product.FISH AND SEAFOOD In Northern Europe. is a traditional. and old descriptions of their production often refer to the addition of whey. in Sweden. freshwater fish are the usual raw material.
In some cases. This.3. however. Similarly microbial inactivation by irradiation has not been found to adversely affect the properties of Korean fermented squid (7). These preparations contain a mixed microflora of moulds and yeasts. and the anaerobic conditions that prevail in the bulk of the material. either directly or as the partially saccharified rice they produce. Hansenula. In another microbiological study of four fish sauces. after one month. Other halophilic species have been identified from fish/salt products. were found to be the predominant isolates.3 6. 6. angkak (Philippines and Thailand) and look-pang (Thailand) are used. 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. 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. Bacillus spp. in many cases. in numbers reaching 108 /ml after 3 weeks of fermentation but declining thereafter. they can contribute to the initial microflora. however. principally species of the genera Rhizopus.organisms that play an important part in the production process but can also contribute to the ultimate spoilage of the product. When this is a simple carbohydrate source such as sugar. More recently. Endomycopsis and Saccharomyces . sufficient to saturate the water phase present and produce an aw of 0.4. where the total counts dropped from above 107 cfu/g to below 103 cfu/g after 14 days and below 102 after 40 days. It also demonstrated their significant proteolytic activity during the first month of fermentation. Studies have shown that the number of bacteria declines rapidly during the production of the Thai fish sauce nam pla and that.FERMENTED FISH Where other ingredients are used. suggesting that halophilic bacteria do play an important role in the production of fish sauce (8). which included nam-pla and patis. work with nam-pla in Thailand has isolated Halobacterium and Halococcus spp. the product contained about 500 cfu/g comprising mainly Micrococcus and Bacillus spp. the Philippine fish sauce. and studies 127 . Similar results were seen with patis.74 or below. mean that microorganisms have been generally held to play little or no role in the process that follows. (3. It has also been shown that a perfectly acceptable patis could be produced from fish that had been sterilised with ionising radiation (6). Generally. probably reflecting their ability as spore-formers to survive rather than any capacity to multiply under the prevailing conditions (5). or boiled or roasted rice.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. 4). Mucor. its contribution is likely to be insignificant compared with that of the fish microflora in terms of both spoilage potential and public health concerns. traditional starter culture preparations such as koji (Japan).1 – 6.
6. 6.FISH AND SEAFOOD have demonstrated the production of halophilic proteinases from fish sauce isolates (9 .1.12). Patis (Condiment) 128 . Fig. Belacan (Condiment) Fig.2.
thought to be largely the result of autolytic breakdown of the fish tissues. however.3. 14). 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. To produce semi-solid fish pastes as opposed to fish sauces. and muscle cathepsins have been shown to be inhibited at relatively low salt concentrations (13.4. Myulchijeot (Staple food) The extensive proteolysis and liquefaction that occur during production are. the 129 . 6. In fish sauce production.FERMENTED FISH Fig. Ika-Shiokara (side dish) Fig.6.
V and Figures 6. Some examples of products and their formulation are presented in Table 6. 6.5). Their main feature is that the provision of a source of carbohydrate allows a lactic acid bacterial fermentation to occur.3. Figure 6. and the low pH produced contributes a second protective barrier or hurdle against the growth/survival of the normal fish-spoilage organisms and pathogens. particularly glutamate. Hoi-Malangpu-Dong (side dish.8. volatile fatty acids and nucleotides. In contrast to free-swimming fish. for example.2 Fish/salt/carbohydrate products of Asia This category encompasses an even greater diversity of products than the fish sauces and pastes. filtered and bottled.FISH AND SEAFOOD high salt level used also assists in the osmotic extraction of a supernatant rich in amino acids. Fig.5.6 – 6. snack) 130 . shellfish such as mussels often have appreciable levels of carbohydrate in their tissues and in these cases addition of carbohydrate is not necessary for a lactic fermentation to occur (see. which is eventually separated from the residual fish tissues. 6.
12 days 3 .5-1.12 months Shelf life Use 1 .6. C.12 months Main dish 2 Pla-jao C. 3:1:0. north.10 days 5 days Main dish Main dish Main dish Condiment and main dish Note: Products 1 .FERMENTED FISH TABLE 6. 3:1:1-3 Fish:salt:boiled rice: garlic 10:2:1:0. Kung-Chom (staple food. N S 5 . north-east. south of Thailand.20 days 3 Pla-som 4 Pla-jom 5 Som-fak 6 Pla-paengdaeng NE. C NE.2-4 Fish (pieces): salt:khaomark (fermented rice). Fig.4 are also produced using shrimps.5:2-3:1 Fish:salt:boiled rice: ang-kak 3:1:3:0.25-1 Fish:salt:roasted rice: garlic 10:1:3:1 Fish (minced):salt:rice: garlic 10:0. S. the prefix 'pla' is replaced by 'kung'. NE. central. N. NE 10 .03 Preparation time 6 . Ang-kak: red mould rice obtained by fermenting rice with Monascus purpureus. NE Formulation Fish (whole or pieces): salt: roasted rice.7 days 5 . side dish) 131 . N.V Fish/salt carbohydrate products of Thailand Name 1 Pla-ra Region C.6.3 years Condiment and main dish 2-3 months 3 weeks 2 weeks 2 weeks 6 . N. C NE. In these cases.
1 The use of higher salt levels results in a longer production phase but a better keeping quality product.VI). 106) Fig. (from Arroyo et al. If acidity is a desired sensory property of the product.. then this will take longer to develop than if lower salt levels are used.7. the salt is of far greater importance in determining the product’s long-term stability.FISH AND SEAFOOD The extent to which lactic fermentation occurs depends largely on the salt content. The LAB isolated from these products are those with more marked salt tolerance but. 2. these products are salted at lower levels than the fish sauces and pastes. typically in the range 2 . 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 6.6. This is readily explained by the susceptibility of LAB to elevated salt concentrations.3. A survey of fish/salt/carbohydrate products in Thailand (15) noted that there were two empirical rules governing formulation:6.2. their growth and metabolism are nonetheless slowed. Generally. but there is considerable variation in this. of the two factors contributing to shelf life . while they are able to grow at high salt levels. Red burong isda 132 . It also indicates that. giving a reduced rate of acid generation.10%.salt and lactic acid. Phillipines Journal of Food Science and Technology.
FERMENTED FISH (from Arroyo et al. 106) Fig. 2.4 .1 .9 4.8 pH 4.4. 133 .5.4. White balao-balao TABLE 6.0 .12 months 1 .6.VI Shelf lives reported for fish salt carbohydrate products Product Pla-som Pla-paeng-daeng Pla-ra Pla-chao Pla-chom Som-fak Salt 2.2 7.0 .7 .3 years 1 .6.3 .8 .6 3.5 .8 .5 .3 5.9.9 .8 2.5.9 126.96.36.199..0 Shelf life 3 weeks 6 .2 years 2 weeks 2 weeks Source: A Concise Handbook of Indigenous Fermented Foods in the ASCA Countries (16). Philippines Journal of Food Science and Technology.0 .9.2 188.8.131.52 4.5 3.1 4.6.
TABLE 6. also serves as a valuable source of the more readily fermented carbohydrate.4. efficient lactic acid production will depend upon the presence of amylolytic LAB or saccharification by other microorganisms or enzymes that may be present. When added as boiled or roasted rice alone. they increase the amount of soluble sugars produced and thus the range of LAB that can grow. 21). such as salt content. Garlic. inulin.9) compared with the white product (pH 4. Addition of carbohydrate will. although the final population of LAB may be similar. a saccharifying agent such as koji (Japan) or ang-kak (the Philippines) may be added separately (Table 6. Although the ability to ferment starch is not widespread in the LAB.5) despite other factors. For example. the rice may be added in a partially saccharified form such as kao-mark (Thailand). 19). Where they are used.VII). alternatively.VII Fish/salt/carbohydate products in which saccharified rice or a saccharifying starter culture is used Product red burong-isda i-sushi.3. Rice is the most common carbohydrate source used.0 . This results in a more active lactic fermentation and a lower pH.1 . 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 . a common ingredient in these products. being similar (18. In several products.2 Inclusion of more carbohydrate gives a faster fermentation and a stronger acid taste Increasing the proportion of carbohydrate in the formulation decreases the product’s buffering capacity and provides the LAB with more substrate on which to act. 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. also lower the unit cost of the product and this is probably a significant factor in many cases. a survey of burong-isda in the Philippines reported consistently lower pH values for red burong-isda where ang-kak was used (pH 3. of course.FISH AND SEAFOOD 6.3. 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). The effect of this will be to decrease the product’s final pH and increase the amount of total acidity.
The growth of yeasts and moulds could.7 up to 6. such as wrapping in banana leaves. Results of a product survey were consistent with this. It seems that a lactic fermentation takes place in many types of pla-ra but is not significant in others. The mixture is allowed to ferment for 2 . Excessive growth and acid production by the LAB could render products too sour for acceptability. Clearly. 135 .2 at the end of fermentation. which is sufficient to prevent lactic fermentation by all but the most halotolerant LAB. Cleaned pieces of eviscerated fish flesh are salted with 6% salt overnight before mixing with cooked millet. 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. is something of an exception since millet is used along with garlic as the carbohydrate source.9%.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). which ranged from 7. markedly extending the shelf life (25). will not reliably exclude air. A combination of irradiation and chill storage has been shown to prevent both over-acidification and fungal growth in som-fak.8 . allowing other spoilage organisms to grow. Any loss of acceptability must arise from chemical or physical changes occurring over prolonged storage. thereafter declining to about 4. There is little published information on the factors that limit the shelf life of the low-salt products.5 in the first 2 . reporting the product’s pH to range from 4.5:9:3. 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). In a successful fermentation.5. and there is a marked regional variation in the preferred type.FERMENTED FISH Sikhae. Generally. the level of salt employed is on the high side for fish/salt/carbohydrate products. One study found that any effective measure to exclude air from a fermenting fish/carbohydrate mix prevented yeast growth (23).3 weeks at around 20 °C.7 to below 4. Traditional packing techniques used with lactic fermented fish products. lead to an increase in pH. This suggests that use of oxygen-impermeable wrapping films could extend the shelf life of such products. but in most cases it is probable that yeast and mould growth would be an important cause of spoilage. in turn.3 days. the pH drops from 6. but they are likely to behave similarly to lactic fermented products produced from other commodities. red pepper powder and garlic in the ratio fish:millet:pepper:garlic of 80:7. The Thai product pla-ra exemplifies these basic rules as well as the difficulties associated with attempts to classify such products. presumably correlating with the salt content. and surface mould growth has been shown to be a cause of rejection in the Thai product som-fak (24). 6. the salt levels in the latter are sufficient to rule out microbiological spoilage in all but the most exceptional of cases. the Korean fermented fish product.17. around 12%.2.
There is little in the way of documented information associating fermented fish products with foodborne illness. After capture. The combinations of pH and salt in many fermented products are likely to preclude growth of the organism completely and. and consumed without cooking. parahaemolyticus has been reported growing down to a pH of 4. viable numbers will probably decline quite rapidly. V. Because of the severity of the illness it causes. In the fish sauces and pastes.5 Pathogens: Growth and Survival Many fermented fish products are stored at ambient temperatures. typically 30 °C. but we are unaware of any reports of this happening. The fish/salt/carbohydrate products depend on the combination of salt and pH/lactic acid for their safety. As in Norway. however. Their safety therefore is critically dependent on the quality of the raw materials used and the inhibitory effects of the fermentation process. under conditions inimical to growth and high ambient storage temperatures. 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. are Vibrio parahaemolyticus and Clostridium botulinum. 6.8 and at salt levels as high 10%.87. the pH decreases below 5 and safe processing is recognised to depend on proper salt levels and temperature control until the pH drops. The epidemiological evidence points to greater awareness of the importance of these factors since outbreaks have declined over recent years (27). and failure to achieve a sufficient combination of inhibitory factors could lead to their growth or survival. fish products are the most common source of botulism. During fermentation. The bacterial pathogens most commonly associated with the living environment of fish. outbreaks of 136 . fish can potentially be contaminated with almost any foodborne pathogen as a result of improper handling and storage. which accounted for 88% of outbreaks in the period 1952 . 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.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. and the most frequently implicated product is the fish-salt-carbohydrate fermented product Izushi. Improper storage of fish prior to processing could result in the production of toxins capable of persisting through to the product. Several measures have been suggested for reducing the risk of botulism. In Japan. The Scandinavian fermented fish product rakefisk was responsible for 16 (84%) of the outbreaks of botulism recorded in Norway in the period 1961 90. although these limits apply when all other factors are optimal (26).
1966. References Riddervold A. 3. Salting levels are often insufficiently low to prevent growth of C. beaver tail and paw. while increasing the acetic acid content of the formulation had little or no effect (32). Liston J. Lutefisk.. Mahidol University. whale meat. Saisithi P. giving an alkaline fermentation. An investigation of survival of the fish parasitic nematode Anisakis in marinated herring has given some indication of what these studies might find. Raw materials subject to this process include seal flipper. they could act as an important vehicle of transmission. Rakefisk and Herring in the Norwegian Tradition. Yhoung-Aree J. 105-10. and these are often used in the production of fermented fish products. Foodborne trematode infections are emerging as a major public health problem. Journal of Food Science. 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). although the absence of any source of fermentable carbohydrate means that the process is putrefactive. Baseline Study (1987-1988)... Many of these products are described as fermented. salt content was found to have the greatest impact on the viability of larvae. A case-control study conducted in northeast Thailand showed that elevated levels of antibodies against O. Novus Press. particularly in south-east Asia and the western Pacific regions (30). Fermented Fish Products Study (Thailand) Phase 1. Thailand. 1990. 31. One study with the trematode Haplorchis taichui. and warm weather can result in rapid growth of C. walrus nose.6 1. Institute of Nutrition. showed that inclusion of fermented fish in the Thai dish lab-pla improved inactivation of the metacercariae. Freshwater fish are the most important intermediate reservoir hosts of foodborne trematodes. With this particular organism. 1989. probably reflecting greater awareness of the critical factors that determine the product’s safety (27). The more widespread current use of non-traditional containers such as glass or plastic jars is thought to be one contributing factor (28). Infection with the trematode Opisthorchis viverrini is associated with cholangiocarcinoma. Botulism has also been associated with a number of traditional foods of the peoples of northern Canada.. Kasemsarn B. though 27% survival was still recorded (33). 6. Puwastien P. botulinum. Dhanamitta S. 2. which will persist into the product. They are packed into containers such as buckets or pits in the ground and left at ambient temperature to putrefy. 137 .O. Oslo. affecting an estimated 40 million people. Microbiology and chemistry of fermented fish. viverrini were a strong risk indicator and that consumption of fermented fish products was an independent risk factor in the contraction of cholangiocarcinoma (31).FERMENTED FISH botulism have become less frequent in recent years. botulinum and the production of toxin.M. Dollar A. Where these products are consumed without final cooking. Greenland and Alaska. which is generally consumed raw (29).. and salmon heads and eggs.
5. 11. Chung Y. 1995.. 8. 9. 63... International Journal of Systematic and Evolutionary Microbiology.. Backhoff P. Yongsawatdigul J. New York. 138 . 106-8.M.. Process Biochemistry. Journal of Food Science. 80. in General and Applied Aspects of Halophilic Microorganisms. Yasir M. Sands A. Reddi P.H.. 1972. 185-92. 29. Oda K.. Liston J. 3649. Journal of Food Science and Technology. Kaneko Y.FISH AND SEAFOOD 4. microbiological and chemical properties of salted and fermented squid. 1986.. Journal of Fermentation and Bioengineering.. Itoh T.. Cooke R. Production and characterisation of NaCl-activated proteinases from Virgibacillus spp.T. Saono S. Journal of Scientific and Industrial Research (India). Im W.... Journal of Food Protection. Tsunemi M.. Orejana F.. Agents of proteolysis and its inhibition in patis (fish sauce). 15. Lentibacillus kapialis spp..H..R. 1395-401. Phil.. A Concise Handbook of Indigenous Fermented Foods in the ASCA Countries. Canberra..K. 25. Namwong S. 43. Dong-Ho K.R. 1975. Kyong-Haeng L.W. SK33 isolated from fish sauce fermentation. Constantinides S. 1991. 61-73.. Thongthai C.. 11. Bioscience. 353-63. Moonmangmee S... 17-30. Ono H.. 381-8.K.R. a traditional Korean fermented seafood. SR5-3 isolated from fish sauce: purification and characterisation. 1985. 1976. Catheptic activity of fish muscle. Dhamcharee B. Nov... 10. A halophilic serine proteinase from Halobacillus spp. 2007. Some chemical changes in fish silage. 12. Tanasupawat S. Crisan E.. Lee S. Rattagool P.J.. Velankar N. 57. Thawai C. 124-30. 2006. 6.. from fermented shrimp paste in Thailand. 643-7.. 16. Rodtong S. Government of Australia. Yook H. Rodriguez-Valera F. Tanasupawat S. Ed. Olympia M. Takada K. Halophiles in Thai fish sauce (Nam pla). Dymsza H. Suntinanalert P.T. 7. 6338. 13. Tropical Science.A. Byun M. Applied Microbiology. Journal of Food Technology.M... Biotechnology and Biochemistry. 14. Effects of gamma radiation on sensory qualities. 2000. International Journal of Systematic and Evolutionary Microbiology. Hull R. 2007. 1957. Nov.. Kudo T. 1979. Jae-Hun K. 141-2. Adams M.V. isolated from jeotgal. 2008. Fukuda H. 17. 934-9.S. Hiraga K. Fermented fish products of south east Asia. 16A. 57. 70. 3. Sinsuwan S.. Plenum Press... Characterisation of starch-hydrolysing bacteria isolated from a fermented fish and rice food “burong isda” and its amylolytic enzyme.D. Microflora of four fermented fish sauces. 37. Pakdeeto A.. Aslam Z. Lim J. Ahn H. Salinococcus jeotgali spp. Protein hydrolysate from fish. Takano M.
in Clostridium botulinum: Ecology and Control in Foods.H. The fermented fish food...H. Patrick D. Hazards from northern native foods. Valyasevi R.W. 1996.A. Kozaki M. Eds... 92. Hauschild A. Food Chemistry. 69-104. Genotypic and phenotypic characterisation of garlic-fermenting lactic acid bacteria isolated from som-fak. 27. Ostergaard A.K. International Commission on Microbiological Specifications for Foods. Avurhi J.. Isaac-Renton J. ASEAN Food Journal. Wainwright R. a Thai low-salt fermented fish product.. burong-isda. International Commission on Microbiological Specifications for Foods. Gram L. Sakai H. Marcel Dekker. Journal of Agricultural Science (Tokyo).L. Kozaki M.. Moody L.. Dawar M. 274-86. Huss H... Martin J. 28. Ed.. Dodds K. Vibrio parahaemolyticus. Hauschild A. 1-4.. 20. Rustad T. New York. Dodds K. 24.L. Effect of irradiation on properties and storage stability of Som-fug produced from bigeye snapper.... Gram L. 23. 21. 3. Tanaka M.D.H.. Huss H.H. Control of foodborne trematode infections. Caldo G. Journal of Applied Microbiology. 139 . 38.. 30. 2007.M. Fermentation and spoilage of som fak.H. Gram L. Hauschild A. Characterisation of lactic acid bacteria isolated from a Thai low-salt fermented fish product and the role of garlic as substrate for fermentation. 2002. International Journal of Food Microbiology..B. Visessanguan W. 1995. Ben Embarek P.H. 22. Journal of Food Science and Technology.A. Erikson U. a Thai low-salt fish product. Riebroy S. London.. Huss H. 103.FERMENTED FISH 18. in the Philippines.W.B. 104-6.. 28. Two outbreaks of botulism associated with fermented salmon roe – British Columbia. 1992. Sakai H. World Health Organisation Technical Report Series. Epidemiology of human foodborne botulism. 19. 307-14.. 1992. Eds. Marcel Dekker.. 28. 28 (6). 138-44. 1999. 3-10. Canada Communicable Disease Report.. 105-12. 1987. The influence of oxygen accessibility on the growth of yeast in fish/rice fermentation. Tropical Science.. Traditional fermented fish products with special reference to Thai products. in Microorganisms in Foods. WHO Study Group. 27. Fung C. 305-22. 1815.. Journal of Agricultural Science (Tokyo). Owens J. 219-29. 2002. Benjakul S. in Clostridium botulinum: Ecology and Control in Foods.W. Blackie.. Paludan-Müller C. Saisithi P. 1983. 1983. Yamprayoon J. 26. 426-35. 25. Caldo G. New York. Paludan-Müller C. 29.D. 1998.. 1990. Volume 5: Microbiological Specifications of Food Pathogens. 46. Yeast-flora in red burong-isda a fermented fish food from the Philippines. Wedell-Neergaard C.
. Todoroki T.FISH AND SEAFOOD 31. 2005. Fermented flesh foods. Southeast Asian Journal of Tropical Medicine and Public Health... 117. Gaithersburg. Horst K. Thai traditional food preparation. Deerasamee S..R. 33.R.. 2005. International Journal of Cancer. 854-60.7 Further Reading Adams M. Bloemsma B.H.. Nithikathkul C. Miwa M. 1986. Fermentation and Food Safety.. Chuboon S. Adams M. Srivatanakul P.. Huss H.. 23. Wongsawad C. 15998. 2001. Honjo S.. 661-70.J. 32. Sriplung H. Kittiwatanachot P. Kikukawa H. International Journal of Food Science & Technology. Progress in Industrial Microbiology. 6. Uchida K.. Nout M. Sripa B. Survival of Anisakis larvae in marinated herring fillets.... Aspen Publishers Inc. 36. Roepstorff A. Jedpiyawongse A. Survival of Haplorchis taichui metacercariae in lab-pla. 1995. 29.. Hanai S. Ruamsuk A. 110-1.. northeast Thailand. 140 .. Genetic and environmental determinants of risk for cholangiosarcoma via Opisthorchis viverrini in a densely infested area in Nakhon Phanom..R.
While some of these toxins only occur in seafood from a restricted geographical region. most notably the azaspiracids and the cyclic imines as well as some additional polyethers. more traditional methods such as the mouse bioassay continue to be used in many countries to warn of the presence of toxins. Increasingly sophisticated instrumentation (e.7. Since the original chapter in this series was published (6). known and unknown. physiology and detection of marine and freshwater toxins has also appeared (8). Scientific and Cultural Organisation) has been updated (7).M.1 Introduction Shellfish and finfish toxins continue to be a threat to public health and local fisheries around the world. produced by the Intergovernmental Oceanographic Commission (IOC) of UNESCO (United Nations Educational. FCIC UNC Wilmington Center for Marine Science Marvin Moss Lane Wilmington NC 28409 United States of America 7.C.. and a second updated edition dealing with the pharmacology. including the well known marine toxins 141 . It remains to be seen if this approach gains momentum in the future. the export of exotic seafood that may contain such toxins can lead to unexpected public health issues. several scientific reports have described their efficacy and potential. Ph. The manual on harmful marine algae and their products. Liquid Chromatography/ Mass Spectrometry/ Mass Spectrometry -LC/MS/MS) is employed to identify and confirm the presence of known toxins.D. 5).g. Another book dealing with the clinical effects of a variety of poisons and toxins. C. Wright. Although most of these antibody-based test kits have yet to receive international regulatory approval. In the last decade seafood monitoring has intensified and detection methods have continued to develop in selectivity and sensitivity. An emerging technology is the use of immunosensors (1 . a number of new toxins and toxin families have been identified. and is often necessary in light of the complex toxin profiles that can occur in shellfish tissue. Another important development is the introduction of sensitive and selective antibody-based detection methods for a number of toxins. FISH AND SHELLFISH TOXINS Jeffrey L.3) and some practical applications for detecting shellfish toxins have been reported (4.. Nevertheless.
The remarkable frequency of neurotoxins from marine dinoflagellates has been noted (13). Consequently. all of which contain a tetrahydropurine skeleton (Figure 7. with intoxications occurring in many countries even today (15). In addition. and a tropical species Pyrodinium bahamense (16). dealing with the detection methods and regulatory requirements for many marine toxins has been published (12).1 Origins and distribution The PSP toxins are produced by several dinoflagellate species belonging to the genera Alexandrium. 142 . PSPs are watersoluble and heat stable so cooking has little or no effect on PSP toxicity. and another. even crustaceans such as lobsters. 7.FISH AND SEAFOOD has been produced (9). and a review on the risk assessment of marine toxins has also been produced (14). Many of these species are extremely common and are found throughout the world in a variety of cold. Not all the toxins found in shellfish extracts are microalgal products. An account of this meeting detailing recommendations and scientific advice on marine biotoxins has been published (10). temperate.g. Around twenty PSP toxins are known.1).2. and sub-tropical waters and habitats (15). The Ncarbamoyl sulfates are readily hydrolysed to STX. Saxitoxin Group instead of the Paralytic Shellfish Poisons). 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. Gymnodinium catenatum. 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. A brief report on the status of rapid detection methods for phycotoxins in the European Union (EU) has appeared (11). 7. can become contaminated with these toxins. Saxitoxin (STX) and neosaxitoxin (neoSTX) are the parent members of the group and are the most toxic. neoSTX and Gonyautoxins (GTX) 1-4. All PSPs degrade rapidly in alkaline conditions. a great variety of shellfish from all latitudes.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. some are the result of biotransformation processes by the filter-feeding host.
tongue.30 minutes of consumption. this is never sufficient to render the seafood safe to consume. PSPs bind to site 1 of sodium channels reversibly blocking sodium conductance in nerve and muscle membranes by binding to a specific receptor site located on the outside surface of the membrane close to the ion channel opening.1.2. and humans seem to be particularly susceptible. and fingers within 5 . Cooking does not destroy these compounds and although some of the toxins may be extracted into the cooking water. Human symptoms begin with tingling and numbness of the lips. the less potent C toxins are readily hydrolysed by acid to the more potent GTX toxins. 143 .2 Toxicology PSP toxins affect the central nervous system of mammals. However.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. Structure of saxitoxin and neosaxitoxin and their sulphated derivatives 7. leading to paralysis within a few hours and in extreme cases death by asphyxiation due to respiratory paralysis. 7. Neurophysiological studies have shown that PSP toxins all possess the same mode of action though some derivatives such as STX and neoSTX are considerably more potent than the C-toxins (15).
other derivatives have been reported.FISH AND SEAFOOD 7. a criticism has been that such methods are time consuming. 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.2). 7. However it remains as the monitoring method of choice in many countries. and over ten analogs are known (22. After some development and various trials. 24) (Figure 7. a pre-column method has received Association of Analytical Communities (AOAC) approval (18). TTX most commonly occurs in many species of puffer fish. An antibody-based kit that can detect STX in shellfish is commercially available (RIDASCREEN®). Taiwan. and a second rapid dipstick kit (MIST Alert™) is available for the qualitative detection of positive and negative samples (19). 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.3 Detection methods The safe limit set by most countries for PSP toxins is 800 µg/kg (0. There continues to be a significant effort applied to the development of alternative chemical and biochemical detection methods for these toxins. specifically 144 . and while simple and relatively inexpensive to operate. Alternative HPLC methods have subsequently reduced this to a single HPLC run. namely oxidative conversion to fluorescent derivatives after High Performance Liquid Chromatography (HPLC) analysis (post-column derivatisation). and more standards of individual PSP toxins are required to validate the complete profile to toxins that could occur in contaminated shellfish tissue.1 Origins and distribution Unlike many other marine toxins there is no temporal distribution of TTX episodes.3 Tetrodotoxin Tetrodotoxin (TTX) possesses a unique chemical structure and was the first marine toxin to be chemically characterised.2. The mouse bioassay has been in place for decades and the method has been standardised. One approach has been to take advantage of the ready conversion of these toxins to fluorescent derivatives. or conversion before HPLC separation (pre-column derivatisation) and these have recently been thoroughly reviewed (12. 15. Thailand. and islands of the South Pacific. China. This kit was found effective in all the samples tested in Europe (20) and was applied recently in a survey of Scottish shellfish (21). 17). including Japan. as well as other parts. and has also been evaluated by a number of EU laboratories. While the chemical detection methods are most promising for the future. Early post-column methods were somewhat time-consuming and required three separate HPLC runs to complete the analysis.3. 23. Most intoxication events have occurred in South East Asia. Since its discovery. 7. Two approaches have been taken here.8 µg/g) of shellfish or seafood tissue.
Severe poisoning causes paralysis and respiratory failure. raising speculation that TTX is produced by a bacterial source. A review of TTX detection methods has been published (12. it is served in speciality restaurants with qualified chefs to properly prepare the fish.3 Detection methods TTX can be extracted from suspect tissue using dilute acid conditions. and even newts. and blocks diffusion of sodium through the sodium channel. In Japan. Typically this has been accomplished using HPLC followed by post 145 . and more recently by an actinomycete Norcardiopsis dassonvillei (26). However TTX has been found to occur in a remarkable variety of genetically unrelated species including gastropods. and ingestion of larger amounts leads to cardiovascular effects including bradycardia. horseshoe crabs. and in mild cases results in distal muscle weakness. 27).2 mg of purified toxin can be lethal. All of the observed toxicity is secondary to the action potential blockade. This was indeed confirmed and a number of marine Vibrios have been shown to be capable of TTX production (24.2.3. The onset of symptoms usually occurs rapidly. hypotension. nausea and vomiting. this has expanded to include bacteria within the Alteromonas and Pseudomonas genera. 25. and once again the mouse bioassay has been used to detect the presence of the toxin. 7. An oral dose of 1 . Fig. the blue-ringed octopus. 7. Since then.2 Toxicology Like the PSP toxins. TTX acts on the central nervous system by binding extracellularly to site 1 of voltage-gated sodium channels. preventing depolarisation and propagation of action potentials in nerve cells. 26). respiratory failure and coma. 23. gonads and roe (24).3.FISH AND SHELLFISH TOXINS in the liver. molluscs. where puffer fish of the genus Fugu are a delicacy. 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. Structure of tetrodotoxin 7.
4. Immunoassay methods have also been investigated. a polar glutamate agonist. 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).1 Origins and distribution The cosmopolitan diatom Pseudonitzschia multiseries was identified as the culprit organism in the 1987 poisoning event (31). Structure of domoic acid and three naturally occurring photoisomers 7.3 for examples). and while promising. These isomers arise as a result of isomerisation and location of the double bonds in the side chain of the molecule. Surveys by a number of groups have identified over a dozen species of Pseudonitzschia capable of producing DA.FISH AND SEAFOOD column oxidation to the fluorescent derivatives. but all of them are considerably less toxic than DA itself (29). Fig. need further development (24). 7. However this method has been criticised for being complex. Since the original episode of human poisoning following consumption of DA-contaminated shellfish in Canada in 1987 (28).4 Amnesic Shellfish Poisoning (ASP) Toxins The parent compound of this group is domoic acid (DA). but 146 . Currently 12 isomers of DA have been reported (see Figure 7.3. but it is known that such congeners are less toxic than DA itself (30). 7. both the toxin and the diatoms capable of producing it have been found to be distributed worldwide.
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. found in the Pacific around Vietnam. the Philippines. but any impairment of the blood brain barrier results in additional risk.FISH AND SHELLFISH TOXINS only three strains (P. and clams. Penetration of the blood brain barrier is poor in healthy subjects. and review of the toxicologic pathology of DA has been published (36). scallops. 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.10% of the administered dose. nausea. seizures. and Pseudonitzschia seriata) are considered to be high producers of DA (>10 pg/cell). herbivorous fish become toxic after ingesting strains of Pseudonitzschia spp. Recently. 33). To a great extent. and in locations around New Zealand (32. cormorants and pelicans (34). Another factor is the movement of the toxin through the food chain. 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. The oral absorption of DA is around 5 . has been reported as a medium-level producer of DA (33). disorientation and coma. 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. the Mediterranean. Okinawa and more northerly parts of Japan.. 7. including persistent short term memory deficits (36 . another diatom species Nitzschia navisvaringica. Pseudonitzschia australis. Indeed. multiseries. which has been well documented in studies along the west coast of the US. 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.39).4.2 Toxicology DA is an acidic amino acid and potent neurotoxin unaffected by cooking or steaming. The effects of DA may be augmented by the excitotoxic effects of glutamate and aspartate in the sample. 147 . and sea animals such as sea lions and even whales (35). Levels of DA in the other strains are generally <1 pg/cell. as does impaired renal function which increases serum concentration and residence time of the toxin. southern and western portions of Ireland. the wide distribution of toxin-producing strains of the diatom have resulted in the frequent occurrence of DA in many filter feeding organisms such as mussels. For example. Victims display a variety of symptoms including headache. and this has led to ASP toxicity in birds.
and consequently were often overlooked as the real source of the toxins. The sensitivity of the LC/UV (ultra violet) method (detection limit ~ 1 µg/g) can be improved (20 . the term “DSP toxins” will only be used to refer to these three polyether compounds and any derivative based on them. as can LC/MS. and dinophysistoxin-2 (DTX-2) (Figure 7. DTX-2 has been found to be extremely widespread throughout many European maritime countries and is often the major DSP toxin present. California. although in the latter study the probes for Pseudonitzschia pungens. and an immunosensor method has also been reported (4). However. australis. are amenable to laboratory culture and consequently are used in many experiments involving studies of DSP toxins. the heterotrophic dinoflagellates Dinophysis spp. are the more potent source of toxins – often very low cells/ml of seawater can result in human poisoning. dinophysistoxin-1 (DTX-1). and these methods have been reviewed (40. Provided the concentrations in shellfish are high enough (>50 µg/g shellfish tissue). 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). Dinophysis spp. 41). In the context of this chapter. To further complicate matters. Alternatively.4).FISH AND SEAFOOD 7.4. though this method is most commonly used to analyse plankton and seawater (42). 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. DSP toxins are produced by dinoflagellates of the genera Prorocentrum and Dinophysis (47. 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. An EnzymeLinked ImmunoSorbent Assay (ELISA) kit (Biosense) for DA has been approved by the AOAC. 48. produce a variety of other 148 .3 Detection methods DA is water soluble and can be extracted from shellfish tissue by the acidic PSP extraction method although this may result in some decomposition. Since then.30 ng/g) with a rapid clean-up step using a strong anion exchange resin. but just over 15 years ago a third toxin.. Since their first characterisation it was believed that the only causative agents of this group were OA and DTX-1 (45). Of additional interest is the use of molecular probes originally developed to detect the presence of cells of potentially toxic Pseudonitzschia spp.5 Diarrhetic Shellfish Poisoning (DSP) Toxins The active agents responsible for DSP incidents are a group of polyethers that include okadaic acid (OA). The probes have been employed in trial studies in New Zealand (44) and Scotland (43) to detect P. Detection levels can be further lowered by conversion of DA to the fluorescent fluorenylmethoxycarbonyl (FMOC) derivative. Pseudonitzschia fraudulenta and Pseudonitzschia delicatissima were found to be less effective. DTX-2 was discovered in contaminated Irish shellfish (46). 49).. 7. in Monterey Bay. and this is typically the method of choice. shellfish tissue can be efficiently extracted with aqueous methanol (1:1). The benthic Prorocentrum spp.
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.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.4. analysis of Prorocentrum cultures have uncovered an additional series of diol esters of OA and sulphated diesters (called DTX-4. In comparison.. This was probably not the first DSP episode. DSP toxins produced by benthic Prorocentrum spp. such as Prorocentrum lima. are also recognised as a threat to human health. and anecdotal evidence now suggests that several seafood poisoning episodes during the 1960s and 1970s in the Netherlands. 149 . Fig. Today over ten Dinophysis spp. Dinophysis caudata. and Dinophysis norvegica among others (47.FISH AND SHELLFISH TOXINS lipophilic compounds that are toxic in the mouse bioassay. 7. While these esters are not toxic in vitro. In this case Dinophysis was suspected as the culprit organism. Dinophysis acuta. but do not have the same mechanism of action as DSP toxins nor induce the same symptoms. are recognised as DSP toxin producers. DTX-5.5. The parent acids are the only DSP congeners to be found in shellfish tissue. 49). they can be converted to the parent toxins by nonspecific esterases or lipases either in the producing organism or in the host shellfish. and include Dinophysis acuminata. Norway and Japan were likely due to DSP toxins produced by a Dinophysis spp. Prorocentrum maculosum. 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. Dinophysis fortii. and DTX-6) all of which are rapidly converted to OA when the plankton cells are ruptured (48). which include the pectenotoxins and the yessotoxins. Structure of okadaic acid and the two related DSP toxins 7. and Prorocentrum belizeanum.
7. and while this provides critical seafood safety information. but the method is complex and matrix effects as well as interferences with other lipophilic compounds such as the pectenotoxins compromise the method. 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). At the first Meeting of the EU National Reference Laboratories on Marine Biotoxins and Analytical Methods and Toxicity Criteria in 1996. Preparation of a fluorescent 9-anthryldiazomethane (ADAM) derivative followed by HPLC offers reasonable sensitivity. The onset of symptoms can occur within 30 minutes of consuming contaminated shellfish and generally last about 2 . various European countries. nausea. and DTX-2. In recent years.2 Toxicology The DSP toxins are potent inhibitors of the serine/threonine phosphatases PP1 and PP2A. and a detection limit of 100 µg/kg has been reported (12). 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. 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. A regulatory level of 80 . 7. All the DSP toxins have around a thousandfold greater affinity for PP2A than PP1. even with the minimal of clean up steps (48). it does not provide information on the DSP toxin profile.5. Human symptoms include diarrhoea. Immunoassays have been developed for OA detection but an inherent problem of these methods has been the low cross reactivity with DTX-1. DSP episodes have been reported worldwide and are included in many monitoring programs (47). 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.FISH AND SEAFOOD Perhaps as a consequence of these multiple sources of DSP toxins. This regulatory level includes the possibility of LSPs in the DSP fraction being tested.3 Detection methods Once again the mouse or rat bioassay is relied upon to detect DSP toxins in contaminated seafood. The 7-O acyl derivatives (DTX-3) of OA that can form in shellfish also show poor cross reactivity and this further exacerbates the problem. vomiting.160 µg OA equivalents/kg of whole shellfish tissue has been implemented in some countries. some careful clean-up steps are required to avoid such matrix interferences and to simplify data interpretation (48). Due to the lipophilic nature of the toxins.3 days.5. Since the late 1970s. DSP episodes have been reported from Japan. and abdominal pain. New Zealand and Chile. 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). which can lead to an underestimation of the total DSP content in a sample. DSP-Check is a commercially 150 .
7. 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. fluorometric protein phosphatase inhibition assays have been found to perform better than colorimetric assays. Indeed. As a result these compounds are now being referred to as lipophilic shellfish toxins (LSTs) (49). they are toxic in the mouse bioassay and can result in false positives and hence are included here for completeness. 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.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. 58). However. with a claimed detection limit of 20 µg/kg.5. 7. with good agreement with the mouse bioassay and LC–based methods (54. 55). In yet another approach. and the latter group are usually found to be responsible for any reported shellfish toxicity (59). for many years these compounds were included in the DSP toxin complex.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). and another rapid antibodybased test kit for DSP toxins has also been reported (53). Structure of the principal pectenotoxins and one of the seco-acid derivatives 151 .5 and 7. over 20 PTXs are known (56) and more than 30 YTXs have been reported (57. Some representative examples of both groups of toxins are shown in Figures 7. Within this group. Fig.
FISH AND SEAFOOD Fig. YTX and homo-YTX 7. many parts of Europe and the Mediterranean. New Zealand and South America (49. 57. 7. 58).1 Origins and distribution It is now well recognised that PTXs are co-produced with DSP toxins by various strains of the dinoflagellate Dinophysis and hence the distribution of both families of toxins in shellfish is similar (49). which may co-occur in blooms with other organisms. but nevertheless the toxins are extremely widespread and have been found in various shellfish in Norway and Scandinavia. Structure of the two parent yessotoxins.6. Japan.2 Toxicology PTXs show varied toxicity in the mouse bioassay (59). eastern Canada. the toxin content and profile may vary among different strains. While 152 .6. As is typical for many algal species. 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). 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. 7. The sources of YTXs have now been identified as the dinoflagellates Protoceratium reticulatum and Lingulodinium polyedrum.6.
While LC/UV detection is possible for PTXs. Some of the PTXs are potent cytotoxins in vitro (60). The crude extract may be used directly in the mouse bioassay.6. 64). which is further compounded by differences in the experimental design.FISH AND SHELLFISH TOXINS demonstrating considerable toxicity following IP injection. 58). the PTXs show considerably less toxicity when administered orally. chemical analysis is best carried out by LC/MS and several approaches have been described (57. Although there is no evidence that yessotoxins are responsible for human illness. 7. but less so following oral dosing (61). There are two skeletal types known as PbTx-1 (or brevetoxin A) and PbTx-2 (brevetoxin B) (Figure 7. it may be compromised by other biological materials in the matrix. which aids detection by LC/UV. The YTXs are retained in the aqueous methanol fraction while the PTX and DSP toxins are concentrated in a dichloromethane fraction. The structures and chemical data including the molecular weight of each member have been collated (56). but a fluorescent derivative has been prepared for others. 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. For many years it was believed that 153 . though its cell effects may be linked to modulation of calcium channels (62). a regulatory level of 1 mg/kg shellfish tissue has been set. Acetone extracts of shellfish tissue are dried and re-suspended in 60% aqueous methanol before partitioning against dichloromethane. Many YTX toxins contain a conjugated diene system. Because of the number and complexity of the YTX group. The YTX group has not been reported to cause human illness. but various congeners are acutely toxic in the mouse bioassay following IP injection. Their toxicology and mechanism of action remain uncertain. 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 PTXs are susceptible to bioconversion in shellfish tissue and consequently it is often desirable to determine the profile of toxins present. There are no clear cases of human illness resulting from ingestion of PTXs (59). However. and this is believed to be due to hydrolysis of PTX-2 to the seco acid derivative. 7.7 Neurotoxin Shellfish Poisoning (NSP) Toxins NSP episodes are caused by a family of fused polyether compounds known as the brevetoxins (63.3 Detection methods Typically these toxins are co-extracted with DSP toxins following acetone or aqueous methanol extraction of the shellfish tissue.7). Estimates of acute toxicity of the parent compound following IP injection vary considerably. some of which are found as glycosides and sulfates.
the same toxins have been reported to be produced by three genera of Raphidophytes. which are found in Japanese waters (66). 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”. 7. namely Chatonella marina.7. They have also been associated with fish kills along the eastern mid-Atlantic coast of the US. Many of these species are associated with massive fish kills. Fig.7. In the Gulf of Mexico. Structure of the two parent brevetoxins. was chemically characterised as PbTx-2 (67). whelks. oysters. and one of the brevetoxin metabolites BTX-B2 7. Interestingly. and recently the major toxin associated with an ichthyotoxic unialgal bloom of Chatonella cf. brevetoxins are found in mussels.FISH AND SEAFOOD NSP toxins only occurred in the Gulf of Mexico and along the western coast of Florida. verruculosa off the mid Atlantic coast of the U. These are biotransformation products of principally PbTx-2 by addition of certain amino acids to the conjugated aldehyde function (66). 66). and off the coast of New Zealand (65). but in 1992/1993 they were identified as the culprits in a human poisoning event in New Zealand (65. and cockles (65.S. 66). PbTx-1 and PbTx-2. Fibrocapsa japonica.1 Origins and distribution The brevetoxins are produced by the dinoflagellate Karenia brevis (formerly known as Gymondinium breve and Pytochodiscus brevis) a dinoflagellate found in the Gulf of Mexico. and Heretosigma akashiwo. 154 .
malaise. The mean time to onset of symptoms is around 3 hours. 7. and in severe cases seizure. and mammalian body fluids without pretreatment or dilution (72). Exposure to brevetoxins can also occur through inhalation of brevetoxin aerosols resulting in irritation of the upper respiratory tract (65. which can result in a cascade of associated events in a cell (66). 66. 7. Ciguatera toxins (CTXs) are extremely potent and occur in finfish rather than shellfish. The mouse bioassay is used as the primary detection method. The brevetoxin metabolites are reported to be less toxic than the parent compound (70). this family of toxins has proven to be a challenge both to identify and monitor. with a mean duration of symptoms of around 1 day. but this method is now used routinely in New Zealand and the US to monitor for the presence of brevetoxins (68). the competitive ELISA method correlated most closely with the mouse bioassay results. shellfish homogenates. In a comprehensive multi laboratory evaluation of five different detection methods as possible replacements for the mouse bioassay. with hot spots in the coral reef areas of the Caribbean and French Polynesia.7. Outbreaks of ciguatera poisoning are confined to tropical and sub-tropical regions of the ocean. 69). In this application.7.3 Detection methods Three basic approaches for the detection of NSP toxins have been recently reviewed (71). pre-treatment or clean-up of the shellfish sample is essential in order to obtain reproducible results. and it has also been used in studies to quantify occupational and recreational exposure to aerosolised brevetoxin exposure (73). and an action level has been set at 800 µg/kg (or 80 µg PbTx-2 equivalents/100 g or 20 MU/100 g or 4 µg/mouse) of shellfish tissue to trigger regulatory action. The human health implications following exposure to NSPs have been reviewed (69).8 Ciguatoxins Known for centuries and originally referred to as ciguatera by Spanish explorers in the Indo-Pacific ocean and Caribbean.2 Toxicology Brevetoxins bind to site 5 of voltage-gated sodium channels. A second generation competitive ELISA assay has been developed that has successfully quantified brevetoxins in a variety of biological and environmental matrices including seawater.FISH AND SHELLFISH TOXINS although recently it has been shown that herbivorous fish can store brevetoxins in their flesh (68). 7. Ingestion of brevetoxins results in a variety of symptoms including parathesia. While the mouse bioassay continues to be used. considerable progress has been made in the development of LC-MS methods to routinely monitor for brevetoxins in shellfish. vertigo. nausea. beginning with herbivores that feed on benthic dinoflagellates associated with reefs and which are 155 . and diarrhoea.
FISH AND SEAFOOD transferred through the food chain to the larger carnivores. Fig. and only a few structures have been identified. Indeed even the chemistry of CTXs from the Pacific differs subtly from those found in the Caribbean (Figure 7.8. Such microalgae contribute to the primary production of tropical and sub-tropical reefs and live in epiphytic association with bushy red. 7. A detailed comprehensive review of ciguatoxins has recently been published (74). complex. most likely a consequence of biotransformation of the parent toxins as they pass up the food chain. but more are suspected to exist. These include species belonging to the genera Ostreopsis and Coolia. a group of closely related polyethers that are considered the source of ciguatoxins. The diversity of benthic phytoplankton that exist on reefs further complicates the picture.8. although the dinoflagellate Gambierdiscus toxicus is considered to be the major source of these toxins. and other genera of neighbouring reef dinoflagellates can further contribute to an extremely complex profile of toxins in finfish. Structure of the Pacific variant of ciguatera toxin (P-CTX-1) and the Caribbean variant (C-CTX-1) 7.1 Origins and distribution It is recognised that several species of benthic dinoflagellates can produce CTXs and the gambiertoxins. Culprit species producing CTXs may simultaneously produce unrelated toxins such as maitotoxin. lipid soluble polyethers. Distribution through the food chain begins with 156 . CTXs are large. green and brown seaweeds and also occur in sediments and coral debris.8).
the neurological symptoms can persist for days. Local island populations attempt to avoid poisoning by catching only smaller fish. and inducing membrane depolarisation resulting in prolonged symptoms indicative of damage to nerve tissue. weeks or even months (74 .3 Detection methods As a consequence of their potency. Other symptoms include a metallic taste. Another symptom that can develop over time is extreme fatigue. which can return following the consumption of alcohol and certain foods such as oily fish (74. but fish toxicity can vary widely depending on where the fish are caught and how long they have been feeding in that area. which in turn are consumed by carnivores such as snappers. the absence of a strong chromophore means that simple detection methods based on UV or fluorescence-based detection methods are ineffective. arthralgia. and moray eels. A good knowledge and careful observation of the symptoms in the bioassay (Minimal 157 . all of which can be linked to the effect of CTX on excitable membranes.8. 7. barracudas. 75). The onset of symptoms can occur rapidly within 24 hours. and it is estimated that ingestion of as little as 0. Even when suspect samples are available. The pharmacological effects following CTX exposure are numerous and include severe gastrointestinal distress and neurological symptoms. 78). These include paresthesias (numbness and tingling of the extremities). Patients have reported symptoms lasting weeks and even months. which can be confused with chronic fatigue syndrome.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. and in fact CTX poisoning is often only identified after consumption of contaminated fish. though the mouse bioassay continues to be relied upon as the most common method of detecting CTXs.FISH AND SHELLFISH TOXINS grazing by herbivorous fish including the sturgeon fish and parrotfish. 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. 7. 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. sharks. although not all CTXs are equipotent. groupers.76). where cold objects feel hot and vice versa. myalgia. Differences in potency among the CTXs correlate directly with their binding affinity for the sodium channel. and the sensation of loose teeth (77). only trace amounts of CTXs can cause illness following consumption of contaminated fish. 75. and temperature sensation reversal. Like the brevetoxins. Globally. there are neither standards nor an official testing program.8. pruritus. they bind to voltage-dependant sodium channels in the cell membrane causing them to open. and although the gastronomical problems may dissipate within a day or so.
Acetone extracts of fish tissue are subjected to solvent partitioning. which features characteristic fragment ions corresponding to sequential losses of three molecules of water. and another dealing with the chemistry and detection methods has been published (85).g. Usually the major AZA derivatives found in shellfish are AZA1 (50 . although specificity and sensitivity remain major hurdles (74) and the presence of DSP toxins affects the test (80). The original structure proposed revealed an entirely new class of shellfish toxin.80%). Immunoassays may be an alternative detection method (79). and only a few structures (AZA 1 . 158 .FISH AND SEAFOOD Lethal Dose (MLD) 0.85).25 µg/kg) can provide useful clues as to the toxins present but more specific and informative assays are required. and following synthesis of the molecule some time later. and the corrected structure was published in 2004 (86) (Figure 7. Since the original discovery and identification of the toxin. almost 20 additional derivatives have been reported (83 . specific for CTX activity.30%) and AZA3 (5 . following consumption of contaminated mussels Mytelius edulis (82). which are based on an LC-MS/MS approach.20%) present in lesser amounts (Figure 7. and it is likely that most of these compounds are biotransformation products formed in the shellfish tissue. This is then subjected to LCMS/MS analysis. followed by clean-up on silica and aminopropyl cartridges. In this way Caribbean and Pacific CTX-1 compounds can be distinguished (74).9 Azaspiracids Azaspiracid (AZA1) was originally identified in 1995 as the causative agent for a poisoning episode in Ireland. with AZA2 (10 . Recent reviews of this new group of toxins have appeared (83. 7. high-throughput screen (74). A sodium channel-specific assay using mouse neuroblastoma cells has been described although this approach has still to be developed in a routine.9). Chemical detection methods are severely compromised by the lack of standards.5) have been fully characterised by Nuclear Magnetic Resonance (NMR) and MS analysis. at C-3 and C-23). and the separation of these polyethers from the lipid-soluble fraction of fish provides an additional challenge. 84). only a few laboratories are able to undertake such analyses. minor revisions of a few stereocenters were made.9). Usually the additional AZA derivatives found in shellfish tissue are hydroxylated derivatives (e. but this only has application in a clinical setting (81). Consequently. has been developed. another in vitro sodium channel-based assay. Recently. The majority of these have been proposed based on LC/MS analytical data.
clams (Tapes philipinarum. There has been considerable debate over the source of azaspiracids. 7. and most recently in a crustacean (Cancer pagurus). Structure of the three principal azaspiracids found in shellfish 7. but current data indicates that unlike the DSP compounds.FISH AND SHELLFISH TOXINS Fig.1 Origins and distribution The azaspiracids were first isolated from contaminated shellfish. 84.). scallops (Pecten maximus). AZA occurrence has been reported by several European countries including France. vomiting. cockles (Cerastoderma edule). AZA toxins do 159 . cassipes produces AZAs in culture. While this result is not necessarily in debate. The bivalves affected include mussels (Mytilus edulis. which can persist for 2 . oysters (Crassostrea gigas.9. Ostrea edulis). In fact the routes of trophic transfer of AZAs through the food chain ending in contaminated shellfish remain unknown. 7. These symptoms are not unlike those experienced following the consumption of DSP toxins. Scandinavia. and some time after the original episode in Ireland. cassipes (83). and to date this has been the only source from which these toxins have been found at toxic levels. Portugal. 88. which are ingested by P. However. these molluscs were never considered to be the biogenetic source. it has been impossible to establish that P. diarrhoea and stomach cramps. Following the original episode. it was reported that the heterotrophic dinoflagellate Protoperidinium cassipes was the planktonic source of the toxins (87). Donax spp. and off the coast of Morocco (83. and to date no long-term effects have been recorded (83. 90). 89).3 days.9. Ensis siliqua. and variable field results have led to suggestions that the actual producing organisms are Dinophysis spp.9. Mytilus galloprovincialis).2 Toxicology Consumption of AZA contaminated shellfish can result in acute symptoms that include nausea. as in other cases of shellfish toxins.
Key fragment ions are useful in identifying the various AZA compounds that can occur in shellfish tissue (83. and erosion and bleeding in the stomach. 7. and necrosis of epithelial cells on the microvilli (91). However. which are particularly amenable to detection by positive ion electrospray mass spectrometry. parenchymal cell pyknosis in the pancreas. Due to their relatively recent discovery and limited knowledge of their toxicological properties and impact on human 160 . AZAs also display cytotoxicity towards various mammalian cell lines (83. the spirolides. This common cyclic imine feature is believed to be a common pharmacophore of this group of toxins which comprises the gymnodimines. The mouse bioassay has been used to monitor shellfish extracts. it is believed that the hydroxylated derivatives AZA4 (470 mg/kg) and AZA5 (<1000 mg/kg) are considerably less toxic. and ASP. Importantly these pathological outcomes were reported as being different from those caused by DSP. 91. Injection of a lethal dose of AZA1 in mice (>150 µg/kg) caused vacuole formation and fatty acid accumulation in hepatocytes. 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. The limited amounts of AZA toxins available continue to hinder a complete understanding of their mechanism of action. Oral administration of AZAs at substantially higher doses than those used in IP injection studies did not result in any mouse mortalities after 24 hours. and closely related ptierotoxins (Figures 7. PSP. though the possibility exists that they may inhibit other phosphatases.9.FISH AND SEAFOOD not inhibit the phosphatases PP1 and PP2.3 Detection methods The regulatory level for AZAs in shellfish has been set by the EU at 160 µg AZA equivalents/kg shellfish tissue. abnormalities in the thymus and spleen.10 Cyclic imines These toxins are represented by several distinct structural types. 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).12). 7. Based on IP minimum lethal dose data. 85). but all contain a spirocyclic or cyclic imine function within their structure. the pinnatoxins. 92). Once again. it is suggested that the order of toxicity is AZA2 (110 µg/kg) >AZA3 (140 µg/kg) >AZA (150 µg/kg). 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. Using the same approach. although autopsies conducted after 4 and 8 hours revealed various gastrointestinal (GI) abnormalities including accumulation of fluid in the ileum.10 – 7. Currently LC/MS and LC-MS/MS analysis is the preferred chemical analytical detection method for AZAs. but unlike DSP monitoring methods which use only the hepatopancreas of the shellfish for extraction.
The gymnodimines are the smallest members of the cyclic imine group with a molecular weight range of 500 . 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. 95). 7. selliformis (97).10). scallops.10. Subsequent taxonomic studies revised this again to a new dinoflagellate K. 95). At the time of the initial observation of gymnodimine. Fig.10. Reviews of the current status of the cyclic imine group were published by Molgo and Cembella (94. Some time later. gymnodimine B and C were isolated from the dinoflagellate Karenia selliformis (formerly Gymnodinium selliformis). but they are included here for completeness.. cockles. led to the identification of a new marine toxin with an unprecedented structure named gymnodimine (96) (Figure 7. it was proposed that a dinoflagellate identified as Gymnodinium spp. they are sometimes referred to as emerging toxins of uncertain human health concern. 7.FISH AND SHELLFISH TOXINS health. In only two cases (gymnodimines and spirolides) has a planktonic source been identified. two additional congeners. a species closely related to Gymnodinium mikimoitoi was the planktonic source of the toxin.1. 161 . The structure is novel in that it contains a spirocylic imine group and a single tetrahydrofuran ring embedded within a macrocyclic structure.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 (94.1 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. surf clam and New Zealand abalone. Structure of the cyclic imine gymnodimine 7. Pacific oyster.
Whether isolated from dinoflagellate cells directly or from shellfish tissue. it has been stated that regulation against gymnodimine is not required (98).10. and this is the detection method of choice. finishing with 100% methanol. 7. 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. abdominal breathing. and so spectroscopic detection by UV methods is not favoured.11). 95) (Figure 7. Most cyclic imines lack an extended chromophore. Since their original discovery which resulted in the characterisation of spirolides A-F (99. Fractions are then dried and re-suspended in methanol or acetonitrile in preparation for LC/MS analysis (94.3 Detection methods The characteristic “fast-acting. and short-acting cholinesterase inhibitors such as neostigmine or physostigmine protected mice against the toxic effects of gymnodimine.1. extracts exhibited the fast acting symptoms that became associated with cyclic imines following IP injection. 50%) of 96 µg/kg. cyclic imines are readily observed by electrospray mass spectrometry in the positive ion mode.2 Toxicology A review of the toxicology of all cyclic imines has been published (98).100). edulis) and scallops (Placopectin magellanicus) from eastern Canada that displayed abnormal symptoms in the mouse bioassay. In particular. The original observation of toxicity in New Zealand followed from IP injection of lipophilic extracts in the mouse bioassay. Reduction of the imine group eradicated the toxicity. However.FISH AND SEAFOOD 7.to10-fold reduction of toxicity (LD50 600 . Based on this diminished oral toxicity. This led to the suggestion that gymnodimine acts by blocking nicotinic acetylcholine receptors at the neuromuscular junction. 95).900 µg/kg). Oral administration of the toxin to mice by gavage resulted in about an 8.10. almost a dozen derivatives in total have been identified to date (94. A later study using purified toxin established an LD50 dose (Lethal Dose. The structure of the spirolides bears a strong resemblance to both gymnodimine and 162 . and lack of responsiveness to stimuli.10. The time lapse between ingestion and death is between 5 and 15 minutes.2 Spirolides The spirolides comprise the largest group of cyclic imine compounds.1. 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. 7. all or nothing” mouse bioassay response to gymnodimines and cyclic imines in general continues to be a useful monitoring approach. There is a rapid onset of symptoms including respiratory distress. together with a lack of evidence of human intoxication.
by gavage (87 .1 Origins and distribution At the time of the original discovery of the spirolides in shellfish from the coastal waters of eastern Nova Scotia.2. in fjords in Denmark and Norway. All data reported here are for desmethyl spirolide C. Fig.11. This proved to be the case and eventually the dinoflagellate Alexandrium ostenfeldii was identified as the culprit organism (101). Structural differences among the various spirolides result from variations in the number of methyl groups and the sequence of spiroketal rings 6-5-5 in one group and 6-6-5 in the other.2 Toxicology The spirolides are the most studied group of the cyclic imines. Since then the spirolides have been found to be globally distributed and are found in shellfish or plankton fractions along eastern Canada.10. these are the toxins that are also most resistant to acid hydrolysis of the imine function 163 .11. the Bay of Biscay and most recently from Mediterranean shellfish and plankton in the Adriatic (102).FISH AND SHELLFISH TOXINS the pinnatoxins (vide infra). the gulf of Maine. those spirolides possessing the vicinal dimethyl grouping (see Figure 7.10. southern Ireland.2. 7. the western coast of Scotland. their localisation in the digestive gland of mussels and scallops led to the suspicion that plankton were the real source of the toxins. Interestingly. Structure of four spirolide toxins A-D 7.707 µg/kg. 7. fasted mice). or by oral ingestion (381 .166 µg/kg. R1 = CH3) of the cyclic imine ring are the most toxic of the group (98). fasted mice). While oral toxicity is considerably less than by IP injection.8 µg/kg). The LD50 values vary considerably depending upon whether the compound is administered by IP injection (5 . 95). They have also been reported in the southern hemisphere in Chilean mussels and in New Zealand shellfish and plankton (94.
The compounds are zwitterionic and therefore somewhat more polar than the spirolides. A number of methods following this procedure have been described (94. While the mechanism of action remains to be clearly defined.3 Detection methods As discussed for gymnodimine.FISH AND SEAFOOD (100). which includes two diastereomers. more useful information on the amount and distribution of toxins present in a shellfish extract is obtained by LC-MS analysis usually following a simple clean-up step. 7. Fig. 7.3 Pinnatoxins and pteriatoxins Pinnatoxins A . Structure of the pinnatoxin group and the closely related pteriatoxins 164 . 95. 7.2.B contain a spiro imine moiety similar to that found in the spirolides. consistent with modulation of acetyl cholinergic receptors (103). and in fact these compounds share a number of other structural features.12). Three members of this group have been reported. To date. The pteriatoxins were isolated from a different mollusc but bear a strong structural resemblance to the pinnatoxins (Figure 7. the mouse bioassay can be used to detect these fast acting toxins.12.10. 102). 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. However.10. Once again this relies on the sensitivity of these cyclic imines to detection by positive ion electrospray mass spectrometry.
there is no compelling evidence to suggest that they have resulted in human toxicity (98). This prompted a chemical investigation of Pinna muricata that ultimately led to the identification of the pinnatoxins in 1995 (104). and in some cases it is known that one toxin can potentiate or augment the toxicity of the other. have long been suspected of causing human illness after ingestion.most likely a dinoflagellate. Coupled with this. and more advanced means of detection. The LD99 dose for the pteriatoxins is reported to be in the range (8 . and later the closely related pteriatoxins were isolated from the digestive glands of another Okinawan bivalve Pteria penguin (105). The LD99 (Lethal Dose.3. new toxin groups such as the azaspiracids and cyclic imines have also been reported. once again reflecting improvements in purification and spectroscopic methods permitting structural identification of new and complex structures.10. 7. However. like the gymnodimines and spirolides.10. Only the acute toxicity in mice following IP injection has been reported. With increasing knowledge of the chemistry of toxins.3.10. 99%) values for pinnatoxins A-C are in the range (22 . While this is recognised as a real possibility for a wide range of toxins.3 Detection methods No formal detection methods for these compounds have been reported. while pinnatoxin D was essentially inactive. but this remains to be conclusively established (106). to date action or 165 .3. Despite this progress. which in many cases are now found to be more widespread than previously believed.1 Origins and distribution Shellfish of the genus Pinna. but the mouse bioassay data and LC-MS can be used to monitor suspect seafood for these compounds. 7.100 µg/kg). This is generally a result of increased monitoring and continually improving detection technologies and methods. which occupy tropical waters around Okinawa and the south east coast of China. 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.2 Toxicology There is little information describing the toxicology of these compounds (98).180 µg/kg). There is strong circumstantial evidence to suspect that these toxins. has come the realisation that a “toxin profile” can be a complex mixture of many different classes of toxins.FISH AND SHELLFISH TOXINS 7. arise from an algal source .11 Conclusion There has been considerable progress in the detection of known marine toxins. 7.
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Bienfang P. Benson J. Henry M.R. Manzo L.. 171 .C. 187-94. Botana L. in Seafood and Freshwater Toxins. Fleming L.G. Detection of brevetoxins in the twenty-first century. Ciguatera fish poisoning in industrial ship crew members: a retrospective study in a seaport general practice in Trinidad and Tobago. CRC Press. 551-8. Competitive ELISA to detect brevetoxins from Karenia brevis (formerly Gymnodinium breve) in seawater. Schrank K. Naar J..A. Ed.J. Wanner A. 50. Tomas C.E.. 99-115. 71. 53. Freidman M. Food and Agriculture Organisation.. Literature review of Florida red tide: implications for human health.S. Marine Drugs. Bourdelais A.A.M. 707-23. Jacocks H.P. 131.W. Gannon D...C. Wolny J. 3. 2008. Tomas C.. 2007.. Ciguatera fish poisoning: treatment prevention and management. Granholm A... Cheng Y. Dechraoui M-Y..H.M. 2000. 117-71..... Baden D. Prockop L.A.. Kubanek J. 220-6.. Ed.. 825-34. Zaias J... Whitney P. 73. 76.. like ciguatoxins... Kirkpatrick B. Toxicon.. 6. Pierce R. Boca Raton. Landsberg J. Ed.. Botana L. Buonocore M.. and detection. Bean J.S. Lancaster J. Locatelli C. Butera R. Chen A. Gandini C. 110. Pierce R..M. Flewelling L. 23. Wang Z.E..E. 179-85. The molecular and integrative basis to brevetoxin toxicity. 110. 77. 50. Abraham W. Harmful Algae.S. Abbott J. 2004. Aerosolised red tide toxins (Brevetoxins) and asthma. 456-79. Food and Agriculture Organisation of the United Nations. Bossart G.. Naar J. Lenzi A. New fish-killing alga in coastal Delaware produces neurotoxins.. in Seafood and Freshwater Toxins.. Baden D. Poli M.. CRC Press. Poon-King T. Fleming L.P. shellfish and mammalian body fluid... 66. Neurologic Shellfish Poisoning (NSP).. are potent ichthyotoxic neurotoxins that accumulate in fish. Baden D. 519-50. 2008. Rome.M..FISH AND SHELLFISH TOXINS 65. West Indian Medical Journal. Environmental Health Perspectives. Kubanek J. Bourdelais A.J. Cheng Y.N. in Seafood and Freshwater Toxins.P. 74. 70.. CRC Press. 68.... Mild ciguatera poisoning: case reports with neurophysiological evaluations. Steidinger K.A. 75. Watkins S. Backer L. Toxicon.. Kirkpatrick B.. Weisman R..S. Fleming L.. Chest. Marine Biotoxins. Ed. Zaias J. 2008..G.. Fernandez M.D. toxicology. Abraham W. 465-70. Boca Raton. Johnson D. Botana L.. Dickey R. Naar J.. Ayyar R. Baden D.E.B.. Reich A.. 2004. in FAO Food and Nutrition Paper 80. Dickey R. Poon-King C. Flewelling L. Ramsdell J. Naar J. 2002. Clark R.... Reich A.. Baden D.. Backer L... Backer L. Pierce R.. 69. Granade R.. 2007. 67. 2002. 1598-603. Boca Raton.G. Squicciarini D. Bottein M-Y..D. Ramsdell J. Ciguatera toxins: Chemistry. 479-516..G.M. 2004.E.. 2007.G. Muscle Nerve. Intrinsic potency for synthetically prepared brevetoxin cysteine metabolites BTX-B2 and desoxy-BTX-B2... Food and Agriculture Organisation. Environmental Health Perspectives.. 72... Brevetoxins. 2008.
FISH AND SEAFOOD 78. 79. Galziou P., Marti P.M.V. Study of factors that influence the clinical response to ciguatera fish poisoning. Toxicon, 1993, 31, 1151-4. Hokama Y., Ebesu J.S.M., Nishimura K., Oishi S., Mizuo B., Stiles M., Sakamoto B., Takenaka W., Nagai H. Human intoxication from Hawaiian reef fishes associated with diverse marine toxins. Journal of Natural Toxins, 1996, 5, 235-47. Park D.L. Detection of ciguatera and diarrheic shellfish toxins in finfish and shellfish with Ciguatect kit. Journal of AOAC International, 1995, 78, 533-7. Bottein-Dechraoui M.Y., Wang Z., Turquet J., Chinain M., Darius T., Cruchet P., Radwan F.F., Dickey R.W., Ramsdell J.S. Biomonitoring of ciguatera exposure in mice using blood collection cards. Toxicon, 2005, 46, 243-51. Satake M., Ofuji K., Naoki H., James K.J., Furey A., McMahon T., Silke J., Yasumoto T. Azaspiracids, a new marine toxin having unique spiro ring assemblies, isolated from Irish mussels Mytilus edulis. Journal of the American Chemical Society, 1998, 120, 9967-8. Twiner M.J., Rehmann N., Hess P., Doucette G.J. Azaspiracid Shellfish Poisoning: A review of the chemistry, ecology and toxicology with an emphasis on human health impacts. Marine Drugs, 2008, 6, 39-72. Food and Agriculture Organisation of the United Nations. Azaspiracid Shellfish Poisoning (AZP), in FAO Food and Nutrition Paper 80. Marine Biotoxins. Ed. Food and Agriculture Organisation of the United Nations. Rome, Food and Agriculture Organisation. 2004, 173-84. James K.J., O’Driscoll D., Fernandez J.G., Furey A. Azaspiracids: chemistry, bioconversion and determination, in Seafood and Freshwater Toxins. Ed. Botana L.M. Boca Raton, CRC Press. 2008, 763-73. Nicolao K.C., Koftis T.V., Vyskocil S., Petrovic G., Ling T., Yamada T.M.A., Tang W., Frederick M.O. Structural revision and total synthesis of azaspiracid-1, Part 2: Definition of the ABCD domain and total synthesis. Angewandte Chemie International Edition, 2004, 43, 4318-24. James K.J., Moroney C., Roden C., Satake M., Yasumoto T., Lehane M., Furey A. Ubiquitous ‘benign’ alga emerges as the cause of shellfish contamination responsible for the human toxic syndrome azaspiracid poisoning. Toxicon, 2003, 41, 145-51. Magdalena A.B., Lehane M., Krys S., Fernandez M.L., Furey A., James K.J. The first identification of azaspiracids in shellfish from France and Spain. Toxicon, 2003, 42, 105-8. Vale P., Bire R., Hess P. Confirmation by LC-MS/MS of azaspiracids in shellfish from the Portuguese north-western coast. Toxicon, 2008, 51, 1449-56. Ryan G., Cunningham K., Ryan M.P. Pharmacology and Epidemiological impact of azaspiracids, in Seafood and Freshwater Toxins. Ed. Botana L.M. Boca Raton, CRC Press. 2008, 755-61. Ito E. Toxicology of azaspiracid-1: Acute and chronic poisoning, tumorigenicity, and chemical structural relationship to toxicity in a mouse model, in Seafood and Freshwater Toxins. Ed. Botana L.M. Boca Raton, CRC Press. 2008, 775-84.
FISH AND SHELLFISH TOXINS 92. Twiner M.J., Hess P., Bottein-Dechraoui M-Y., McMahon T., Samons M.S., Satake M., Yasumoto T., Ramsdell J.S., Doucette G.J. Cytotoxic and cytoskeletal effects of azaspiracids-1 on mammalian cell lines. Toxicon, 2005, 45, 891-900. Moroney C., Lehane M., Brana-Magdalena A., Furey A., James K.J. Comparison of solid phase extraction methods for the determination of azaspiracids in shellfish using electrospray liquid chromatography-mass spectrometry. Journal of Chromatography, 2002, 963, 353-61. Molgo J., Girard E., Benoit E. Cyclic imines: An insight into this emerging group of bioactive marine metabolites. Phycotoxins, 2007, 319-35. Cembella A., Krock B. Cyclic imine toxins: Chemistry, biogeography, biosynthesis and pharmacology, in Seafood and Freshwater Toxins. Ed. Botana L.M. Boca Raton, CRC Press. 2008, 561-80. Seki T., Satake M., MacKenzie L., Kaspar H., Yasumoto T. Gymnodimine, a new marine toxin of unprecedented structure isolated from New Zealand oysters and the dinoflagellate Gymnodinium sp. Tetrahedron Letters, 1995, 36, 295-9. Haywood A.J., Steidinger K.A., Truby E.W., Bergquist P.I., Adamson J., Mackenzie L. Comparative morphology and molecular phylogenetic analysis of three new species of the genus Karenia (Dinophyceae) from New Zealand. Journal of Phycology, 2008, 40, 165-79. Munday R. Toxicology of cyclic imines: Gymnodimine, spirolides, pinnatoxins, pteriatoxins, prorocentrolides, spiro-centrimine, and symbioimines, in Seafood and Freshwater Toxins. Ed. Botana L.M. Boca Raton, CRC Press. 2008, 581-94. Hu T., Curtis J.M., Oshima Y., Quilliam M.A., Walter J.A., Watson-Wright W., Wright J.L.C. Spirolides B and D, two novel macrocycles isolated from the digestive glands of shellfish. Journal of the Chemical Society, Chemical Communications, 1995, 2159-61.
100. Hu T., Burton I.W., Cembella A.D., Curtis J.M., Quilliam M.A., Walter J.A., Wright J.L.C. Characterisation of spirolides A, C, and 13-desmethyl C, new marine toxins isolated from toxic plankton and contaminated shellfish. Journal of Natural Products, 2001, 64, 308-12. 101. Cembella A.D., Lewis N.J., Quilliam M.A. The marine dinoflagellate Alexandrium ostenfeldii (Dinophyceae) as the causative organism of spirolide shellfish toxins. Phycologia, 2000, 39, 67-74. 102. Ciminiello P., Dell’Aversano C., Fattorusso E., Magno S., Tartaglione L., Cangini M., Pompei M., Guerrini F., Boni L., Pistocchi R. Toxin profile of Alexandrium ostenfeldii (Dinophyceae) from the northern Adriatic Sea as determined by liquid chromatography-mass spectrometry. Toxicon, 2006, 47, 597-604. 103. Gill S., Murphy M., Clausen J., Richard D., Quilliam M., MacKinnon S., LeBLanc P., Mueller R., Pulido O. Neural injury biomarkers of novel shellfish toxins, spirolides: a pilot study using immunochemical and transcriptional analysis. Neurotoxicology, 2003, 24, 593-604. 104. Uemura D., Chou T., Haino T., Nagatsu A., Fukuzawa S., Zheng S.Z., Chen H. Pinnatoxin A: a toxic amphoteric macrocycle from the Okinawan bivalve Pinna muricata. Journal of the American Chemical Society, 1995, 117, 1155-6. 173
FISH AND SEAFOOD 105. Takeda N., Uemura N., Suenaga K., Uemura D. Structural determination of pteriatoxins A, B, C, extremely potent toxins from the bivalve Pteria penguin. Tetrahedron Letters, 2001, 42, 3495-7. 106. Kuramoto M., Arimoto H., Uemura D. Bioactive alkaloids from the sea. Marine Drugs, 2004, 1, 39-54.
8. HACCP IN FISH AND SEAFOOD PRODUCT MANUFACTURE
Rhea Fernandes and Dr. Peter Wareing Leatherhead Food International Randalls Road Leatherhead Surrey KT22 7RY United Kingdom 8.1 Introduction
The Hazard Analysis Critical Control Point (HACCP) system is a structured, preventative approach to ensuring food safety. HACCP provides a means to identify and assess potential hazards in food production and establish preventive control procedures for those hazards. A critical control point (CCP) is identified for each significant hazard, where effective control measures can be defined, applied and monitored. The emphasis on prevention of hazards reduces reliance on traditional inspection and quality control procedures and end-product testing. A properly applied HACCP system is now internationally recognised as an effective means of ensuring food safety. The HACCP concept can be applied to new or existing products and processes, and throughout the food chain from primary production to consumption. It is compatible with existing standards for quality management systems such as the ISO 9000-2000 series, and HACCP procedures can be fully integrated into such systems. The new ISO 22000 food safety standard formally integrates HACCP within the structure of a quality management system. HACCP is fully integrated into the British Retail Consortium (BRC) Global Standard for Food Safety, and is one of the ‘fundamental’ requirements of that system. The application of HACCP at all stages of the food supply chain is actively encouraged, and increasingly required, worldwide. For example, the Codex Alimentarius advises that "the application of HACCP systems can aid inspection by regulatory authorities and promote international trade by increasing confidence in food safety". In many countries, there is a legal requirement for all food business operators to have some form of hazard analysis based on HACCP as a means of ensuring food safety. For example, within the European Union, Regulations 852/2004 and
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853/2004 require a fully operational, and maintained HACCP system, according to Codex, to be in place. 8.2 Definitions
Control (verb) - To take all necessary actions to ensure and maintain compliance with criteria established in the HACCP plan. Control (noun) - The state wherein correct procedures are followed and criteria are met. Control measure - An action and activity that can be used to prevent or eliminate a food safety hazard or reduce it to an acceptable level. Corrective action - An action to be taken when the results of monitoring at the CCP indicate a loss of control. Critical Control Point (CCP) - A step at which control can be applied and is essential to prevent or eliminate a food safety hazard, or reduce it to an acceptable level. Critical limit - A criterion which separates acceptability from unacceptability. Deviation - Failure to meet a critical limit. Flow diagram – A systematic representation of the sequence of steps or operations used in the production or manufacture of a particular food item. HACCP - A system which identifies, evaluates and controls hazards which are significant for food safety. HACCP Plan – A document prepared in accordance with the principles of HACCP to ensure control of hazards which are significant for safety in the segment of the food chain under consideration. Hazard - A biological, chemical or physical agent in, or condition of, food with the potential to cause an adverse health effect. Hazard analysis - The process of collecting and evaluating information on hazards and the conditions leading to their presence to decide which are significant for food safety and therefore should be addressed by the HACCP plan. Monitoring – The act of conducting a planned sequence of observations or measurements of control parameters to assess whether a CCP is under control.
procedures. 4. operation or stage in the food chain including raw materials. The team should also include 177 . 7. Establish procedures for verification to confirm that the HACCP system is working effectively.Obtaining evidence that the elements of the HACCP plan are effective. Establish the critical limit(s).A point. Determine the CCPs. 6.HACCP IN FISH AND SEAFOOD PRODUCT MANUFACTURE Step . Each stage is discussed below in detail. 8. Validation . procedure. 3. 5.3.3 Stages of a HACCP Study The HACCP system consists of the following seven basic principles: 1.1 Assemble HACCP team HACCP requires management commitment of resources to the process. quality assurance or control. tests and other evaluations. An effective HACCP plan is best carried out as a multidisciplinary team exercise to ensure that the appropriate product-specific expertise is available. Establish a system to monitor control of the CCP. food chemistry or engineering. 8. Figure 8. hygiene managers. 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.1 is a flow diagram illustrating this 12-stage logic sequence.The application of methods. in addition to monitoring to determine compliance with the HACCP plan. from primary production to final consumption. Establish documentation concerning all procedures and records appropriate to these principles and their application. Establish the corrective action to be taken when monitoring shows that a CCP is not under control. 2. 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. food microbiology. ingredient and packaging buyers. Conduct a hazard analysis. Verification .
If expert advice is not available on-site. 8. it may be obtained from external sources. biological.g. chemical and/or physical).1. 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. Logic sequence for application of HACCP 178 . Fig.FISH AND SEAFOOD personnel who are involved with the variability and limitations of the operations.
It should provide an accurate representation of all steps in the production process from raw materials to the end179 . 8. 8.4 Construct a flow diagram The flow diagram should be constructed by the HACCP team and should contain sufficient technical data for the study to progress. It is also important to identify the consumer target groups. including practices onboard fishing vessels.3. heat treatment. challenge testing and inoculation studies for evaluation of safety aspects are therefore essential for an effective HACCP team. etc. packaging. 8. aquaculture drug residues and potentially pathogenic Vibrio spp. may need to be considered specifically. storage and distribution conditions and instructions for use.1 Fish and seafood Certain products may be contaminated or carry pathogenic organisms as a part of the natural flora. or in aquaculture operations. etc. Vulnerable groups of the population.3 Identify intended use The intended use should be based on the expected uses of the product by the enduser or consumer (e. for example contamination by foodborne pathogens as a result of time/temperature abuse or unsatisfactory handling practice.3. it must be noted here that certain toxins (algal and shellfish) are not destroyed by heat treatments. such as enteric viruses in shellfish. If the processing does not include a kill step. physical and chemical structure (including water activity (aw).2 Describe the product It is important to have a complete understanding of the product. fermentation. In addition. These factors are likely to have a significant effect on some potential hazards. such as children or the elderly. naturally present on fish from warm waters (> 15 °C). which should be described in detail. there may be species-specific hazards related to certain fish and shellfish. marine toxins associated with particular fish and shellfish species.3.3. freezing. pH.1.3. 8.g. The description should include information such as the product name. Expertise on such inherent hazards. is a cooking process required?). processing conditions (e.).).3.HACCP IN FISH AND SEAFOOD PRODUCT MANUFACTURE 8.1 Fish and seafood The HACCP team should ideally have access to expertise on the appropriate fisheries. shelf life. the only CCP that can render the product safe from pathogenic organisms is adequate heat treatment during preparation.g. contaminated fish and shellfish should not be used. However. composition.
and identify any measures to control identified hazards The HACCP team should list all hazards that may reasonably be expected to occur at each step in the production process. ingredient specifications. and therefore a detailed description of the procedure applied must be available. 8. If HACCP studies are applied to proposed new process lines or products. The operation should be observed at all stages. adequate chilling of finfish immediately after capture is vital for the control of several potential hazards.1 Fish and seafood Examples of flow diagrams for specific fish products may be found in the appropriate product chapters. The analysis is likely to include consideration of: ● the likely occurrence of hazards and the severity of their adverse health effects. survival or multiplication of pathogenic microorganisms. conduct a hazard analysis. cleaning and hygiene procedures and storage conditions. it should also include details of CCP steps.3. Many fish-processing operations have relatively few steps and the flow diagrams appear simple.FISH AND SEAFOOD product. 8. It is essential that the diagram is accurate. because the hazard analysis and decisions regarding CCPs are based on these data.4.3. the qualitative and/or quantitative evaluation of the presence of hazards. However. 180 ● ● . it is essential that the details of each step are fully appreciated and recorded. then any pre-drawn HACCP plans must be reviewed once the lines/products are finalised.6 List all potential hazards associated with each step. features of equipment design.3. It may include details of the factory and equipment layout. Ideally. It is also important to include observation of production outside normal working hours such as shift patterns and weekend working. and any discrepancies between the flow diagram and normal practice must be recorded and the diagram amended accordingly. time/temperature data. or circumstances of any reclaim or rework activity. 8. 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.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. once determined. For example.
Note: it is important at this stage that no attempt is made to identify CCPs. Salmonella spp.1 Fish and seafood The term 'fish and seafood' includes an extremely varied group of products.6. The incidence of psychrotrophic. the hurdle effect. since this may interfere with the analysis.HACCP IN FISH AND SEAFOOD PRODUCT MANUFACTURE ● ● ● ● production or persistence of toxins. 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. Many of the microbiological hazards associated with fish and seafood products are derived from the raw materials. 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. One control measure may be relevant to several process steps. 181 . where a hazard is repeated. modified-atmosphere packaging (MAP) of chilled raw fish may provide conditions suitable for the growth of psychrotrophic C. or desirable. any food safety objectives or manufacturer’s food safety requirements. and viruses). 8. and there are an equally varied range of potential hazards associated with them. Therefore it is not possible. to generalise about expected hazards and the reader is once again referred to the appropriate product chapter for additional advice on specific hazards. or may originate from polluted water or from post-capture contamination (e. botulinum. Hazards specific to certain types of product are detailed in the appropriate chapters of this manual.g. there are particular hazards associated with contamination of shellfish by microorganisms from human sewage.).g. or poor control of batter mixes in frozen battered fish portions may allow growth of Staphylococcus aureus and production of enterotoxin. Pathogens may be part of the resident microflora of the living animal (e. For example. Hazards introduced during processing of fish products depend very much on the characteristics of the process. the number of consumers potentially exposed and their vulnerability. Vibrio spp. and the potential growth of Listeria monocytogenes on smoked fish. For example. The HACCP team should then determine what control measures exist that can be applied for each hazard. non-proteolytic Clostridium botulinum on fresh fish is also sufficiently high that its presence may be assumed.3.
Where possible. For example. and its use may not always be appropriate. it can be said that effective control measures are likely to include the following: • • • • • 8.2) to provide a logical.3. critical limits should be specified and validated for each CCP. it is not possible to generalise on likely CCPs.8 Careful selection of sources for raw materials Adequate temperature control Effective sanitation Food handler hygiene Prevention of cross-contamination Establish critical limits for each CCP Critical limits separate acceptable from unacceptable products. It is also essential that the HACCP team has access to sufficient technical data to determine the CCPs effectively. 8. However.FISH AND SEAFOOD 8. biological or sensory parameters. then the process must be modified to include a control measure. 8. structured approach to decision making.184.108.40.206 Determine CCPs The determination of CCPs in the HACCP system is facilitated by a decision tree (Figure 8. but for which no control exists at that step or any other. chemical. and the reader is referred to the appropriate product chapter.1 Fish and seafood Specific product chapters provide information on criteria that may be used to set critical limits.3. If a significant hazard has been identified at a step where control is required for safety. 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 . 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. More than one critical limit may be defined for a single step.1 Fish and seafood Again. Criteria used to set critical limits must be measurable and may include physical. However.3. it is necessary to specify both time and temperature for a thermal process. application of the decision tree should be flexible.
HACCP IN FISH AND SEAFOOD PRODUCT MANUFACTURE
• • • 8.3.9
Water activity (aw) values for dried fish pH values in fermented fish Microbiological quality of water in shellfish production areas Establish a monitoring system for each CCP
Monitoring involves planned measurement or observation of a CCP relative to its critical limits. Monitoring procedures for target levels must be able to detect loss of control of the CCP, and should provide this information with sufficient speed to allow adjustments to be made to the control of the process before the critical limits are violated. Monitoring at critical limits should be able to rapidly detect when the critical limit has been exceeded. Monitoring should therefore either be continuous, or carried out sufficiently frequently to ensure control at the CCP. Therefore, physical and chemical on-line measurements are usually preferred to lengthy microbiological testing. However, certain rapid methods, such as ATP assay by bioluminescence, may be useful for assessment of adequate cleaning, which could be a critical limit for some CCPs, for example pre-start-up hygiene. Persons engaged in monitoring activities must have sufficient knowledge, training and authority to act effectively on the basis of the data collected. These data should also be properly recorded. 8.3.10 Establish corrective actions
For each CCP in the HACCP plan, there must be specified corrective actions to be applied if the CCP is not under control. If monitoring indicates a deviation from the critical limit for a CCP, action must be taken that will bring it back under control. Actions taken should also include proper isolation of the affected product, repair of defective equipment, and an investigation into why the deviation occurred. A further set of corrective actions should relate to the target levels, if process drift is occurring. In this case, only repair of the process defect and investigation of the fault are required. All corrective actions should be properly recorded. 8.3.11 Establish verification procedures
Verification usually involves auditing and testing procedures. Auditing methods, procedures and tests should be used frequently enough to determine whether the HACCP system is being followed, and is effective at controlling the hazards. These may include random sampling and analysis, including microbiological testing. Although microbiological analysis is generally too slow for monitoring purposes, it can be of great value in verification, since many of the identified hazards are likely to be microbiological. In addition, reviews of HACCP records are important for verification purposes. These should confirm that CCPs are under control and should indicate the nature
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of any deviations and the actions that were taken in each case. It is also useful to review customer returns and complaints regularly. 8.3.12 Establish documentation and record keeping
Efficient and accurate record keeping is an essential element of a HACCP system. The procedures in the HACCP system should be documented. ● ● Determination of critical limits The completed HACCP plan
Examples of documented procedures include: ● ● The hazard analysis Determination of CCPs
Examples of recorded data include: ● ● ● Results of monitoring procedures Deviations from critical limits and corrective actions Records of certain verification activities, e.g. observations of monitoring activities, and calibration of equipment.
The degree of documentation required will depend partly on the size and complexity of the operation, but it is unlikely to be possible to demonstrate that an effective HACCP system is present without adequate documentation and records. The length of time that records are kept will be as per company policy, but should not be less than one year beyond the shelf life of the product. Three to five years is typical for many food companies. 8.4 Implementation and review of the HACCP plan
The completed plan can only be implemented successfully with the full support and co-operation of management and the workforce. Adequate training is essential and the responsibilities and tasks of the operating personnel at each CCP must be clearly defined.
HACCP IN FISH AND SEAFOOD PRODUCT MANUFACTURE
Fig. 8.2. CCP Decision Tree (Adapted from Codex Alimentarius Commission, 2003) Finally, it is essential that the HACCP plan be reviewed following any changes to the process, including changes to raw materials, processing conditions or equipment, packaging, cleaning procedures and any other factor that may have an effect on product safety. Even a small alteration to the product or process may invalidate the HACCP plan and introduce potential hazards. Therefore, the implications of any changes to the overall HACCP system must be fully
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considered and documented, and adjustments made to the procedures as necessary. Triggered reviews/audits should occur as a result of changes, whereas a scheduled review/audit should be annual, as a minimum. 8.5 References
Wareing P.W., Carnell A.C. HACCP – A Toolkit for Implementation. Leatherhead, Leatherhead Food International. 2007. Bernard D., Scott V. Hazard Analysis and Critical Control Point System: use in controlling microbiological hazards, in Food Microbiology: Fundamentals and Frontiers. Eds. Doyle M.P., Beuchat L.R. Washington D.C., ASM Press. 2007, 971-86. Su Y-C., Daeschel M.A. Sanitation and HACCP, in Surimi and Surimi Seafood. Ed. Park J.W. Boca Raton, CRC Press. 2005, 141-62. Huss H.H. Considerations in the application of the HACCP principles to seafood production, in Assessment and Management of Seafood Safety and Quality. Eds. Food and Agricultural Organisation, Huss H.H., Ababouch L., Gram L. Rome, FAO. 2004, 153-77. Chen H-C., Chang P.C-M. Frozen seafood safety and HACCP, in Handbook of Frozen Foods. Ed. Hui Y.H., Cornillon P., Legaretta I.G., Lim M.H., Murrell K.D., Nip WK. New York, Marcel Dekker. 2004, 339-63. Mortimore S., Mayes T. The effective implementation of HACCP systems in food processing, in Foodborne Pathogens: Hazards, Risk Analysis and Control. Eds. Blackburn C.deW., McClure P.J. Cambridge, Woodhead Publishing Ltd. 2002, 229-56. Mayes T., Mortimore C.A. Making the Most of HACCP: Learning from Other’s Experience. Cambridge, Woodhead Publishing. 2001. Mortimore S.E., Wallace C., Cassianos C. HACCP (Executive Briefing). London, Blackwell Science Ltd. 2001. Chartered Institute of Environmental Health. HACCP in Practice. London, Chadwick House Group Ltd. 2000. Jouve J.L. Good manufacturing practice, HACCP, and quality systems, in The Microbiological Safety and Quality of Food, Volume 2. Eds. Lund B.M., BairdParker T.C., Gould G.W. Gaithersburg, Aspen Publishers. 2000, 1627-55. Stevenson K.E., Bernard D.T. HACCP: A Systematic Approach to Food Safety. Washington D.C., Food Processors Institute. 1999. National Seafood HACCP Alliance. IQF cooked shrimp processing HACCP model, in HACCP User's Manual. Ed. Corlett D.A. Gaithersburg, Aspen Publishers. 1998, 454-73. Mortimore S., Wallace C. HACCP: A Practical Approach. Gaithersburg, Aspen Publishers. 1998.
HACCP IN FISH AND SEAFOOD PRODUCT MANUFACTURE Khandke S.S., Mayes T. HACCP implementation: a practical guide to the implementation of the HACCP plan. Food Control, 1998, 9 (2-3), 103-9. Forsythe S.J., Hayes P.R. Food Hygiene, Microbiology and HACCP. Gaithersburg. Aspen Publishers. 1998. Food and Agriculture Organisation. Food Quality and Safety Systems: A Training Manual on Food Hygiene and the Hazard Analysis and Critical Control Point (HACCP) System. Rome, FAO. 1998. US Food and Drug Administration. Fish and Fishery Products Hazards and Controls Guide. Rockville, Food Drug Administration. 1998. National Advisory Committee on Microbiological Criteria for Foods. Hazard Analysis and Critical Control Point Principles and Application Guidelines. 1997. Gardner L.A. Testing to fulfil HACCP (Hazard Analysis Critical Control Points) requirements: principles and examples. Journal of Dairy Science, 1997, 80 (12), 3453-7. Martin R.E., Collette R.L, Slavin J.W. Fish Inspection, Quality Control and HACCP: A Global Focus. Proceedings of a Conference, Arlington, May 1996. Lancaster P.A., Technomic Publishers. 1997. Codex Alimentarius Commission. Hazard Analysis Critical Control Point (HACCP) System and guidelines for its application, in Food Hygiene: Basic Texts. Ed. Codex Alimentarius Commission. Rome, FAO. 2003, 21-31. Garrett E.S., Jahncke M.l., Tennyson J.M. Microbiological hazards and emerging food safety issues associated with seafoods. Journal of Food Protection, 1997, 60 (11), 1409-15. National Seafood HACCP Alliance. HACCP: Hazard Analysis and Critical Control Point Training Curriculum. Raleigh, North Carolina State University. 1997. Dillon M., McEachern V. HACCP and quality assurance of seafood, in Fish Processing Technology. Ed. Hall G.M. London, Blackie. 1997, 224-48. Savage R.A. Hazard Analysis Critical Control Point: a review. Food Reviews International, 1995, 11 (4), 575-95. Pearson A.M., Dutson T.R. HACCP in Meat, Poultry and Fish Processing. Glasgow, Blackie. 1995. Huss H.H., Food and Agriculture Organisation. Assurance of Seafood Quality. FAO Fisheries Technical Paper No. 334. Rome, FAO. 1994. Bonnell A.D. Process control, in Quality Assurance in Seafood Processing: A Practical Guide. Ed. Bonnell A.D. London, Chapman & Hall. 1994, 39-53. Pierson M.D., Corlett D.A., Institute of Food Technologists. HACCP: Principles and Applications. New York, Van Nostrand Reinhold. 1992. Hudak-Roos M., Garrett E.S. Monitoring, corrective actions and record keeping in HACCP, in Quality Assurance in the Fish Industry: Proceedings of an International Conference, Copenhagen, August 1991. Eds. Huss H.H., Jakobsen M., Liston J. Amsterdam, Elsevier. 1992, 521-31.
1992.L. 1992. 1988. 14-9. Food Control. 15 (1/2). 3 (1). Oxford. Mayes T. International Journal of Food Microbiology. Blackwell Scientific Publications.H. International Commission on Microbiological Specifications for Foods. 33-44. Development and use of the HACCP concept in fish processing.HACCP IN FISH AND SEAFOOD PRODUCT MANUFACTURE Huss H.. Volume 4: Application of the Hazard Analysis Critical Control Point (HACCP) System to Ensure Microbiological Safety and Quality. Bryan F. Geneva. WHO. Simple users' guide to the hazard analysis critical control point concept for the control of food microbiological safety. Hazard Analysis Critical Control Point Evaluations: A Guide to Identifying Hazards and Assessing Risks Associated with Food Preparation and Storage. World Health Organisation. 1992. 188 . Microorganisms in Foods.
Procedures under the new legislation are based on risk assessment and management and follow a 'farm to fork' approach to food 189 . the existence of microbiological standards alone cannot protect consumer health. Microbiological standards have a useful role and help establish requirements for the microbiological safety and quality of food and raw materials. Hygiene is a general concept that covers a wide subject area. however. with the intention of introducing consistency and clarity throughout the food production chain from primary production. have been replaced by several linked measures on food safety rules and associated animal health controls. A number of standards are provided in food legislation. from structural conditions in the factory or process facility. and conditions of raw materials. eliminate or acceptably control pathogen contamination of food. 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. to personnel requirements. final product specifications. transport and delivery vehicles requirements. The new legislation was designed to establish conditions. and one that plays an important role in most countries’ food legislation. to sale or supply to the final consumer. It is generally considered that the principles of Good Manufacturing Practice (GMP) and application of Hazard Analysis Critical Control Point (HACCP) systems are of greater importance.1 Introduction Hygiene is an important aspect of ensuring food safety.9. EC FOOD HYGIENE LEGISLATION Eugenia Choi and Dr. under which food is produced to optimise public health and to prevent. A new package of European Commission (EC) hygiene measures became applicable on 1 January 2006 to update and consolidate the earlier 17 hygiene directives. Jenny Pfleger Leatherhead Food International Randalls Road Leatherhead Surrey KT22 7RY United Kingdom 9. including microbiological criteria.
FISH AND SEAFOOD safety with the inclusion of primary production in food hygiene legislation. 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 187.1992. Subsequently. A number of more detailed implementing and transitional measures have been adopted at EC level. 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. 41). 853/2004 of the European Parliament and of the Council laying down specific hygiene rules for food of animal origin Regulation (EC) No. Prescribed are detailed measures to ensure the safety and wholesomeness of food during preparation.15).1). 9. 7. European Communities L 268.B. storing. as last amended by Regulation (EC) No 806/2003. Regulation 853/2004 supplements Regulation 852/2004 in that it lays down specific requirements for food businesses dealing with foods of animal origin.1991. 2073/2005 on microbiological criteria for foodstuffs • The general hygiene requirements for all food business operators are laid down in Regulation 852/2004. transportation. p. handling and offering for sale or supply to the consumer.9. packaging. J.9. 24. 24. the following European Union (EU) hygiene regulations have applied: • • • Regulation (EC) No. p. as last amended by Regulation (EC) No 806/2003. N. 190 • • . manufacturing. 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. 852/2004 of the European Parliament and of the Council on the hygiene of foodstuffs Regulation (EC) No. distribution. p.2 Legislative Structure From 1 January 2006. Regulation 854/2004 relates to the organisation of official controls on products of animal origin. 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. processing. and sets out what those enforcing the provisions have to do.7.1991.
etc. for example. 19. 2073/2005. 191 . and equivalent legislation in Scotland. and binding in its entirety on all member states from the date of entry into force. national legislation in the form of a Statutory Instrument (S. As regulations. the legislation is directly applicable law. which will be covered later in this chapter. 852/2004 on the General Hygiene of Foodstuffs Food business operators must ensure that all stages of production. Primary products are defined as products of primary production including products of the soil. Wales and Northern Ireland. Additionally the requirements of EC Regulation 853/2004 must be complied with.I. The Food Hygiene (England) Regulations 2006 (S.3 Regulation (EC) No.I. food storage that is for private or domestic consumption.11) The EU hygiene regulations apply to all stages of food production including primary production. processing and distribution of food under their control satisfy the relevant hygiene requirements laid down in Regulation (EC) No. of stock farming. Wales and Northern Ireland).LEGISLATION • Council Directive 93/43/EEC of 14 June 1993 on the hygiene of foodstuffs (OJ L 175. penalties and powers of entry. e. 852/2004. revocation of existing implementing legislation.1993. is required to give effect to the EU regulations. and of hunting and fishing. 1 . right through to the implementation of procedures based on HACCP principles. particularly taking into account a number of factors ranging from ensuring food safety throughout the food chain to begin with primary production. This Regulation lays down general rules for food business operators on the hygiene of foodstuffs. and also if the producer supplies small amounts of primary product to the final consumer or local retail establishments supplying the final consumer. 9.7. There are some exemptions. 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.14. p. domestic preparation or handling. setting offences. 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. The national legislation in all four UK countries also applied the provisions of the EU Microbiological Criteria Regulation No. with primary production. Although the regulations have the force of law. as amended) came into force on 11 January 2006 (separate but similar national legislation also came into force on that day in Scotland. 2006 No. 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.g.) in England.
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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: 220.127.116.11 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.
18.104.22.168 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.
22.214.171.124 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.
126.96.36.199 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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188.8.131.52 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.
184.108.40.206 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.
220.127.116.11 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.
18.104.22.168 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. 22.214.171.124 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.
126.96.36.199 Chapter X Chapter X lays down provisions applicable to the wrapping and packaging of foodstuffs to avoid contamination of any form. 188.8.131.52 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). 184.108.40.206 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:
heading. in particular as a result of feeding on plankton containing toxins. to preserve or to improve organoleptic qualities and to ensure they are in a good state of vitality before wrapping or packing. 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. mincing or processing. to remove mud. Factory vessel means any vessel on board which fishery products undergo one or more of the following operations followed by wrapping or packaging and. after preparatory work such as bleeding. where necessary. skinning.LEGISLATION Fishery products means all seawater or freshwater animals (except for live bivalve molluscs. shelling. shucking. Fresh fishery products means unprocessed fishery products. followed by wrapping or packaging. heading. 197 . such as gutting. slime or sand. and all mammals. Freezer vessel means any vessel on board in which freezing of fishery products is carried out. whether whole or prepared. gutting and removal of fins and. live tunicates and live marine gastropods. Prepared fishery products means unprocessed fishery products that have undergone an operation affecting their anatomical wholeness. and chopping. 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. 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. where appropriate. live echinoderms. or in natural sites. slicing. parts and products of such animals. reptiles and frogs) whether wild or farmed and including all edible forms. Conditioning means the storage of live bivalve molluscs coming from class A production areas. slicing. if necessary: chilling or freezing: filleting. Bivalve molluscs means filter-feeding lamellibranch molluscs Marine biotoxins means poisonous substances accumulated by bivalve molluscs. filleting.
2. EB or WE.2 Fishery product requirements 9.1 Identification marking An identification mark needs to be applied following the requirements of Annex II.FISH AND SEAFOOD 9. EC. (ii) However.1. Extracts of the requirements of Annex II. In this case. EG.2. when a product's packaging and/or wrapping is removed or is further processed in another establishment.1. If an establishment manufactures both food to which Regulation 853/2004 applies and food to which it does not.4. 9. (iv) If the identification mark is applied in an establishment within the Community. For the UK. EF. the mark must show the approval number of the establishment. 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.4. the mark must be oval shaped and include the abbreviation CE.4. Section I of Regulation (EC) 853/2004 are given as follows: 9.1 Application of the identification mark (i) The identification mark must be applied before the product leaves the establishment of production. ES. please refer to Regulation (EC) 853/2004.4. (ii) The mark must indicate the name of the country in which the establishment is located and written out in full or shown as a two-letter code in accordance with the relevant ISO standard.2. 198 .2 Form of the identification mark (i) The mark must be legible and indelible. EÜ. the food business operator may apply the same identification mark to both types of food. for other codes. EK. the new mark must show the approval number of the establishment where this occurred. (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. EO. and the characters easily decipherable and be clearly displayed. EY. (iii) Additionally. EK. the code would be UK. Section I of Regulation (EC) 853/2004. a new mark must be applied to the product.
it is sufficient to apply the mark to the exterior of that package only.4. Therefore.2. its wrapping or the packaging. Part A and Chapter V apply to retail. This section does not apply to bivalve molluscs. an identification mark is not necessary if accompanying documentation contains the name of the country or its two-letter code. The requirements of this section accompany those in Regulation (EC) 852/2004. and fishery products carried in bulk. the wrapping or the packaging. and are intended for further handling. granulate and powdered products of animal origin carried in bulk. involved in primary production and such operations accompany the needs of Annex I of Regulation (EC) 852/2004 b) for other establishments including vessels. as amended. wrapping or packaging in another establishment.2.3 Method of marking (i) The mark is to be applied directly to the product. Parts A.e.LEGISLATION 9. Extracts of the requirements of Annex III. C and D. i. (iv) If products of animal origin are placed in a package destined for direct supply to the final consumer. With the exception of Chapters I and II. they would need to be obtained following Section VII on live bivalve molluscs. processing.4. it applies to those animals when not placed on the market live. It may also be a tag made of a resistant material affixed in such a way that it cannot be removed. including vessels. or be printed on a label affixed to the product. 9. the requirements accompany those of Annex II of Regulation (EC) 852/2004 199 . reference should be made to the actual Regulation. the colourings used must be EC authorised colours.2 Hygiene requirements for fishery products The regulation details specific hygiene requirements for fishery products. Chapter III. (iii) In the case of liquid. specifically relating to fishery products are given below.1. Chapter IV. the mark may be applied to the outer surface of the container or packaging. (ii) For products of animal origin that are placed in transport containers or large packages. (v) When the mark is applied directly to products of animal origin. a) requirements for establishments. for full requirements. echinoderms. Section VIII of Regulation 853/2004. tunicates and marine gastropods when placed on the market live. the approval number of the establishment and the abbreviation for the community where appropriate.
e. in that record keeping requirements stated by point 7 in Part A of Annex I of Regulation 852/2004 do not apply to operators involved in small-scale coastal fishing (as defined through Regulation (EC) 1198/2006) if the activity is under 24 hours. 2. or to handle or process them after harvesting. they also include (i) the transport and storage of fishery products that have not been substantially altered (including live fishery products.) 9. Chapter VII of Regulation 852/2004. and (b) related operations include any of the following operations if done on board fishing vessels: slaughter. from the place of production to the first establishment of destination. surfaces that contact fishery products need to be made from cleanable corrosion-resistant material..1 Chapter I . There is a derogation from point a. bleeding.2.2. Structural and equipment requirements refer to the following points 1. clean seawater may be used for the handling and washing of fishery products. A. if a vessel has water coming in that is used with fishery products. The definition of fishery products covers primary production as well as its associated operations. removing fins. refrigeration and wrapping.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. i. the production of ice used to chill fishery products and the rapid cooling of crustaceans and molluscs after their cooking. gutting. this supplements the requirements of Annex II.4. fishing and collection of live fishery products with a view to their being placed on the market.FISH AND SEAFOOD c) for the water supply. (a) primary production covers the farming. Requirements for all vessels Vessels should be made in a way that does not cause contamination with substances such as fuel and oil etc. heading. it must be placed somewhere that will not contaminate the water supply. Requirements for vessels designed and equipped to preserve fresh fishery products for more than 24 hours 200 . within fish farms on land) (ii) the transport of fishery products that have not been substantially altered (including live fishery products.
(v) Special equipment for disposing of waste/ inedible fishery products into the sea or a watertight tank and allocation of separate areas. (iii) The above requirements for holds in vessels designed and equipped to preserve fresh fishery products for more than 24 hours must be met. Storage areas for the finished products also need to be sufficiently large and cleanable. 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. 4. where necessary. and allow the monitoring and. any waste processing unit needs to have a separate hold to store the waste. and not more than 0 °C after 16 hours. where necessary. (iii) In vessels equipped for chilling fishery products in cooled clean seawater. recording of temperatures. 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. (ii) Refrigeration equipment needs to keep fishery products in storage holds at not more than -18 °C. 201 . 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. A temperature recording device must be present and placed where the temperature in the hold is the highest. Also. (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. tanks or containers for the storage of fishery products at the temperatures laid down in Chapter VII.LEGISLATION (i) Such vessels need to have holds. Holds and containers used for storing fishery products must allow its preservation under adequate hygiene conditions and. tanks must incorporate devices that allow a uniform temperature throughout the tanks. 3. (iv) There needs to be somewhere for storing packaging materials and this is to be separate from product preparation and processing areas.
and products washed thoroughly. Handlers may use spiked instruments to move large fish or fish that might injure them. 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. chilled and wrapped.2. b) Factory vessels on board which crustaceans and molluscs are cooked. B. landed as soon as possible.2. b) Fishery products are to be handled and stored in a way that prevents the flesh bruising. 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 . if this is not possible. c) Factory vessels that freeze fishery products must have equipment meeting the requirements for freezer vessels as stated above. the product must be kept at no more than -9 °C. other than those kept alive.2 (i) Unloading and landing equipment that comes into contact with fishery products needs to be made of material that is easily cleanable. Livers and roes for human consumption must be preserved under ice (temperature to approach that of melting ice) or be frozen.4. must be chilled or. 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. d) Any gutting or heading of fish on board is to be done hygienically and quickly after capture. (ii) In the case of food business operators responsible for auction and wholesale markets etc. c) Fishery products. where fishery products are displayed for sale. do not have to meet the above requirements if no other form of handling or processing takes place on board such vessels. e) In cases where whole fish intended for canning is frozen in brine.Requirements during and after landing 9. The brine must not be a source of contamination. Chapter II . Parts that may present a danger to public health are to be removed quickly and kept away from products for human consumption.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. provided no damage occurs to the flesh.
Fillets and slices must then be removed from work tables as soon as possible after preparation.LEGISLATION consumption. They should be wrapped. packaged and chilled as quickly as possible after preparation. including vessels. Requirements for mechanically separated fishery products (i) Raw materials used must satisfy the following 203 . there are requirements that such premises should be well lit and vehicles emitting fumes should not be able to access the premises. Gutting should ideally be done quickly once products have been caught or landed and afterwards be washed thoroughly.3 Chapter III . C. 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. Chapter I.4. (iii) Whole and gutted fresh fishery products can be transported and stored in cooled water on board vessels.2. and where necessary. (ii) Heading and gutting must be carried out hygienically. handling fishery products A. Packaged fresh fishery products must be chilled to a temperature approaching that of melting ice. part I. Requirements for fresh fishery products (i) Where chilled or unpackaged products are not distributed. 854/2004. B. (iv) Filleting and cutting is to be carried out in a way that avoids contamination or spoilage of fillets or slices. (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. points 1 and 2. prepared or processed immediately after arrival at an establishment on land. dispatched.Requirements for establishments. 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. they must be stored under ice in appropriate facilities. 9.2. C. Additionally. 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.
(iii) sprat. (b) fishery products from the following species.FISH AND SEAFOOD Only whole fish and bones after filleting may be used to produce mechanically separated fishery products. Requirements concerning parasites (i) The fishery products below must be frozen at no more than .Requirements for certain processed fishery products A. 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. must accompany fishery products referred to above when placed on the market. and whole fish must be gutted and washed beforehand. they must be cooled rapidly.4. raw materials must have no guts.2. Requirements for cooking of crustaceans and molluscs After crustaceans and molluscs are cooked. stating the type of process they have undergone. (ii) mackerel. or a stabilising treatment. except when supplied to the final consumer.20 °C in all parts of the product for at least 24 hours (this treatment must be applied to the raw product or the finished product): (a) fishery products to be consumed raw or almost raw. Once produced. (ii) The manufacturing process must satisfy the following Mechanical separation is to be carried out immediately after filleting. If no other method of preservation is used. (iv) (wild) Atlantic and Pacific salmon. mechanically separated fishery products must be frozen as quickly as possible or incorporated into a product intended for freezing. and (c) marinated and/or salted fishery products. (iii) A document from the manufacturer. 9. cooling must continue until a temperature 204 . if the processing is insufficient to destroy nematode larvae.2. (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. D.4 Chapter IV .
centrifugation. and after this. B. C. B. and the raw material is processed within 36 hours after loading. Histamine Histamine limits must not be exceeded (limits are set in Regulation (EC) No. Organoleptic properties of fishery products An organoleptic examination of fishery products must be carried out to ensure that fishery products comply with any freshness criteria (such criteria have yet to be set). 2074/2005. 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. 9. raw materials need to be chilled as soon as possible and should meet the temperature requirements in Chapter VII.2. However.4. 205 . 2074/2005). 2073/2005). processing. cooked products are to be frozen or chilled to the temperatures stated in Chapter VII. the following conditions need to be met: A. separation.5 Chapter V . pressing. 2073/2005. 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. refining and purification steps before being placed on the market for the final consumer.2.LEGISLATION approaching that of melting ice is reached. (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. 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.Health standards for fishery products In addition to compliance with microbiological criteria laid down in Regulation (EC) No. Shelling/shucking is to be done hygienically. Requirements for fish oil intended for human consumption (i) In relation to the raw materials used in their preparation and their transport.
6 Chapter VI .Storage of fishery products (i) Fresh fishery products. (ii) Frozen blocks prepared onboard vessels must be adequately wrapped before landing. 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.2. may only be placed on the market in wrapped/packaged form and must be appropriately labeled to provide information to the consumer on preparation/cooking methods and on the risk related to the presence of substances with adverse gastrointestinal effects.2. and when fishery products are wrapped on board fishing vessels. Toxins harmful to human health Fishery products derived from poisonous fish of the following families must not be placed on the market: Tetraodontidae.4. The scientific name must accompany the common name on the label. 206 . prepared and processed fishery products belonging to the family Gempylidae.2.2.FISH AND SEAFOOD D. 9. it should be kept at no more than -9 °C. In the case of whole frozen fish in brine intended for the manufacture of canned food. Parasites Fishery products are to be visually examined so that visible parasites are identified before being placed on the market. and cooked and chilled products from crustaceans and molluscs are to be kept at a temperature close to that of melting ice. 9. Diodontidae and Canthigasteridae.4. Fresh. Fishery products contaminated with parasites may not be placed on the market for human consumption. in particular Ruvettus pretiosus and Lepidocybium flavobrunneum. Molidae. thawed unprocessed fishery products.7 Chapter VII . E. (ii) Frozen fishery products must be kept at a temperature of not more than -18 °C in all parts of the product.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. The requirements for histamine and parasites do not apply to whole fishery products being used directly for preparing fish oil intended for human consumption.
The required temperature for whole frozen fish in brine intended for the manufacture of canned food during transport is not stated.). species belonging to the Gadidae family. 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.8 Chapter VIII . provided that the journey is short and the competent authority allows this. This covers Total volatile basic nitrogen (TVBN) limits and how to determine them.3 Other information: Rules relating to visual inspections for the detection of parasites in fishery products are stated in Annex II (obligations on the competent authorities) of Regulation (EC) No. 2074/2005.4. Sebastichthys capensis. (c) 35 mg of nitrogen/100 g of flesh for the species: Salmo salar. melt water must not remain in contact with the products. Note: (i) Frozen fishery products do not need to be kept at a temperature of -18 °C if they are being transported from a cold store to an approved establishment to be thawed on arrival for the purposes of preparation and/or processing. 9. (b) 30 mg of nitrogen/100 g of flesh for the species belonging to the Pleuronectidae family (with the exception of halibut: Hippoglossus spp. Helicolenus dactylopterus. (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 .4. The temperature of frozen fishery products must not fluctuate above -15°C. species belonging to the Merlucciidae family.LEGISLATION (iii) Fishery products kept alive are to be kept at a temperature which will not adversely affect food safety or its viability.Transport of fishery products The above points for Chapter VII apply in relation to transport. 9.2. (ii) If fishery products are kept under ice.2.2.
Member States may set limits at a higher level for certain species pending the establishment of specific Community legislation. taken from at least three different points and mixed together by grinding. tunicates and marine gastropods. Apart from the provisions on purification. The information required in the registration document is given in point 4 and includes the health status of the production area. 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. Extracts of the requirements of Annex III.FISH AND SEAFOOD Regulation (EC) No 853/2004.4. and point 3 of Chapter VII applies to retail. (b) and (c) of Part B(1) of that Chapter. (b) In the case of other operations. they supplement the requirements of Annex I to that Regulation. The sample must consist of about 100 g of flesh. Chapter IX applies to pectinidae harvested outside those areas. VIII and IX.1 Chapter I . specifically relating to bivalve molluscs are given below. as amended.3 The regulation details specific hygiene requirements for bivalve molluscs. A food business operator can only accept batch of such bivalve molluscs provided correct documentary requirements are presented.3.4. The copy of 208 . where the raw material complies with points (a). please refer directly to the regulation. for full requirements. however. this Section also applies to live echinoderms. Chapters V. 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. Live bivalve mollusc requirements - 9. they supplement the requirements of Annex II to that Regulation. shellfish species etc. 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. reference should be made to the actual regulation. Section VII of Regulation 853/2004. 9.General requirements for the placing on the market of live bivalve molluscs Live bivalve molluscs can only be put on the market for retail sale if done through a dispatch centre. VI.
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. collection from: .LEGISLATION document must be kept for at least 12 months post dispatch or receipt or longer as stated by the competent authority. Other requirements such as conditions for relaying in order to allow optimal conditions for purification and for permanent record keeping are also specified. 209 . 9. 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. (ii) Provided the requirements of chapter V are met. C. processing or relaying.4. poles or other fixed means are to identify the boundaries of the sites.3. 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.Hygiene requirements for the production and harvesting of live bivalve molluscs A. Transport needs to allow sufficient drainage for best survival conditions.Class C means relaying over a long time before it can be placed on the market For bivalve molluscs not subject to Class B or C treatments.Class A means they can be placed directly on the market . B. 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 . Requirements for production areas (i) The competent authority classes (classed A.2 Chapter II .
4.Structural requirements for dispatch and purification centres Premises on land should not be subject to flooding by tides etc.FISH AND SEAFOOD 9. requirements if there are many batches of live bivalve molluscs. 160 micrograms of okadaic acid equivalents per kilogram.5 Chapter V . 1 milligram of yessotoxin equivalent per kilogram. 210 .Hygiene requirements for purification and dispatch centres A. 20 milligrams of domoic acid per kilogram (iii) okadaic acid.3. Requirements for purification centres There are requirements for live bivalve molluscs to be washed.4.4 Chapter IV . B. 9. (ii) amnesic shellfish poison (ASP).3 Chapter III . containers and a label to identify purification. 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. and there are requirements for tanks and water storage containers. the purification system to allow them to resume /maintain filter-feeding activity. Before dispatch. (iv) yessotoxins. the levels of marine biotoxins should not exceed the following limits.Health standards for live bivalve molluscs In addition to microbiological criteria and organoleptic characteristic requirements.4. Requirements for dispatch centres Handling should not result in contamination to any degree. dinophysistoxins and pectenotoxins together. 800 micrograms per kilogram. for: (i) paralytic shellfish poison (PSP).3.3.
Also. the label.Specific requirements for pectinidae harvested outside classified production areas Requirements for harvesting and handling following harvesting and health standards for live bivalve molluscs must be met. 9.8 Chapter VIII . The requirements for production areas as stated earlier apply to pectinidae and these can only be placed on the market through a fish auction.3. 160 micrograms of azaspiracid equivalents per kilogram.Wrapping and packaging of live bivalve molluscs (i) Oysters must be wrapped or packaged with the concave shell downwards.4. Finally.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. (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. 211 .9 Chapter IX .3.2. (ii) Once the live bivalve molluscs have been packed for retail sale and have left the dispatch centre.Identification marking and labelling In addition to general identification marking (contained in 9.4. they cannot be re-immersed in or sprayed with water. 9.4. the species of bivalve mollusc and date of packaging (at least the date and month) are required. Registration document requirements also apply. by way of derogation from directive 2000/13/EC. (Note: testing methods for detecting marine biotoxins are detailed in Annex III of Regulation (EC) 2074/2005) 9. 9. dispatch centre or processing establishment.LEGISLATION (v) azaspiracids.6 Chapter VI .1).4.3. including the identification mark.3.7 Chapter VII .4. Requirements of Chapters III and IV are to be complied with. the date of minimum durability may be replaced by the words ‘these animals must be alive when sold’. must be waterproof.
(ii) fishery products not complying with the requirement concerning biotoxins. they derive from: (i) poisonous fish.1 Regulation (EC) 854/2004 Annex III Chapter III states that fishery products are to be declared unfit for human consumption if: organoleptic. 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. or - - 212 .FISH AND SEAFOOD 9. Therefore the health standards for fishery products as previously stated in Annex III. This refers to official controls or production and placing on the market of fishery products. In this regulation. tunicates or marine gastropods containing marine biotoxins in total quantities exceeding the limits referred to in Regulation (EC) No 853/2004. general principles for official controls in respect of all products of animal origin falling within the scope of the regulation are given. The requirements refer to hygienic conditions and regular inspections both on land and at port to ensure compliance of temperature and other requirements. Section VIII. 854/2004 of the European Parliament and of the Council Laying Down Specific Rules for the Organisation of Official Controls on Products of Animal Origin Intended for Human Consumption Regulation (EC) 854/2004 gives requirements for official controls on products of animal origin and states requirements for those enforcing the provisions.5 Regulation (EC) No. echinoderms. Fishery products in particular must comply with the requirements of Annex III of the regulation. physical or microbiological checks or checks for parasites have shown that they are not in compliance with the relevant Community legislation. or (iii) bivalve molluscs. chemical. Chapter V of Regulation 853/2004 apply. The official controls include audits of good hygiene practices and hazard analysis and critical control point (HACCP)-based procedures.5. 9. they contain in their edible parts contaminants or residues in excess of the limits laid down in Community legislation or at levels where the calculated dietary intake would exceed the acceptable daily or weekly intake for humans.
in 90% of the samples. 213 . A summary is given as follows: 9.2 This states official controls for the production and placing on the market of live bivalve molluscs. three dilutions MPN test specified in ISO 16649-3. Monitoring of classified relaying and production areas This is to ensure there is no malpractice.600 E.5. coli per 100 g of flesh and intravalvular liquid. coli per 100 g of flesh and intravalvular liquid. Requirements for the competent authority are stated in relation to the classification or a production or relaying area is stated. 4.000 E. that microbiological criteria and chemical contaminants amongst other requirements are met. 000 E. live bivalve molluscs must not exceed 46.5. The reference method for this analysis is the five-tube.Official controls concerning live bivalve molluscs from classified production areas A. live tunicates and live marine gastropods. Live bivalve molluscs from class A areas may be collected for direct human consumption provided health standards are met. Live bivalve molluscs from Class B areas must not exceed. Regulation (EC) 854/2004. chapter V is reinforced. coli per 100 g of flesh and intravalvular liquid. The reference method for this analysis is the five-tube.2. Live bivalve molluscs from Class C areas must not exceed 46.1 Chapter II . three dilutions Most Probable Number (MPN) test specified in ISO 16649-3. live echinoderms. chapter II on hygiene requirements for the production and harvesting of live bivalve molluscs in Regulation (EC) 853/2004.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. as previously stated. section VII. Classification of production and relaying areas: This section reinforces that specified in Annex III. - - - - B. Annex II 9. Sampling plans and frequency are required. Also. In the remaining 10 % of samples.
2073/2005. F. 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. Additional monitoring requirements This concerns the competent authority in relation to monitoring of classified production areas where bivalve mollusc harvesting is forbidden and of relaying and production zones to ensure there is no malpractice etc. 9.5. boundaries or class of a production area or its closure.5. E. which has applied since 1 January 2006. the Member States may permit the import of bivalve molluscs and fishery products from the countries listed respectively in Annex I and Annex II to regulation (EC) 2076/2005. Recording and exchange of information Requirements are stated for competent authorities in relation to establishing and maintaining a current list of approved production and relaying areas and their role to inform interested parties of any changes of location. opening or closing a production area.6 Regulation (EC) No. 9.3 Imports By way of derogation from Article 11(1) of Regulation (EC) No 854/2004. 2073/2005 on Microbiological Criteria for Foodstuffs Regulation (EC) No.FISH AND SEAFOOD C.2 Chapter III .2. dispatch centres and processing establishments in order to confirm that health standards and other specific requirements are met. 214 .Official controls concerning pectinidae harvested outside classified production areas These are to be carried out in fish auctions. the competent authority is to close the production area. establishes microbiological criteria for a range of foods. 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. 9.
6. and the legislation applies to all businesses involved in food production and handling. with chapter 2.1 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. Two kinds of criteria have been established: food safety criteria.I. The relevant criteria are outlined in Table 9. ensuring that foods being placed on the market do not pose a risk to human health.2 Process hygiene criteria Chapter 2 focuses on process hygiene criteria. per unit(s) of mass. The criteria are outlined in Table 9.II. volume. a batch of foodstuffs or a process.4 referring to fishery products.LEGISLATION The aim of this legislation is to complement food hygiene requirements. 215 . 9. based on the absence. The definition of ‘microbiological criterion’ means a criterion defining the acceptability of a product. presence or number of microorganisms. 9. and/or on the quantity of their toxins or metabolites. applying to products placed on the market and process hygiene criteria that are applied during the manufacturing process.6. area or batch.
16 Cooked crustaceans and molluscan shellfish Salmonella EN/ISO 6579 Products placed on the market during their shelf life . monocytogenes 5 0 Absence in 25 g (7) FISH AND SEAFOOD 216 5 0 100 cfu/g 5 0 Absence in 25 g 1.3 Ready-to-eat foods unable to support the growth of L. other than those intended for infants and for special medical purposes(4) (8) L.1 Ready-to eat foods intended for infants and ready-to-eat foods for special medical purposes (4) 5 0 100 cfu/g (5) EN/ISO 11290-2 (6) L. monocytogenes.2 Ready-to-eat foods able to support the growth of L. other than those intended for infants and for special medical purposes EN/ISO 11290-1 Products placed on the market during their shelf life Before the food has left the immediate control of the food business operator. monocytogenes 1. who has produced it EN/ISO 11290-2 (6) Products placed on the market during their shelf life 1. monocytogenes.I Food safety criteria Sampling plan(1) n 10 5 Absence in 25 g EN/ISO 11290-1 c m M Products placed on the market during their shelf life Limit(2) Analytical reference method(3) Stage where the criterion applies Food category Microorganisms 1. monocytogenes L.TABLE 9.
27 Fishery products which have undergone enzyme maturation treatment in brine.17 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 .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. 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.
— — bottled or packed waters. products with pH ≤ 5. c = number of sample units giving values between m and M For point 1.0 and aw ≤ 0. AOAC Internat.94. Malle P. — bread. products heat treated in their final package). m = M.FISH AND SEAFOOD 1 2 3 4 n = number of units comprising the sample. Coryfenidae. Single samples may be taken at retail level.1 . that the product will not exceed the limit of 100 cfu/g throughout the shelf life. that the product will not exceed the limit 100 cfu/g throughout the shelf life. 5 This criterion shall apply if the manufacturer is able to demonstrate.25.92. coli is used here as an indicator of faecal contamination. when he is not able to demonstrate. Dervin C. Valle M. Products with pH ≤ 4. Bouquelet S. honey and confectionery. Clupeidae. cider. In such a case the presumption laid down in Article 14(6) of Regulation (EC) No 178/2002. subject to scientific justification. Pomatomidae. according to which the whole batch is to be deemed unsafe. The most recent edition of the standard shall be used. 43-9. Duflos G. — fresh. 2. beer. products with a shelf-life of less than five days shall be automatically considered to belong to this category.. Malle P. uncut and unprocessed vegetables and fruits. Other categories of products can also belong to this category. shall not apply. Particularly fish species of the families: Scombridae. excluding sprouted seeds. 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. This criterion shall apply to products before they have left the immediate control of the producing food business operator. Bouquelet S. 1996. wine.. 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. Assay of biogenic amines involved in fish decomposition. References: 1. monocytogenes. E.. Engraulidae. 79. sugar. to the satisfaction of the competent authority.4 or aw ≤ 0. biscuits and similar products. Scombresosidae.. to the satisfaction of the competent authority. A pooled sample comprising a minimum of 10 individual animals. spirits and similar products. Relevance of matrix effect in determination of biogenic amines in 6 7 8 15 16 17 18 19 218 . soft drinks. live bivalve molluscs. 1 ml of inoculum is plated on a Petri dish of 140 mm diameter or on three Petri dishes of 90 mm diameter.. including cocoa and chocolate products. J. when recontamination is not possible after this treatment (for example.1.
if any of the values are > the limit. J. — unsatisfactory. where the limit refers to a pooled sample.I The limits given refer to each sample unit tested. AOAC Internat. if the presence of the bacterium is detected in any of the sample units. — unsatisfactory. L. excluding live bivalve molluscs and live echinoderms. a maximum of c/n values observed are between m and M 3. The test results demonstrate the microbiological quality of the batch tested(1). monocytogenes in ready-to-eat foods able to support the growth of L. if the following requirements are fulfilled: 1. 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. 82. 1097-101. — unsatisfactory. monocytogenes in ready-to-eat foods intended for infants and for special medical purposes: — satisfactory. tunicates and gastropods in relation to testing E. 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. (1) 219 . L. Salmonella in different food categories: — satisfactory. monocytogenes in other ready-to-eat foods: — satisfactory. the mean value observed is ≤ m 2. if all the values observed indicate the absence of the bacterium. Histamine in fishery products from fish species associated with a high amount of histidine: — satisfactory. if all the values observed indicate the absence of the bacterium. L.LEGISLATION plaice (Pleuronectes platessa) and whiting (Merlangus merlangus. coli. Interpretation of the test results relating to Table 9. no values observed exceed the limit of M. 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. — unsatisfactory. if all the values observed indicate the absence of the bacterium. if the presence of the bacterium is detected in any of the sample units. 1999. if all the values observed are ≤ the limit. — unsatisfactory. if the presence of the bacterium is detected in any of the sample units.
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 .II Process hygiene criteria Limit(2) Analytical reference method(3) M 10 cfu/g ISO TS 16649-3 End of the manufacturing process End of the manufacturing process Improvements in production hygiene Stage where the criterion applies Action in case of unsatisfactory results Food category Microorganisms c 2 1 cfu/g m Sampling plan(1) n 2.4.TABLE 9.1 Shelled and E.
7. 2006 No. if a maximum of c/n values are between m and M. and the rest of the values observed are ≤ m. which overrides the 8 °C requirement. — acceptable. The defendant may be required to produce well-founded scientific proof to support his claims. had not been 221 .I. if all the values observed are ≤ m.1 In the U. — unsatisfactory.7 Food Hygiene (England) Regulations 2006. The regulations prescribe a chilled food holding temperature of 8 °C or less. 2006 No. if one or more of the values observed are > M or more than c/n values are between m and M. the defendant will need to prove that the food was on service or display. The regulations provide for defences in relation to upward variations of the 8 °C temperature. 14 details temperature control requirements for foods in general. and particularly that perishable foodstuffs must not be kept at above the maximum recommended storage temperature.II The limits given refer to each sample unit tested. 2 Interpretation of the test results relating to Table 9. coli in shelled and shucked products of cooked crustaceans and molluscan shellfish: — satisfactory. 9. 14 (Hygiene requirements specific to the U.K) Temperature control requirements 9. and the rest of the values observed are ≤ m. — acceptable. Coagulase-positive staphylococci in shelled and cooked crustaceans and molluscan shellfish: — satisfactory. — unsatisfactory.LEGISLATION 1 n = number of units comprising the sample. but there is also a general requirement that foods must not be kept at temperatures that would result in a risk to health. The test results demonstrate the microbiological quality of the process tested. For example. S.I. if one or more of the values observed are > M or more than c/n values are between m and M. if a maximum of c/n values are between m and M. tolerance periods for chill-holding of foods and hot-holding variations.K. c = number of sample units giving values between m and M The most recent edition of the standard shall be used. 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. if all the values observed are ≤ m. S. Schedule 4 of the Food Hygiene (England) Regulations 2006. with chill holding tolerance periods. E.
to or from premises (including vehicles) at which the food was to be kept at or below 8 °C or the recommended temperature.food.FISH AND SEAFOOD previously put on display at more than 8 °C and had been kept there for less than four hours. It should be noted that the temperature control requirements as detailed in Schedule 4 of the Food Hygiene (England) Regulations 2006 (S.S. was kept at above 8 °C or the recommended temperature for an unavoidable reason. Recommended temperature: a specified temperature that has been recommended in accordance with a food business responsible for manufacturing. the period for which it can be expected to remain fit for sale when kept in a manner consistent with food safety. The permitted reasons are given below: to facilitate handling during and after processing or preparation the defrosting of equipment. or.I. S. 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. 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. preparing or processing the food recommending that it be kept at or below a specified temperature between 8 ºC and ambient temperatures. 2006 No. Schedule 4 of the Food Hygiene (England) Regulations 2006 contains several definitions. including: Shelf life: where the minimum durability or 'use by' indication is required according to Regulation 20 or 21 of the Food Labelling Regulations 1996 (form of indication of minimum durability and form of indication of ‘use-by date’) the period up to and including that date. or temporary breakdown of equipment For Scotland there are separate provisions to include requirements to hold food under refrigeration or in a cool ventilated place.uk/foodindustry/guidancenotes/hygguid/fhlguidance/ 222 .I.gov. 3). 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.K. 9. and was kept at above 8 °C or the recommended temperature.8 Guidance In the U. such as that below. For other food. 2006 No. Alternatively. for a limited period consistent with food safety. it would need to be proved that the food was being transferred to or from a vehicle used for the activities of a food business.
852/2004 of the European Parliament and of the Council on the hygiene of foodstuffs (OJ No. 2076/2005. 25. 1019/2008 and as read with Regulation 2073/2005. these are not be discussed here.6. Regulation (EC) No. and Regulation (EEC) 1536/92 laying down common marketing standards for preserved tuna and bonito. as amended. 2074/2005 and Regulation (EC) No.4. 22) as amended by Regulation (EC) Nos. 62) as amended by Regulation (EC) No. 854/2004 is now set out in a Corrigendum (OJ No. Commission Regulation (EC) No. laying down common marketing standards for preserved sardines. Regulation (EC) No. 2075/2005 and Regulation (EC) No.4. 223 . 25. 55). The revised text of Regulation (EC) No. 2076/2005. 30.2004.LEGISLATION 9. 3) as amended by Regulation (EC) No. L139. 2073/2005 on microbiological criteria for foodstuffs (OJ No. L226.2004. Regulation (EC) No. 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. 1688/2005.2004. 83) as amended by Regulation Nos 882/2004.2006. 1. 1). namely. L283.6. Regulation (EC) No. 853/2004 is now set out in a Corrigendum (OJ No.12.10 Further Reading Regulation (EC) No. 1441/2007.2005. 1791/2006 and 1021/2008 and as read with EC Directive 2004/41. 2074/2005. 30. 2074/2005. L139. 30. The revised text of Regulation (EC) No. Regulation (EEC) 2136/89. L139. The revised text of Regulation (EC) No. 25. L226. 14. 853/2004 of the European Parliament and of the Council laying down specific hygiene rules for food of animal origin (OJ No. 22. L226. 1791/2006 and 1020/2008 and as read with EC Directive 2004/41. 2076/2005. 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. 9. 2076/2005. 1663/2006. 2074/2005. L338.2004. as read with the corrigenda at OJ No. 206). 1662/2006.6. Regulation (EC) No. 852/2004 is now set out in a Corrigendum (OJ No.10.2004.4.2004.
22.2005. 21. 22) Commission regulation (EC) No 2065/2001 laying down detailed rules for the application of Council Regulation (EC) No 104/2000 as regards informing consumers about fishery and aquaculture products as amended by Commission Regulation (EC) No 1792/2006 of 23 October 2006 (OJ L 278. 27 Commission Regulation (EC) No 2076/2005 laying down transitional arrangements for the implementation of Regulations (EC) No 853/2004.2001.12. 83) Regulation (EC) No 104/2000 on the common organisation of the markets in fishery and aquaculture products (OJ L 17.2000. as amended by Commission Regulation (EC) No 1022/2008 OJ L 338. 6) 224 .2005.1. derogating from Regulation (EC) No 852/2004 of the European Parliament and of the Council and amending Regulations (EC) No 853/2004 and (EC) No 854/2004.10.12. 22. 23.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. (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 Aeromonas spp.3 Temperature Aeromonas grows at temperatures of 2 .5.4 Heat resistance Aeromonads are sensitive to moderately high temperatures. 10. no growth is observed (1).0 .5 µm 10. Aeromonas has the potential to grow over a pH range of ca 4 . The optimum growth temperature is 28 °C (2). The minimum pH value for growth is thought to be pH 5. and 2. Some death will occur at this pH if the temperature is 4 °C. Morphology Gram-negative rods.2 Oxygen requirements Facultative anaerobe.1.3 minutes. a 10 to 1. respectively (1). 0. At 5 °C. and D-values at 45 and 51 °C have been reported as 29.10. under optimal incubation temperatures.1.000-fold increase in number is observed after one week in food systems (1). At pH 4.5 pH Under otherwise optimal conditions.5 minutes.10 (3). 10.1.45 °C (1).3 . PATHOGEN PROFILES 10.1. 10.3.5.1 10.1.0 x 1.1. 225 .
30 minutes. but Group II strains are psychrotrophic.5 . and is sensitive to > 4. 10. but the spores of this organism are more so. 10. botulinum are not particularly heat resistant. Group I will not grow at temperatures of 10 ºC or less. but the minimum temperatures required to inhibit Groups I (proteolytic group) and II (non-proteolytic group) are different.4 Heat resistance The vegetative cells of C. but the interior of the food may do so.2.FISH AND SEAFOOD 10. It is also important to note that the inclusion of oxygen as a packaging gas cannot ensure that growth of C.7 . F are the types that are responsible for most cases of human botulism (5. botulinum is prevented. 226 . at 85 ºC for 5 minutes. Thus an aerobically packed product may not support the growth of the organism on the surface. 10. All C. being capable of slow growth and toxin production at low temperatures . spore-forming rods.2.1 Morphology Gram-positive. 10.3 Temperature All strains of C.5% NaCl (4).2. There are four main groupings of the organism. and groups I and II are those responsible for the botulism cases. botulinum is an obligate anaerobe. 10.2 Clostridium botulinum Seven different types of C.even as low as 3 ºC (7).6 Aw/Sodium chloride Growth of Aeromonas is optimal in the presence of 1 .2% sodium chloride (NaCl) (1). Types A. botulinum are known. A to G. 0.2 Oxygen requirements Although C.4 x 1. or at 90 ºC for a few seconds. which may be inactivated by heating at 80 ºC for 20 .1. botulinum types produce heat labile toxins. types B and F can be either. 6). many foods that are not obviously ‘anaerobic’ can provide adequate conditions for growth.0 µm. B. E and. All type A strains are proteolytic and type E strains are usually non-proteolytic.2.22. botulinum grow reasonably well in the temperature range of 20 45 ºC.2. forming at least seven different toxins. to a lesser extent.
1 Clostridium perfringens Morphology Gram-positive spore-forming rods. and 5% (w/w) NaCl is necessary to prevent growth of Group II organisms. but under optimum growth conditions 10% (w/w) NaCl is required to prevent growth of Group I. These concentrations correspond to limiting aw of 0. It will not. 11)). perfringens .10. where oxygen levels are reduced.97) (10.7 Characteristics of C. botulinum spores The most heat resistant spores of Group I C.6 and pH 5. However. they may survive mild heat treatments (70 . therefore. These values have been established under carefully controlled laboratory conditions. The organism will grow well in the centre of meat or poultry dishes. Note also that if humectants other than salt are used. parcooked meals with pH value > 5.5 pH The minimum pH for the growth of proteolytic and non-proteolytic strains is pH 4. 10. 227 .g.3 10.85 ºC) and their ability to grow at refrigeration temperatures necessitates their control in foods capable of supporting their growth (e.3.97 for Group II (5). 10.is an obligate anaerobe.0. The spores of Group II (non-proteolytic/psychotrophic) strains are less heat resistant than Group I strains. particularly by cooking. botulinum are produced by type A and proteolytic B strains (D121ºC=ca 0.3 .6 Aw/ Sodium chloride The minimum water activity (aw) for growth of C. 9).0 and aw > 0.21 minutes) (9). pH and temperature.9 x 2. In commercial situations.94 for Group I and 0.0 . the minimum aw for growth may be lower. 10.like other clostridia .2.1.2 Oxygen requirements C. vacuum packed. respectively (8.2. 10.PATHOGEN PROFILES 10.3. grow on the surface of foods unless they are vacuum or gas-packed. 0. botulinum depends on solute.0 µm. safety margins must be introduced to allow for process variability.2.
perfringens can vary quite considerably in their heat resistance. 13). depending on strain and other conditions. perfringens is not tolerant of low water activities. Recorded heat 228 . perfringens in vacuum packed beef (18). very limited growth may occur at pH values over the range pH ≤ 5 and ≥ 8. will survive greater extremes of pH (and aw) (12. Salt concentrations of 6 . 15. In addition.10 ºC. Under otherwise ideal conditions.FISH AND SEAFOOD 10. perfringens appears to be 0.6 Aw/Sodium chloride C. the enterotoxin is not heat resistant .5 minutes (17). 13).50 ºC. 10.5 pH C. While some growth can occur at 50 ºC. although prior growth at high temperatures or the presence of fat in a food will result in increased heat resistance. Spores. pH.97 (glycerol and sucrose respectively) depending on the solute used to control aw (13.3.4 Heat resistance Exposure to a temperature of 60 ºC or more will result in the death of vegetative cells of C.3. perfringens strains.93 and 0.3 Temperature The most significant characteristic of C. Its optimum temperature for growth is 43 . The D-value for C. perfringens spores The spores of C.3. however. 16). the limits for aw are affected by temperature.3.it is destroyed by heating at 60 ºC for 10 minutes (14. vegetative cells die rapidly (12). perfringens has the potential ability to grow within the temperature range 15 . perfringens. The lowest aw recorded to support the growth of C. death of the vegetative cells of this organism usually occurs rapidly above this temperature (12. As in the case of other factors limiting the growth/survival of this organism. 10. lower concentrations may be effective in combination with other factors. perfringens in roast beef at 60 ºC is 14. type of solute. It grows best at pH values between 6 and 7 (the same pH as most meats). Some studies indicate that the presence of 3% NaCl delays growth of C. 10. 18). perfringens is not a tolerant organism with respect to pH. and their heat resistance is also affected by the heating substrate.8% inhibit growth of most C. perfringens in relation to food safety is the organism’s ability to grow extremely rapidly at high temperatures. At cold temperatures. etc.3.3. 10.45 ºC. (It is unusual for spores to be formed in foods after the growth of this organism (14)). 0 . although C.7 Characteristics of C.
1 Listeria spp. In foods.2. However. 229 .4.4.3 . D-values for L.4 ºC is probably a more likely minimum for growth in foods (19).19 minutes.0. respectively (21). so can be destroyed by pasteurisation.4 10. or even below 0 ºC. The upper temperature limit for the growth of L. 10. Morphology Gram-positive short rods. using hydrochloric acid as an acidulant).6 . Under ideal conditions.4 Heat resistance L. 10.3 is the lowest value where growth has been recorded.5 x 0. being potentially capable of growing . it is not a sporeformer. monocytogenes is not a particularly heat resistant organism. is between 30 and 37 ºC. and temperature. and where acetic acid is used as an acidulant. 10.4. -0.at refrigeration temperatures down to. 1.5 pH The ability of Listeria to grow at different pH values (as with other bacteria) is markedly affected by the type of acid used.2.4 . monocytogenes is reported to be 45 ºC (20). the organism is able to grow at pH values well below pH 5 (pH 4. It has been reported to have slightly greater heat resistance than certain other foodborne pathogens.0 minutes for heat resistant spores to 1. pH < 5.2 Oxygen requirements Aerobe or microaerophilic. 0. however.8 minutes for heat sensitive spores (12). 10.3 Temperature Listeria monocytogenes is unusual amongst foodborne pathogens in that it is psychrotrophic. growth at low temperatures can be very slow.98 and 0.especially at refrigeration temperatures. requiring days or weeks to reach maximum numbers.4.64. however. Its optimum growth temperature.albeit slowly . 60 and 65 ºC were reported as 10.5 .4. the lowest limit for growth is likely to be considerably higher .PATHOGEN PROFILES resistance values at 95 ºC (D-values) range from 17.23. monocytogenes in crawfish tail meat at 55. 10.0 µm.2 has been suggested as the lowest working limit (22).
Morphology Gram-negative short rods.5. 0.0 µm. at salt levels found in foods (10 . 10. 10.7 x 220.127.116.11 Salmonella spp. 10. but has been shown to be able to grow at an aw level of 0.5 .18.104.22.168 Oxygen requirements Facultative anaerobe.90. peritrichous flagella. 230 .6. 25).0 x 1 .5 (24.3 Temperature Plesiomonas can grow in the temperature range of 8 .6 10. 10.5 10.5. 10.5.1 Plesiomonas Morphology Gram-negative.45 ºC. 10.0 . or even grow. 0.83 (20).8.6 Aw/Sodium chloride L. It grows best at aw of ≥ 0.4 Heat resistance Pasteurisation at 60 ºC for 30 minutes has been reported as being effective in killing Plesiomonas shigelloides (24).3.6 Aw/Sodium chloride Plesiomonas can grow well at NaCl concentrations up to 4% (24.FISH AND SEAFOOD 10.5 pH Plesiomonas has a pH range for growth of pH 4.8 . The bacterium may survive for long periods at aw values as low as 0. 25). the optimum being 30 ºC (23). monocytogenes is quite tolerant of high NaCl/low water activities.12% NaCl or more).3 µm.5 .1.5. 10. It is likely to survive.
3 Temperature Salmonellae can grow in the temperature range of 7 . therefore. a heat resistant organism.5. the optimum being 35 . and appropriate heat treatments must be determined experimentally for low-aw foods.2 Oxygen requirements Facultative anaerobe.93 (as reported in dried meat and dehydrated soup).945. high fat or low moisture levels (low aw) will reduce the effectiveness of heat treatments. However.9.6. It is not. Salmonellae are quite resistant to drying. or possibly 0. some strains are able to grow at temperatures as low as 4 °C (26). 10.4 Heat resistance Salmonella is not a spore-forming organism. However. substrate. Salmonellae are quite resistant to freezing.5 pH Salmonella has a pH range for growth of pH 3. 10.7. and with an appropriate acid. depending on serotype.6 Aw/Sodium chloride Where all other conditions are favourable.5.37 ºC. Salmonella senftenberg 775W is about 10 . The optimal pH for Salmonella growth is between 6. although salt tolerance increases with increasing temperature (28).5 ºC.6.6. The growth of Salmonella is generally inhibited by the presence of 3 . strains vary in their ability to withstand heating.4% NaCl.0. depending on the type of acid and temperature.6. 10.6. Furthermore. D60 ºC values normally range from about 1 to 10 minutes. with a z-value of 4 .48 ºC. temperature and pH. 10. Growth is slow at temperatures below about 10 ºC.PATHOGEN PROFILES 10. 231 . pasteurisation and equivalent treatments will destroy the organism under normal circumstances. Salmonella has the potential to grow at aw levels as low as 0. Some death will occur at pH values of less than about 4. under otherwise ideal conditions.20 times more heat resistant than the average strain of Salmonella at high aw (27).8 .5 .
5 ºC. Most strains of staphylococci can grow within the pH range 4. Freezing and thawing have little effect on Staph.7.0 . aureus than citric acid).1 Staphylococcus aureus Morphology Gram-positive cocci. 10.FISH AND SEAFOOD 10.3 (optimum 7. 31).5). In meat macerate. 232 . however. with an optimum of 35 .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.2 to 9. The limits for toxin production are also narrower than for growth.7. The following relate to limits for growth only. depending on aw.3 Temperature Under otherwise ideal conditions.0. Staph. stressed cells can also be less tolerant of heating. NB: The growth of Staph. 0. 10.7 10.7 . under otherwise ideal conditions (29. aw. etc.). aureus can grow within the temperature range 7 – 48. Under most circumstances.7.4 Heat resistance Heat resistance depends very much on the food type in which the organism is being heated (conditions relating to pH. As is the case with other bacteria. the organism is heat sensitive and will be destroyed by pasteurisation. aureus viability.37 ºC (29). aureus is more limited under anaerobic than aerobic conditions. 10. but may cause some cell damage (30). aw and temperature (sensitivity to acid increases with temperature). 10. It can survive well at low temperatures.9 µm diameter. the D60 ºC value is 2 .7. fat content.7.2 Oxygen requirements Facultative anaerobe.20 minutes.7.
34).87 and > 0. pH. atmosphere.2.8 10. 31) between 0. with a temperature range for growth of 10 . An effect of increasing salt concentration is to raise the minimum pH of growth.99 aerobically under otherwise optimal conditions.4 .5) (29. especially vacuum packed foods (33) Temperature: pH: *Aw: Atmosphere: Heat Resistance: Enterotoxins are quite heat resistant.>0.6 µm 10.7.2 Oxygen requirements Facultative anaerobe. * dependent on temperature.1 Vibrio cholerae Morphology Gram-negative curved rods.5 . strain. 10.99 little or no toxin production in anaerobically packed foods.8.3 Temperature The optimum temperature for growth has been reported as 30 ºC. heating at 100 °C for at least 30 minutes may be required to destroy unpurified toxin (31.86 is the generally recognised minimum in foods (32). aureus is unusual amongst food-poisoning organisms in its ability to tolerate low water activities.7 Limits permitting toxin production 10 .8.0 (optimum 7. 233 .7. aureus can grow in 7 . 0.0. However. Staphylococci are more resistant to salt present in foods than other organisms. 10.6 Aw/Sodium chloride Staph.46 ºC (optimum between 35 and 40 ºC) (very little toxin is produced at the upper and lower extremes) (31) 5. Staph.10% NaCl.83 . but certain strains can grow in 20% NaCl.8. It can grow over the aw range 0.2 .0 . In general.8 x 1. an aw of 0. and solute.9. In general. 10.43 ºC (35).7.PATHOGEN PROFILES 10.
3 Temperature The minimum and maximum temperatures reported for growth in laboratory media.97 . declining in number during storage.4 Heat resistance V.6 Aw/Sodium chloride V.9.8. there is evidence that V. with optimum at 7. are 5 and 43 ºC.998.4.5 ºC).1 . V. cholerae is sensitive to heat at temperatures of >45 ºC. but not acid conditions.5 pH This organism is tolerant of high pH values. parahaemolyticus is easily destroyed by drying. 10. 10.8.6 (35. 10. but only moderately sensitive to freezing (40).9 10. The D-value for V.0% NaCl (35).9.9. It can grow in the presence of 0. 10. cholerae can grow over the aw range 0. 10.FISH AND SEAFOOD Also.65 minutes (35). respectively. under otherwise favourable conditions. with an optimum of 37 ºC (39). cholerae may persist on frozen fish for >180 days (36).9. with an optimum of 0.0.2 Oxygen requirements Facultative anaerobe.1 Vibrio parahaemolyticus Morphology Gram-negative curved rods.8.984 (35). and it is sensitive to refrigeration temperatures (0 . 10. 38). 234 .6. 37. cholerae in crab meat homogenate at 60 ºC is 2. it has a pH range for growth of pH 5 .
The optimum concentration of NaCl equates to aw of approximately 0.5% NaCl. its upper and lower limits for growth are 8 (although some reports suggest 10) and 0.98 (35.4 Heat resistance V.43 ºC (42).11. with an optimum pH of 7. 10. 41). and 2.02 . parahaemolyticus will grow at aw levels as low as 0.1 Morphology Gram-negative curved rods. 235 . The organism can survive for long periods at freezer temperatures (42). it has been reported that cooking oysters for 10 minutes at 50 ºC should ensure destruction of the organism (43).6 (41).8 .2 Oxygen requirements Facultative anaerobe.4 Heat resistance V. with a temperature range for growth of 8 .10. depending on solute.PATHOGEN PROFILES 10.9.10.5 pH The pH range for growth is 4.4% (39. 10.5 minutes. The D-values at 55 ºC for clam and crab homogenate are 0.3 Temperature The optimum temperature for growth has been reported at 37 ºC. parahaemolyticus are not heat resistant.0.10 Vibrio vulnificus 10.9.10. parahaemolyticus is unable to grow in the absence of salt.9. respectively (39).8. respectively.10. V. 10. 10. and are readily destroyed by cooking. vulnificus is heat sensitive. but the optimum concentration is 2 .6 . 41).922.6 Aw/Sodium chloride V. 10. 10.29 minutes.
W.5. in Foodborne Disease Handbook. 10. Stelma G. Notermans S. Marcel Dekker.. 2005.6 Aw/Sodium chloride V. New York. 1994. 3.N.. Cliver D. Juneja V. Aeromonas species. 2003.. Volume 1: Diseases Caused by Bacteria. Bier J.8 (42).. Doyle M.M.P.P. Microbiology and Molecular Biology. Potential hazards associated with REPFEDS. 279-303. 1996. Murrell K. Kim J. New York. 5.5% (42). 2. Gorman J..P.C. Marcel Dekker Inc.H. 1994. ASM Press.L. 121-76.. Faris A.. 6 (1). in Microorganisms in Foods. Stelma G... Food Control.R. Marcel Dekker.W. Lund B.0% NaCl. Marcel Dekker. 383-408. Abeyta C. Aeromonas hydrophila group. Beuchat L. Eds. Johnson E. 8. Doyle M. Eds.10. 357-67.D.. Palumbo S. Doyle M.M. Principles of control.W. Bhunia A. Novak J. in Clostridium botulinum: Ecology and Control in Foods. Smith J.R. Hauschild A. New York.. 10. in Clostridium botulinum: Ecology and Control in Foods.D.H. Austin J. Miliotis M. Eds. Gaze J.O.10.H.N.H.D. in Foodborne Bacterial Pathogens. Cliver D. Eds. Betts G. Fratamico P.. Diseases Caused by Bacteria. Washington D. 7. 1989. Eds. New York.A. Hui Y. Murrell K. 9. New York. Clostridium botulinum.R. Foegeding P.. London.E. Volume 5: Microbiological Specifications of Food Pathogens. Dodds K. New York. Growth and heat resistance of psychrotrophic Clostridium botulinum in relation to ‘sous vide’.. International Commission on Microbiological Specifications for Foods. 4. Hauschild A. Clostridium botulinum and Clostridium perfringens. Aeromonas. New York.10.FISH AND SEAFOOD 10. Montville T. 1-19. Ed.J. 5-19. References International Commission on Microbiological Specifications for Foods.5 . Marcel Dekker. Volume 1. 6..H. Clostridium botulinum. 1993. 57-63.. Gorham J. Marcel Dekker. Ed.. Great Britain. Dodds K. 97-131.. 236 . 1993. Eds.D. Ed. Blackie. Eds..H. 329-49. 2001. in Food Microbiology: Fundamentals and Frontiers. 1995. Peck M.. with an optimum pH of 7. in Foodborne Pathogens. 10. in International Handbook of Foodborne Pathogens.W.M.O. in Foodborne Disease Handbook..L. Aeromonas hydrophila.W. in Foodborne Bacterial Pathogens.5 pH The pH range for growth is 5 . Hauschild A. 11. Hui Y. Caister Academic Press. Marcel Dekker. Dodds K. 1-27.11 1. Kovacek K.K.L. 111-89. 1989. Clostridium botulinum. vulnificus grows over the range 0.. with an optimum of 2.L..
. Blackie.P. International Commission on Microbiological Specifications for Foods.P. 1991.. Ryser E.C. in Foodborne Pathogens: An Illustrated Text.. Volume 1. 141-82.An Introduction. in Foodborne Bacterial Pathogens.R. 17.A. 1989. Johnson E. International Commission on Microbiological Specifications for Foods. Doyle M. Eds. 191-243.T.J. Archer P. Ed. 1996. Washington D. Clostridium perfringens..J. Varnam A. 1996. London. in Microorganisms in Foods. Emery D. International Commission on Microbiological Specifications for Foods. Lund B.O. Diseases Caused by Bacteria. Journal of Applied Bacteriology.. 14. Evans M. 16.K.. Montville T. in Foodborne Bacterial Pathogens. Banks J. Rinke H. London. International Commission on Microbiological Specifications for Foods. Wolfe Publishing Ltd.A. 25. McClane B. Clostridium perfringens. Blackie. 133-67.H. Ed.O.A. Listeria.A. 1994. 112-25. Plesiomonas shigelloides. London. ASM Press. New York. in Foodborne Diseases. Ed. Clostridium perfringens. in Food Microbiology . Ed.PATHOGEN PROFILES 12. 208-13. 201-8. Beuchat L. 2001. 237 .J.. ASM Press. Avian Disease. 22. Hui Y. Varnam A. 137-64. Evans M. 129-38.D. Nagaraja K.A. 18.. Murrell K.M. in Foodborne Infection and Intoxication. Listeria monocytogenes. Cliver D.V.. Growth of Listeria monocytogenes at refrigeration temperatures. in Microorganisms in Foods. 2007.P.. Gorham J.. Matthews K. 1996. Marcel Dekker. Listeria monocytogenes. in Foodborne Disease Handbook. Blackie. Eds.P. New York. Plesiomonas. Ed. Walker S. 2005. Clostridium perfringens gastroenteritis. London.H. 23.. 24.. 19. 33...M. Halvorson D. Marcel Dekker. International Commission on Microbiological Specifications for Foods. Eds. 13. 15. 26. in Food Microbiology: Fundamentals and Frontiers.. Eds.R. Wrigley D. Eds. 311-25. Cliver D. Marth E. Listeriosis and Food Safety. 68 (2).J. Riemann H.G. 229-40. Kim C. Lancet.. 1990.G. Labbe R. 2006. 1989. Ed. 336 (8721). Koburger J. International Commission on Microbiological Specifications for Foods. Cliver D.G.C. 351-82. Doyle M. Foodborne disease due to Bacillus and Clostridium species. London. Washington D. Doyle M. in Microorganisms in Foods.R. Labbe R.O. Elsevier. 21. Volume 5: Microbiological Specifications of Food Pathogens. Juneja V. Clostridium perfringens. Effect of time and temperature on growth of Salmonella enteritidis in experimentally inoculated eggs. 1989.H.. London. 982-6. Plesiomonas shigelloides. 20. Clostridium perfringens food poisoning. Academic Press. 157-62. 735-42. New York. 1990. 1990. Volume 5: Microbiological Specifications of Food Pathogens. Marcel Dekker. Marcel Dekker. Volume 5: Microbiological Specifications of Food Pathogens. Montville T.H. New York.
Academic Press. Warner R. Lee-Wong A. Stewart G. Ed. Stress Response and Resistance. Ed.P. Volume 5. International Commission on Microbiological Specifications for Foods. 543-68. Crowcroft N.S. 1996. Blackie. Cliver D.. 2005. International Commission on Microbiological Specifications for Foods. 4 (13). On the resisting power of Vibrio cholerae and typhoid Bacillus against low temperatures. Bhunia A. Staphylococcus aureus.S. 217-64. Smith J. Applied and Environmental Microbiology. 13 (11). London. in Microorganisms in Foods.J. Wymondham. New York. Riemann H. Vibrio parahaemolyticus.C. Ed. Ed. 38.H. International Commission on Microbiological Specifications for Foods. London. Dairy. Cleland D. Reed G. Eds. 37.D. Doyle M.K. in Foodborne Disease. in Foodborne Infections and Intoxications.A. Salmonella. Loessner M. London.M. 8. Eds. Caister Academic Press. Twedt R.M. Wilkinson B..M. D’Aoust J-Y. Springer Science. Ed. 1930. International Commission on Microbiological Specifications for Foods. 331-57.. 327-52. Tohyama Y. 426-35. Cliver D. Distribution of Vibrio vulnificus and other lactose-fermenting vibrios in the marine environment. 1996. Doyle M. 85-106.R. Loessner M. Blackie. 31. New York. Blackie. 40. International Commission on Microbiological Specifications for Foods. in Modern Food Microbiology. Eds. 985-98. 2005. 39. Japanese Journal of Experimental Medicine. 30. 545-66. 1989. Woodhead Publishing Ltd. 642. Experimental investigations concerning the prevention of epidemic diseases due to aquatic products (Fourth Report). Salmonellae.. Foodborne illness (Part 1): Staphylococcal (“Staph”) food poisoning. 1996. Marcel Dekker.. Volume 5. 327-445.. 1990. 2005. 35. Cholera: current epidemiology. Microbiological Specifications of Food Pathogens.62.M. Oliver J. 1994. 1993. Marcel Dekker. Golden D. Golden D.P. CDR Review. 36. 1989. New York. Academic Press. 34.. London. 2005. Gustafson J. Staphylococcus aureus as a food pathogen: staphylococcal enterotoxins and stress response systems. in Understanding Pathogen Behaviour Virulence. 29. Staphylococcal Food Poisoning. in Microorganisms in Foods. Vibrio parahaemolyticus. Staphylococcal gastroenteritis. Jay J. 28. 1983. in Microorganisms in Foods.P. 33. Fratamico P. Microbiological Specifications of Food Pathogens. Food and Environmental Sanitation. Ed. Staphylococcal intoxications. 32. R15764.C.FISH AND SEAFOOD 27. Griffiths M.O. in Foodborne Pathogens. International Commission on Microbiological Specifications for Foods. 45 (3).J. Volume 5: Microbiological Specifications of Food Pathogens. Cambridge. London. 238 .A. Ed. Bergdoll M.S. in Foodborne Bacterial Pathogens. Jay J...A.O.L. 414-25. 273-84. Vibrio cholerae. 523. Bergdoll M. in Foodborne Pathogens: Microbiology and Molecular Biology..
Jay J. Cook D. Golden D. 239 .J.. 985-9.A. Golden D. 657-78. Ruple A. Vibrio vulnificus. Loessner M. 42.D. Ed. 1992. International Commission on Microbiological Specifications for Foods.. Springer Science. Blackie..M.. 55 (12). in Modern Food Microbiology. Yersinia. International Commission on Microbiological Specifications for Foods.W.J. 1996. Journal of Food Protection. Volume 5. Jay J. Foodborne gastroenteritis caused by Vibrio. Loessner M. Cold storage and mild heat treatment as processing aids to reduce the numbers of Vibrio vulnificus in raw oysters. Microbiological Specifications of Food Pathogens. London.PATHOGEN PROFILES 41. and Campylobacter species. 2005.M. in Microorganisms in Foods. 436-9.A. 43.. New York.
FISH AND SEAFOOD 240 .
org.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 Web site: www.org.gov.org/ .uk/ Department for Environment Food and Rural Affairs (Defra) Nobel House 17 Smith Square London SW1P 3JR United Kingdom Tel: + 44 (0) 207 2386000 Fax: + 44 (0) 207 2382188 241 Email: firstname.lastname@example.org@ifm.gsi.defra.gsi.gov.org.uk or v.CONTACTS Addresses of Trade Associations and Professional Bodies Association of British Salted Fish Curers and Exporters c/o Cawoods (Fish Curers) Ltd Estate Rd No 6 South Humberside Industrial Estate Grimsby North East Lincolnshire DN31 2TG United Kingdom Tel: + 44 (0) 1472 342248 Fax: + 44 (0) 1472 353100 Email: email@example.com Web site: www.scotland.ifm.uk Prepared Fish Products Association (PFPA) Federation House 6 Catherine Street London WC2B 5JJ United Kingdom Tel: + 44 (0) 208 78362460 The Scottish Government Fisheries Group Environment and Rural Affairs Department Pentland House 47 Robb's Loan Edinburgh EH14 1TY United Kingdom Tel: + 44 (0) 8457 741741 or + 44 (0)131 5568400 Fax: + 44 (0)1397 795001 Email: firstname.lastname@example.org/Topics/Fisheries Seafish Industry Authority 18 Logie Mill Logie Green Road Edinburgh EH7 4HS United Kingdom Tel: + 44 (0)131 5583331 Fax: + 44 (0)131 5581442 Web site: www.uk Web site: www.
dk Web site: www.org/fishery/ European Association of Fish Producers’ Organisations H.no/ Matís ohf (Icelandic Fisheries Laboratories) Skúlagata 4 101 Reykjavík Iceland Tel: + 354 422 5000 Fax: + 354 422 5001 Email: email@example.com Irish Sea Fisheries Board PO Box 12 Crofton Road Dun Laoghaire Co.13 Postbox 6122 N-9291 Tromsø Norway Tel: + 47 77 629000 Fax: + 47 77 629100 Email: firstname.lastname@example.org/ 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: email@example.com Web site: www.no Web site: www.difta.FISH AND SEAFOOD Department of Agriculture and Rural Development.is/ 242 .eapo.suite.org Web site: www.dk Danmarks Fiskehandlere H.asp Food and Agriculture Organization of the United Nations Fisheries and Aquaculture Department Viale delle Terme di Caracalla 00153 Rome Italy Tel: + 39 (06) 57051 Fax: + 39 (06) 57053152 Email: FAO-HQ@fao.C.firstname.lastname@example.org Web site: www.dardni.dk Web site: www.uk Web site: www. Andersens Boulevard 37.uk/index/fisheriesfarming-and-food.ie/templates/homepage.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. Dublin Eire Ireland Tel: + 353 1 2144100 Web site: www. 1 1553 København V Denmark Tel: + 45 35 372023 Fax: + 45 35 372033 Email: Info@fiskehandlerne.gov. 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: email@example.com/s Norwegian Institute of Fisheries and Aquaculture Ltd Muninbakken 9 .bim. Baelskaai 25 8400 Oostende Belgium Tel: + 32 59321876 Fax: + 32 59322840 Email: firstname.lastname@example.org.
CONTACTS Fishing Industry Research Institute 15 Lower Hope Street Rosebank 7700 Republic of South Africa National Fisheries Institute (USA) 7918 Jones Branch Drive Suite 700 McLean VA 22102 United States of America Tel: + 1 703 7528880 Email: email@example.com Web site: www.nz/sc-home Health Protection Agency (HPA) 61 Colindale Avenue London NW9 5EQ United Kingdom TeI: + 44 (0) 208 2004400 Fax: + 44 (0) 208 2007868 Web site: www.gov.ifr.org Web site: www.hpa.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.bsi-global.org.ac.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 Web site: www.gsi.au New Zealand Seafood Industry Council Private Bag 24-901 Wellington 6142 New Zealand Tel: + 64 4 3854005 Fax: + 64 4 3852727 Email: firstname.lastname@example.org Department of Health Richmond House 79 Whitehall London SW1A 2NS United Kingdom Tel: + 44 (0) 207 2104850 Email: email@example.com/en/index.cmar.htm Food and Drink Federation (FDF) 6 Catherine Street London WC2B SJJ United Kingdom TeI: + 44 (0) 207 8362460 Fax: + 44 (0) 207 8360580 Email: firstname.lastname@example.org Web site: www.org Other Sources of Information British Standards Institution 389 Chiswick High Road London W4 4AL United Kingdom Tel: + 44 (0) 208 9969001 Fax: + 44 (0) 208 9967001 Email: email@example.com Web site: www.org.csiro.co.seafood.com/ 243 .nz Web site: www.gov.fdf.
org Web site: www.int.org.uk/ Health Protection Scotland Clifton House Clifton Place Glasgow G3 7LN United Kingdom Tel: + 44 (0)141 3001100 Fax: + 44 (0)141 3001170 Email: firstname.lastname@example.org Web site: www.codexalimentarius.int/foodsafety World Health Organization (WHO) Headquarters CH .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/scieh.who.se/ FAO/WHO Collaborating Centre for Research and Training in Food Hygiene and Zoonoses Robert von Ostertag Institute Postfach 330013 D-14191 Berlin Germany Tel: + 49 (0) 188 84122155/ 2157 Fax: + 49 (0) 188 84122957 Email: firstname.lastname@example.org Web site: http://www. jsp Food Safety 10 .1211 Geneva 27 Switzerland Tel: + 41 22 7912111 Fax: + 41 22 7913111 Email: email@example.com/ SIK .se Web site: www.net/web/indexen.hps.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.00187 Rome Italy Tel: + 39 06 487751 Fax: + 39 06 4877599 Email: firstname.lastname@example.org 244 .nhs.co.euro.php Society of Food Hygiene and Technology The Granary Middleton House Farm Tamworth Road Middleton Staffs B78 2BD United Kingdom Tel: + 44 (0) 1827 872500 Fax: + 44 (0) 1827 875800 Email: email@example.com/index.nhs.who. Web site: www.scot.co.euro.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 Web site: www.asp Codex Alimentarius Commission (CAC) Viale delle Terme di Caracalla 00153 Rome Italy Tel: + 39 (06) 57051 Fax: + 39 (06) 57054593 Email: Codex@fao.sfam.who.uk Web site: www.
htm Centre for Disease Control and Prevention (CDC) (US) http://www.ie/ Food Science Australia http://www.ift.sofht.int/foodsafety/en/ European Commission: Activities of the European Union food safety http://europa.org Web site: http://www.co.org/ Royal Institute of Public Health (UK) http://www.Center for Food Safety and Applied Nutrition 5100 Paint Branch Parkway College Park MD 20740-3835 United States of America Tel: + 1 888 SAFEFOOD (+ 1 888 723 3366) Web site: www.riph.foodprotection.foodscience.org/ Institute of Food Technologists http://www.org.cfsan.CONTACTS US FDA .who.gov/ Institute of Food Technologists 525 W.fpa-food.cdc.uk/ Society of Food Hygiene and Technology (UK) http://www.fsai.gov/ World Health Organization (WHO): Food safety programmes and projects http://www.eu/pol/food/index_en.org/ The Association of Food.gov/foodsafety/ Food Safety Authority of Ireland http://www. Van Buren Suite 1000 Chicago IL 60607 United States of America Tel: + 1 3127828424 or + 1 8004383663 Fax: + 1 3127828348 Email: email@example.com/ 245 . Beverage and Consumer Products Companies http://www.au/ International Association for Food Protection http://www.ift.FoodSafety.org/cms/ Useful Web Sites Gateway to Government food safety information (US) http://www.csiro.
FISH AND SEAFOOD 246 .
as TTX producers 145 Amnesic shellfish poisoning (ASP) toxins 146-8 caused by domoic acid in crustacean shellfish 87 Anchovies. in crustacean shellfish 87 Alteromonas spp.INDEX Achromobacter spp. as spoilage organisms in chilled recipe meals 43 growth and survival in chilled and frozen raw fish 17 pathogen profile 225-6 pathogenic microorganisms in molluscan shellfish 59-60. preserved by dry/wet salting 99 Anisakis spp. breakdown of – as indicator of fish freshness 10-1 Azaspiracids 158-9 detection methods 160 origins and distribution 159 toxicology 159-60 Bacalao 97. as autochthonous bacterial flora of fish 3 Aeromonas spp. source of PSP toxins 142 Alexandrium ostenfeldii. growth and survival in crustacean shellfish 86 Air drying 101 Alcaligenes. 69-70 Aeromonas hydrophila. 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. as spoilage organism in oysters 65 as spoilage organism in tropical crustacean shellfish 81 in molluscan shellfish 60 Alexandrium. 102 Bacillus spp. causing spoilage of dried fish 110. pathogenic microorganisms in molluscan shellfish 59-60 as contaminant in enrobing process 37 as risk in breaded shrimp and prawn products 82 growth and survival in chilled and frozen prepared fish products 45 growth in vegetable and rice 247 . growth and survival in chilled and frozen raw fish 19 of concern in preserved fish 115 Aspergillus spp. 111 Astroviruses. to test microbiological criteria 19 Aeromonadaceae. growth and survival in crustacean shellfish 86 Aeromonas sobria. in crustacean shellfish 86-7 ATP. planktonic source of spirolides in shellfish 163 Allergens. as autochthonous bacterial flora of fish 3 in fish sauce 127 Bacillus cereus.
preserved by marinating 100 Chemical detection methods. importance in lactic acid fermentation in fish products 130. of fish – effects on microflora 5-6 Carbohydrate. 98-9 Brochothrix thermosphacta.FISH AND SEAFOOD components of seafood ready meals 40 Bacterial pathogens. 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 . 113. as gas in modifiedatmosphere packaging 6 CCP Decision Tree (in HACCP plan) (Fig) 185 Ceviche. 68-9 Capture and handling. cause of neurotoxin shellfish poisoning 153-5 origins and distribution 1545 toxicology 155 importance in wet salting of fish 96 Brining 93. concern with respect to use of MAP 7 growth and survival in chilled and frozen prepared fish products 45 growth and survival in chilled and frozen raw fish 15-6 growth and survival in crustacean shellfish 84 growth and survival in cured. affected by azaspiracids 159 affected by cyclic imines 161 Botulism. of recipe fish meals 43 Cholera. in crustacean shellfish 86-7 Campylobacter coli. 137. mould causing spoilage of dried fish 110 Battering process 36-8 Bivalves. as cause of spoilage of MAP fish 15 Calciviruses. 136. associated with consumption of fish/fish products 16. description and uses 56-7 effects of processing on microflora 60-4 hot-smoking 107 Clear smoke treatments 108 Clostridium. in crustacean shellfish 87 Chilled prepared fish products 27-51 Chilled raw fish 1-25 Chilled storage. as autochthonous bacterial flora of fish 3 Clostridium botulinum. of fresh fish spoilage 11 Chemical pollutants. growth and survival in crustacean shellfish 86 growth and survival in molluscan shellfish 59-60. for PSP toxins 144 Chemical indicators. fish sauce 123 Basipetospora halophila. growth and survival in crustacean shellfish 86 pathogenic microorganisms in molluscan shellfish 59-60 Campylobacter jejuni. caused by Vibrio cholerae O1 17 Ciguatoxins 155-8 detection methods 157-8 origins and distribution 1567 toxicology 157 Clams. 131 Carbon dioxide. 226 Breading process 36-8 Brevetoxins. indigenous – in fish (Table) 4 non-indigenous – in fish (Table) 4 Bakasang.
cooked – effects of processing on microflora 83 forms for purchase 80 freshness indicators 83 Crayfish. to determine fresh fish spoilage 11 Corynebacteria. 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. of prepared fish products 44 Escherichia coli. to detect NSP toxins 155 ELISA kit (Biosense). growth and survival in crabmeat 84 growth and survival in cured. hot-smoking 107 ELISA assay. for detection of DA 148 ELISA kit (DSP-Check). as autochthonous bacterial flora of fish 3 Crabs. 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. smoked and dried fish 114 pathogenic microorganisms in molluscan shellfish 59-60 249 . growth and survival in chilled and frozen raw fish 19 of concern in cold-smoked salmon 115 Domoic acid (DA). processing 334 Cockles. 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. of fish – analysis of 10 Diarrhetic shellfish poisoning (DSP) toxins 148-51 detection methods 150-1 origins and distribution 14950 toxicology 150 Dinophysis spp. description and uses 59 effects of processing on microflora 60-4 Cold-smoking 104-5. smoked and dried fish 114 Enzymic spoilage. 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. 152 Diphyllobothrium. 149-50. source of azaspiracids 159 source of DSP toxins 148-9. to detect DSP toxins 150-1 Enrobing process 36-8 Enterobacteriaceae. 106 Contacts 241-5 Conway method. adverse condition on poorly preserved fish 110 EC grading scheme. to determine fresh fish spoilage 11 Eels. as spoilage organisms in tropical crustacean shellfish 81 Coryneforms. as contaminants in scampi 36.INDEX pathogen profile 226-7 Clostridium perfringens. of fish – different processes 94 Dun.
of prepared fish products 44 Gambiertoxins.FISH AND SEAFOOD EU legislation. definition of terms 176-7 implementation and review of plan 184-6 in fish and seafood product manufacture 175-88 seven stages of system 17784 Halobacterium spp. caused by Photobacterium phosphoreum 15 Hon kare bushi 107 Hot-smoking 105-7 250 . fermented fish product 124 General Pathogen Surveillance Programme 84 Gravad fish. spoilage 9-11 Freshness grading schemes 9 Frozen prepared fish products 27-51 Frozen raw fish 1-25 Frozen storage. to measure fish skin resistance 10 Flavobacterium. of chilled and frozen prepared fish products – preparation 33-6 Fish sauces 123-4 Fish sauces and pastes. source of PSP toxins 142 Gyoshoyu. causing pink condition on fish surface 110 in fish sauce 127 Halococcus spp. definition 1 Fermented fish 123-40 Fermented fish/salt products of Thailand (Table) 125 Fish and shellfish toxins 141-74 Fish cakes. 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. preserved by marinating 101 Gravlaks. in crustacean shellfish 86-7 in molluscan shellfish 59-60 Herring. specific EU hygiene rules 196-224 Fresh fish. member of cyclic imine group 161 Gymnodinium catenatum. fish sauce 123 HACCP. of fresh fish spoilage 911 Evisceration. for evaluation of fish quality 9 Eurotium. processing 35 Fish component. of fish at sea 5 Fatty fish. 102 Gymnodimines. as source of inulin in lactic acid fermentation in fish products 134 Garos. concerning crustacean shellfish 87-8 European Grading Scheme. causing pink condition on fish surface 110 in fish sauce 127 Handling. as source of ciguatoxins 156 Garlic. preserved by marinating 100 preserved by dry/wet salting 99 Histamine fish poisoning. mould causing spoilage of dried fish 110 Evaluation. 130-5 Fish-spoilage bacteria 12 Fishtester. of fish – effects on microflora 5-6 Hepatitis (non-A. fermented fish product 126 Green cured split cod 97. effects of processing 127-30 Fish sauces and pastes of south-east Asia (Table) 124 Fish/salt/carbohydrate products of Asia (Table) 125. in crustacean shellfish 86-7 in molluscan shellfish 59-60 Hepatitis A virus. non-B) viruses.
smoked and dried fish (Table) 116-7 for shellfish products 70 Microbiological limits. as hazard in regard to reef-caught fish 28 growth and survival in crustacean shellfish 84 pathogen profile 229-30 Listeria monocytogenes. 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. initial – of chilled and frozen prepared fish products 31-2 of chilled and frozen raw fish 3-5 251 . concerning crustacean shellfis 88 Jellied eels. leading to oxidative rancidity 11 Lipophilic shellfish poisoning toxins 149.INDEX Hygiene regulations for foodstuffs production and distribution (EU) 190-6 Ice storage. cooked – effects of processing on microflora 83 forms for purchase 80 freshness indicators 83 Mackerel. smoked and dried fish 114-5 hazard in cold-smoking 105 hazard in mild-processed marinated products 101 Listeriosis. as autochthonous bacterial flora of fish 3 Lean fish. hot-smoking 107 preserved by dry/wet salting 99 Marinating 94. 151-3 detection methods 153 Liquid smoke 107-8 Listeria spp. in fish fillet 1 Lipid oxidation. producer of brevetoxins 154 Katsuobushi 107 Kench curing 97-8 Klipfish 97 Lactic acid bacteria. to reduce rate of fish spoilage 12 Ikan kaju. 135 I-zushi. causing spoilage in vacuum-packed dried salmon 111 Lactic acid fermentation. for chilled and frozen prepared fish products 47-9 for chilled and frozen raw fish 19-20 for crustacean shellfish 87-8 for cured. 100-1 Masmeen 107 Matjes. outbreak caused by gravid 115 Lobster. for fresh and frozen fish (Table) 20 Micrococcus spp. preserved by marinating 101 Microbiological criteria. food hygiene 189-224 Lingulodinium polyedrum. preserved by salt-boiling 100 Ionising radiation. for microbial inactivation in fish products 127. growth and survival in chilled and frozen prepared fish products 46 growth and survival in cured. enabled by carbohydrate source in fish products 130. as source of yessotoxins 152 Lipid content. definition 1 Legislation. 131 Lactic fermented fish products (Table) 126 fish species used in the Philippines (Table) 126 Lactobacillus. preserved by salt-boiling 100 Karenia brevis. as source of botulism 136 International legislation.
as TTX producer 145 Norovirus. of crustacean shellfish – effects of processing 81-3 of cured. shigelloides. 153 for detection of LST toxins 151 for detection of NSP toxins 155 for detection of pinnatoxins and pteriatoxins 165 for detection of PSP toxins 144 for detection of spirolides 164 for detection of tetrotoxin 145 Mussels. 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. caused by lipid oxidation 11 Oysters. contamination leading to illness 68 description and uses 57 effects of processing on microflora 60-4 hot-smoking 107 spoilage organisms 65 P. reduction in smoking process 108 Norcardiopsis dassonvillei. smoked and dried fish 95 of fermented fish 126-7 of molluscan shellfish 59-60 Microflora. in crustacean shellfish 867 Norwalk virus. for detection of azaspiracids 160 for detection of ciguatoxins 157 for detection of DA 148 for detection of DSP toxins 150. pathogenic microorganisms in molluscan shellfish 59-60 Opisthorchis viverrini. growth and survival in chilled and frozen raw fish 17 Paralytic shellfish poisoning (PSP) toxins 142-4 detection methods 144 toxicology 143 252 . smoked and dried fish – 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 AND SEAFOOD of crustacean shellfish 80-1 of cured. contaminated by azaspiracids 158 description and uses 58 effects of processing on microflora 60-4 hot-smoking 107 Nam-budu. as autochthonous bacterial flora of fish 3 as spoilage organism in temperate crustacean shellfish 80 in molluscan shellfish 60 Mouse bioassay.
growth and survival in chilled and frozen raw fish 19 growth and survival in crustacean shellfish 86 growth and survival in cured. mould causing spoilage of dried fish 110 Potato. as source of DSP toxins 148-9 Protein content. in fish fillet 1 Proteus. 197 Plesiomonas. detection methods 165 origins and distribution 165 toxicology 165 Pissala. planktonic source of azaspiracids 159 Pseudomonadaceae.INDEX Parasites. as source of yessotoxins 152 Protoperidinium cassipes. fermented fish product 124 Plankton. as component for seafood ready meals 40 Prawns. as component for seafood ready meals 40 Pathogen profiles. reduction in smoking process 103. 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. preserved by salt-boiling 100 Pink. 163. smoked and dried fish 115 Pasta. pathogen profile 230 in molluscan shellfish 5960. spoilage organism in oysters 65 Protoceratium reticulatum. 100-1 Pindang. smoked and dried fish 113-6 growth and survival in fermented fish 136-7 growth and survival in molluscan shellfish 66-70 Pectenotoxins 151. effects of processing on microflora 81-2 freshness indicators 83 Pre-column derivatisation. in molluscan shellfish 60 Pseudomonas spp. as autochthonous bacterial flora of fish 3 as spoilage organism in chilled products 43 as spoilage organism in crustacean shellfish 80. 108 Polypaecilum pisce. 225-39 Pathogens. as cause of histamine fish poisoning 15 as spoilage organism in chilled products 43 as spoilage organism in fresh fish 14-5 Pickling 94. causing spoilage of dried fish 110-1 Photobacterium phosphoreum. 145-6 definitions 28-30 non-seafood ingredients 32 Processing. 152-3 Penicillium spp. 81 as spoilage organism in fresh fish 13 as spoilage organism in oysters 65 as TTX producers 145 253 . adverse condition on poorly preserved fish 110 Pinnatoxins member of cyclic imine group 164. contact details 241-2 Prorocentrum spp. causing spoilage in vacuum-packed dried fish 111 Pseudomonads. 161. for detection of PSP toxins 144. 69-70 Polycyclic aromatic hydrocarbons. as source of toxins 159. effects on microflora of fish 5-9 Professional bodies.
FISH AND SEAFOOD inhibition of growth by carbon dioxide 6 Pseudonitzschia multiseries. as source of tetrotoxin 144-5 Putty fish. as hazard in regard to reef-caught fish 28 growth and survival in crustacean shellfish 84 growth and survival in cured. smoked and dried fish 114 pathogen profile 230-1 pathogenic microorganisms in molluscan shellfish 59-60 Salt burn. for evaluation of fish quality 9 Radiation. processing and effects on microflora 60-1 Shewanella. processing 34. fermented fish product 126 Raphidophytes. initial microflora 31 Seafood ready meals 39-40 Seafood recipe meal centres 38-9 Shaped fish products. flow diagram (Fig. detected in cod 115 Pteriatoxins. 36 Seafood.) 41 Recipe fish products 38-40 Rhodotorula rubra. in molluscan shellfish 60 Rice. producers of brevetoxins 154 Raw fish. for microbial inactivation in fish products 127. preserved in saturated brine 99 Sauce. 100 Salting 97-9 definitions 93-4 Sardines. as spoilage organism in chilled products 43 as spoilage organism in fresh fish 13-4 as spoilage organism in temperate crustacean shellfish 80. resulting from poorly salted cod 110 Pyrodinium. to measure fish skin resistance 10 Salmon. for fish ready meals – manufacture 40-2 Saxitoxin (STX) 142-3 Scallops. implicated in amnesic shellfish poisoning 146-7 Pseudoterranova. chilled and frozen 1-25 Recipe dish. as carbohydrate source in lactic acid fermentation in fish products 134 as component for seafood ready meals 40 RIDASCREEN. as autochthonous bacterial flora of fish 3 inhibition of growth by carbon dioxide 6 Shewanella putrefaciens. 135 Rakefisk. vacuum packing 105 Salmonella spp. source of PSP toxins 142 Quality Index Method. processing 35 Shellstock. 83 254 . importance in lactic acid fermentation in fish products 130 importance in salting. assembly and packaging. hazard in salting of fish 97 Salt concentration. 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. description and uses 58-9 Scampi. flow diagram (Fig. growth and survival in chilled and frozen raw fish 19 Pseudoterranova dicipiens. fermented fish product 126 as source of botulism 136 Rakørret.) 41 ingredients preparation. smoking and marinating of fish 96 Salt-boiling 94.
smoked and dried fish 114 pathogen profile 232-3 pathogenic microorganisms in molluscan shellfish 59-60 potential for growth in pasta component of seafood ready meals 40 Stockfish. of dried fish 111 of fish 6-7 of fish – (US) NACMCF recommendations 7-8 of fish – Codex Committee 255 . as 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. of crustacean shellfish 80. detection methods 164 member of cyclic imine group 162-4 origins and distribution 163 toxicology 163-5 Spoilage. contact details 241-2 Trematode infections foodborne 137 Trichosporon. as measure of fish degradation 10 Sliming. in poorly salted cod 110 Smoked fish 93-121 Smoking 103-8 different processes 94-5 Snow Mountain virus. grading of 31 hot-smoking 107 US legislation. fish sauce 123 Shrimps. pathogenic microorganisms in molluscan shellfish 59-60 Specific spoilage organisms (SSOs) 12-5 Spirolides. fermented fish product 126 Taste panel evaluation. to measure fish skin resistance 10 Total Volatile Base-Nitrogen. as indicator of fresh fish spoilage 11 Toxins. fish and shellfish 141-74 Trade associations. 14 Torry Scale. definitions of forms for purchase 79-80 effects of processing on microflora 81-2 freshness indicators 83 Shucked shellfish. processing and effects on microflora 63-4 Skin resistance. smoked and dried fish 114 pathogenic microorganisms in molluscan shellfish 59-60 Shottsuru. of fish after capture 5 Surstrømming. 13.INDEX Shigella spp. concerning crustacean shellfish 89 Vacuum packaging. in molluscan shellfish 60 Tuna. 12-5 of recipe fish meals 43-4 of shellfish 65 Staphylococcus. role in fish degradation 10. 101 Storage. of cooked fish 9-10 Tetrotoxin 144-6 detection methods 145-6 toxicology 145 TMAO. 83 of fermented fish 135-6 of fish subjected to different preservation methods 108-10 of fresh fish 9-11. growth and survival in chilled and frozen prepared fish products 46 growth and survival in cured.
FISH AND SEAFOOD guidance 7 of fresh chilled products 42 Vegetables. 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 molluscan shellfish 67 pathogen profile 235-6 Vibrionaceae. growth and survival in chilled and frozen prepared fish products 47 growth and survival in crustacean shellfish 86-7 Wallemia sebi. causing dun condition on dried fish 110 Wet salting 98-9 Wet storage. growth and survival in molluscan shellfish 67-8 Vibrio parahaemolyticus. of shellstock 61. growth and survival in crustacean shellfish 85 pathogen profile 233-4 Vibrio cholerae 01. processing for seafood recipe meal centres 39 Vibrio spp. 68 Vibrio cholerae non-01. growth and survival in chilled and frozen raw fish 17 Vibrio cholerae 01 and 0139. 81 as TTX producers 145 pathogenic microorganisms in molluscan shellfish 59-60 risk in cold-smoking 105 Vibrio cholerae. as hazard in regard to reef-caught fish 28 as spoilage organism in crustacean shellfish 80. growth and survival in chilled and frozen raw fish 16-7 growth and survival in crustacean shellfish 86 growth and survival in fermented fish 136 growth and survival in molluscan shellfish 66-67 pathogen profile 234-5 Vibrio vulnificus. as spoilage organisms in chilled recipe meals 43 Yessotoxins 151. 62 Yersinia spp. as autochthonous bacterial flora of fish 3 as spoilage organisms in chilled products 43 as spoilage organisms in vacuum-packed dried fish 111 Viruses.