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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
and growth characteristics for key pathogens associated with the product. for his scientific input and Victoria Emerton. both in the UK and elsewhere. Alison Turner. thank you for all the support. and Meat Products. which has been the chief factor in ensuring the completion of this book. Technical Team Leader for her valuable editorial contribution. Training Manager. My gratitude is extended to Dr Peter Wareing. published by Leatherhead Food International and RSC Publishing.FOREWORD The Microbiology Handbook series includes Dairy Products. and a detailed list of contacts for various food authorities. guides to HACCP. potential hazards associated with the consumption of a range of products. The books in the series are designed as easy-to-use guides to the microorganisms found in foods. and Ann Pernet for the work put into typesetting and indexing. All handbooks also contain a review of the related legislation in Europe and UK. Each book provides a brief overview of the processing factors that determine the nature and extent of microbial growth and survival in the product. Rhea Fernandes Leatherhead Food International v . Acknowledgements All the authors involved in writing the first edition of this book are gratefully acknowledged. Fish and Seafood. I thank the contributing authors for their collaboration. I also acknowledge and thank Jackie Apps.Goldwyn. I dedicate this book to my husband . The books are intended as a source of information for microbiologists and food scientists working in the food industry and responsible for food safety.
2 Definitions 2.8 References 1.1 Definitions 4.2 Initial Microflora 4.5 Pathogens: Growth and Survival 3.7 References 4.3 Processing and its Effects on the Microflora 1.4 Spoilage 4.4 Spoilage 1. vii .6 Published Microbiological Criteria 3.1 Introduction 2.5 Pathogens: Growth and Survival 4. 4.7 Published Microbiological Criteria 1.8 References MOLLUSCAN SHELLFISH 3.3 Processing and its Effects on the Microflora 3.4 Spoilage 3.6 Pathogens: Growth and Survival 2.3 Initial Microflora 2.1 Definitions 1.7 Published Microbiological Criteria 2.7 References 3.5 Spoilage 2. 3.5 Factors Affecting Fresh Fish Spoilage 1.2 Initial Microflora 3.8 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 2.4 Processing and its Effects on the Microflora 2.9 Further Reading CHILLED AND FROZEN PREPARED FISH PRODUCTS 2.6 Pathogens: Growth and Survival 1.8 Further Reading CRUSTACEAN SHELLFISH 4.6 Published Microbiological Criteria 4.CONTENTS CONTRIBUTORS FOREWORD INTRODUCTION 1.2 Initial Microflora 1.3 Processing and its Effects on the Microflora 4. CHILLED AND FROZEN RAW FISH 1.1 Definitions 3.
7 References 5. CURED.3 Tetrodotoxin 7. 9. 14 (Hygiene requirements specific to the UK) 221 9.6 Regulation (EC) No.5 References 93 93 95 96 108 113 116 117 120 123 123 126 127 135 136 137 140 141 141 142 144 146 148 151 153 155 158 160 165 166 175 175 176 177 184 186 6.4 Amnesic Shellfish Poisoning (ASP) Toxins 7.4 Spoilage 5.5 Pathogens: Growth and Survival 6.1 Definitions 6.3 Regulation (EC) No.2 Initial Microflora 6. 8.4 Spoilage 6.3 Stages of a HACCP Study 8.6 Published Microbiological Criteria 5.3 Processing and its Effects on the Microflora 6.8 Guidance 222 9. SMOKED AND DRIED FISH 5.2 Paralytic Shellfish Poisoning (PSP) Toxins 7.8 Further Reading FERMENTED FISH 6.5.7 Neurotoxin Shellfish Poisoning (NSP) Toxins 7.1 Definitions 5.3 Processing and its Effects on the Microflora 5.9 Azaspiracids 7.12 References HACCP IN FISH AND SEAFOOD PRODUCT MANUFACTURE 8. EU FOOD HYGIENE LEGISLATION 189 9.7 Food Hygiene (England) Regulations 2006. 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 9.2 Legislative Structure 190 9.8 Ciguatoxins 7.5 Diarrhetic Shellfish Poisoning (DSP) Toxins 7.4 Regulation (EC) No.9 Other Relevant Legislation 223 9.10 Further Reading 223 viii .1 Introduction 8. S. 2073/2005 on Microbiological Criteria for Foodstuffs 214 9.2 Definitions 8. 853/2004 Laying Down Specific Hygiene Rules for Food of Animal Origin 196 9.11 Conclusion 7.6 References 6.5 Pathogens: Growth and Survival 5. 2006 No.10 Cyclic Imines 7.6 Lipophilic Shellfish Toxins (LST) 7. 7.1 Introduction 189 9.2 Initial Microflora 5.7 Further Reading FISH AND SHELLFISH TOXINS 7.1 Introduction 7.I. 852/2004 on the General Hygiene of Foodstuffs 191 9.4 Implementation and Review of the HACCP Plan 8.5 Regulation (EC) No.
5 Plesiomonas 10.8 Vibrio cholerae 10.4 Listeria spp.10 Vibrio vulnificus 10.9 Vibrio parahaemolyticus 10.7 Staphylococcus aureus 10.6 Salmonella spp. 10.11 References 225 225 226 227 229 230 230 232 233 234 235 236 241 247 CONTACTS INDEX ix . 10. 10.10 PATHOGEN PROFILES 10.2 Clostridium botulinum 10.3 Clostridium perfringens 10.1 Aeromonas spp.
in Handbook of Food Products Manufacturing: Health. The Microbiology Handbook. Safety and Processing of Wild and Farmed Fish. Luten J.Fish and Seafood consists of the microbiology of seven different product categories: chilled and frozen raw fish. Chandan R... xi . Hoboken. fermented fish. Eds. Often contamination is from human and animal sources. Wiley. Jacobsen C.. Key changes in this edition are the recent regulatory changes pertaining to food hygiene and microbiological criteria for foodstuffs.M.INTRODUCTION Fish and seafood are a main source of animal protein in the diet. Clark S. thus. Wageningen Academic Publishers. and fish and shellfish toxins.. and.B. chilled and frozen prepared fish. Catches can be gathered from seas. Geographical region. Poultry. molluscan shellfish.A. for fish. Because of their health advantages over red meats. The second edition of this handbook is a review of the entire book for currency of information. Nollet L. the consumption of fish and seafood has increased. Cross N. season. Meat. Further Reading General Fish/Seafood Microbiology Cross N. 2006.. Seafood and Vegetables. crustacean shellfish. Hui Y.. Oehlenschlager J. fish and seafood can be involved in the transmission of pathogenic microorganisms and toxins. 94767. Bekaert K. Seafood Research from Fish to Dish: Quality. Wageningen.H.. smoked and dried fish. 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. cured. Seafood processing: basic sanitation practices. Milk. rivers and lakes whose water can range from pristine to contaminated. Saebo A.. 2007.
London. 318-23.Y. Microbiological spoilage of fish and fish products. Huss H. Noordwijkerhout. Amsterdam.H. 2004.R. Hui Y. Ed. 60 (4). Modified-atmosphere packaging of fish and fish products. Ed. Seafood processing: basic sanitation practices. Smith J. 2002. 1998. 1997. Ed. Johnson J. Stanfield P.A. Stanfield P. Eds. Luten J. 431-43. Huss H. 1996. Vo1ume 39. Heinitz M. Nip W-K. November 1995. 61 (3). Gram L. Dalgaard P..L.A. Safety and Quality Issues in Fish Processing. 8 (2). Blackwell Publishing. 1997. in Kosher Food Production.H. xii . and Clostridium botulinum in smoked fish and shellfish. Huss H. Fish and fish products (Microorganisms in fish products).. Davies A. Proceedings of the International Seafood Conference... in Food Plant Sanitation. Proceedings of the International Seafood Conference.. in Seafood from Producer to Consumer: Integrated Approach to Quality. 167-77. in Microorganisms in Foods 6: Microbial Ecology of Food Commodities.L. Dalgaard P.. 121-37. London.Y..S. Marcel Dekker. Hall G.. Journal of Food Protection..S. McCarthy S. 174-249.. Luten J. 2005. Seafood: processing. Bengtsson N. Biogenic amines in fish and shellfish. Salmonella spp. Eds. 245-66 Bremner H. 1996. New York. Woodhead Publishing Ltd.E. in Fish Processing Technology. Incidence and survival of Listeria monocytogenes in ready-to-eat seafood products. Blech Z.P. Spoilage and shelf-life extension of fresh fish and shellfish. Kluwer Academic / Plenum Publishers. Control of indigenous pathogenic bacteria in seafood. 329-65.N. International Commission on Microbiological Specifications for Foods. Oxford. Cambridge.. 459-72.M. Oxford. in Food Processing: Principles and Applications. Tong P. 87121.. 413-30.M. Gram L. Simpson B.H. Woodhead Publishing Ltd. Taylor S.B. Microbiology of fish and fish products.. 2002. Hui Y. Predictive microbiological modelling and seafood quality.. 543-61. Gudmundsson M. Minimal processing in practice: seafood. 372-6.J. Ashie I. basic sanitation practices. Amsterdam.International Commission on Microbiological Specifications for Foods. Cambridge. Blackie.K.H.L. Martin R..B. 1997. Bruinsma B.A. Blech Z. 1997. Ed. 1997. Noordwijkerhout. Blackwell Publishing. Flick G. Rawles D. The incidence of Listeria spp. Eds. Elsevier. Academic Press. in Seafood from Producer to Consumer: Integrated Approach to Quality. 2003. CRC Critical Reviews in Food Science and Nutrition.. 33 (1). 36 (1-2).R. Gorham J. Journal of Food Protection.. Elsevier.H. Ed.D. 2004. Ohlsson T.. in Advances in Food and Nutrition Research. November 1995. International Journal of Food Microbiology. Food Control. New York. Ed. The fish industry. Smith J. 200-23. Ventresca P. 1996. in Minimal Processing Technologies in the Food Industry. Hafsteinsson H. 91-8.
Huss H. Fish spoilage and safety. Copenhagen. Huss H. Blackie. Eds. 1991. 1994. 11 (2). Institut fur Veterinarmedizin.H. 90-106. Twigg B. August 1991. Liston J. Eds. Gibson D. 16 (1). 1990. Elsevier. Dalgaard P.H. Copenhagen. Food Science and Technology Today.D. 118-20.M. 1992. in Food and Beverage Technology International: USA. Fish: microbiological spoilage and safety. 319-33.R. Jakobsen M. Ed.M. 1992.. Microbial hazards of seafood consumption.. Gram L. in Quality Attributes and Their Measurement in Meat. 44 (12). in Fisheries Processing: Biotechnological Applications. Elsevier. xiii . Bonnell A. Bacterial spoilage of seafood. 1996. Garrett E. Activities Report of the R&D Associates. 1995. 26 (3). Ed. Ed..H. London. 5662. McMeekin T. August 1991.R. 46 (1). Poultry and Fish Products. International Commission on Microbiological Specifications for Foods.J. Martin A.. 459-78. Liston J. 166-73. 1-17. Berlin. Huss H. International Journal of Food Microbiology. 25-39. Gilbert R. Jakobsen M. poultry and fish. Eds. 1995. Hobbs G. Glasgow. Chapman and Hall. in Public Health Aspects of Seafood-borne Zoonotic Diseases. in Quality Assurance in the Fish Industry: Proceedings of an International Conference. in Microorganisms in Foods.M.. 1992. Ed. Evaluation of the bacteriological quality of seafood. 265-79. guidelines. Jakobsen M.A. Microbiological indicators and seafood processing. 359-403. August 1991. Microbial growth and its control in meat. London. Amsterdam. 2257. Ward D. 93-105. Food Technology. and specifications for seafood products.. Modelling of microbial activity and prediction of shelf-life for packed fish. A new look at microbiological standards. Sofos J. 54-61.. 305-17. Eds. Ross T. Copenhagen. Pearson A. Bonnell A. 1994.S. Amsterdam. Application of predictive microbiology to assure the quality and safety of fish and fish products. Microbiological quality of fishery products. Dutson T. 121-8. The microbiology of seafood products. Seafood toxins of microbiological origin. 26 (3). Ward D.R. Olley J. Bernoth E-M. 1994.International Commission on Microbiological Specifications for Foods. Elsevier. International Journal of Food Microbiology. in Quality Assurance in the Fish Industry: Proceedings of an International Conference. 1992. Sterling Publications Ltd. 1990.. Qualitative and quantitative characterization of spoilage bacteria from packed fish. 5 (3). Gram L. London. Evaluation of the bacteriological quality of seafood. Ed. in Quality Assurance in the Fish Industry: Proceedings of an International Conference. Bacterial pathogens transmitted by seafood. PHLS Microbiology Digest. London. Liston J. Dalgaard P. 1994.N. 1994. Amsterdam. Liston J. International Journal of Food Microbiology. Blackie. 269-82.. Liston J. 1989. Volume 5: Microbiological Specifications of Food Pathogens. Chapman & Hall.A. in Quality Assurance in Seafood Processing: A Practical Guide.D.
Hodgkiss W. in Microbiology of Frozen Foods. Hackney C. London. 42 (3). Regenstein J.Pedrosa-Menabrito A.. 8-16. Journal of Food Quality. Dicharry A. 1987. 71-117. Microbiology of fish. Volume 1.. Hobbs G.K. Part I . Elsevier Applied Science Publishers. London. Wiley. Applied Science Publishers. Food Technology. 46 (1). Robinson R. 42 (3). 1985.K. Seafood-borne bacterial pathogens of marine origin. Microbiology of frozen fish and related products. 117-27. Shelf-life extension of fresh fish . Microbiological hazards associated with fishery products. Microbiological standards.. in Essays in Agricultural and Food Microbiology. Food Technology. guidelines. and specifications and inspection of seafood products.R. Hobbs G. Ed. xiv .C. 169-208. 103.spoilage of fish. Ed. Ed.. Eyles M. 1986. 1988. Davies R.a review. in Developments in Food Microbiology. 104-9.S. 199-226. 1988. The bacteriology of fish handling and processing. CSIRO Food Research Quarterly. 11 (2). Chichester.J. Lamprecht E.R. Norris J. Garrett E. Simmonds C. 90-3.M. 1988. 1982.
This super class of vertebrates includes all the bony and cartilaginous finfish. and other forms of aquatic animal as part of their ‘fish’ definition. respectively (1). Fish fillet composition can vary significantly within the same species due to feed intake. offering less than 0. migratory patterns. Protein content in fish fillet varies typically from 16 . Fish are an important part of a healthy diet since they contain high quality protein.1 Definitions Fish are classified as any of the cold-blooded aquatic vertebrates of the super class Pisces typically showing gills. lean fish are those fish that store most of their fat in the liver.1. Fish flesh is dominated by the abundance of white muscle in relatively short segments. Covadonga Arias Department of Fisheries and Allied Aquacultures Auburn University Auburn Alabama 36849 United States of America 1. and excludes molluscs and Crustacea. while fatty fish have fat cells distributed along their bodies. most fish contain omega 3-fatty acids and other essential nutrients. CHILLED AND FROZEN RAW FISH Associate Prof. but typically present a low fat percent when compared to other meats. “fish” will be used for fresh and seawater finfish. it shows a typical season pattern especially in migratory species such as herring or mackerel. In addition. giving it its characteristically flaky structure. typically fluctuates between 0. Fish can be divided into fatty and lean fish. fins and a streamline body. ‘fish’ also refers to the flesh of such animals used as food. typically 3 and 15% of total weight. Muscle composition and structure of fish also differ from those found in other meat. However. 1 . and is mostly interspersed between the muscle fibres.2 .5% (1). In addition. and spawning season. Fish fillets are a poor source of carbohydrates. The connective tissue content of fish is also lower than that found in meat. Although fish is broadly similar in composition and structure to meat there are a number of distinctive features. The lipid fraction is the component showing the greatest variation.21%.20%. crustaceans. The lipid content. In this chapter. some regulatory agencies such as the US Food and Drug Administration (FDA) will include molluscan shellfish. which can be up to 67%.
Organoleptic refers to qualities such as appearance. Raw fish refers to fish that has not been cooked but excludes fish treated with curing salts and/or subjected to fermentation. but not below 3 °C during storage. and texture of the fish. transportation and sale. but no further control is exercised during storage. Spoilage describes changes that render fish objectionable to consumers.FISH AND SEAFOOD Chilled fish is fish that has been cooled to. and maintained at or below 7 °C. transportation and sale. DMA is dimethyl amine. usually carbon dioxide (CO2). and maintained at or below -2 °C (normally below -12 °C) during storage. Shelf life is defined as the time of storage before microbial spoilage of a fish is evident. Spoilage potential is a measure of the propensity of microorganisms to render fish objectionable to consumers through the production of offensive metabolic byproducts. 2 . renders that fish objectionable to consumers. colour. firmness. transportation and sale. marbling. Fresh fish is raw fish that has not been processed. Modified-atmosphere packaging (MAP) refers to packaging systems in which the natural gaseous environment around the product is intentionally replaced by other gases. and maintained at temperatures just below the freezing point. Superchilled fish is fish that has been cooled to. odour and texture. during storage. This technique is primarily used for bulk storage. Controlled-atmosphere packaging (CAP) refers to packaging in an atmosphere where the composition of the gases is continuously controlled during storage. spoilage microflora describes an association of microorganisms that. and flavour based on the maturity. Quality refers to palatability and organoleptic characteristics such as tenderness. colour. juiciness. nitrogen (N2) and oxygen (O2). Frozen fish is fish that has been cooled to. through their development on fish. Evisceration is the removal of the viscera from a fish. The proportion of each component is fixed when the mixture is introduced. at -2 to -4 °C. frozen or preserved. hence.
Flavobacterium.II) (3).CHILLED AND FROZEN RAW FISH Vacuum packaging (VP) refers to packaging systems in which the air is evacuated and the package sealed. and those that although not autochthonous to the aquatic environment. whenever possible. Lactobacillus and coryneforms can also be found in varying proportions (1). Human pathogenic bacteria can be part of the initial microflora of fish. Members of the families Vibrionaceae (Vibrio and Photobacterium) and the Aeromonadaceae (Aeromonas spp. since they heavily depend on the type of sample analysed and the protocol used for isolation. the natural fish microflora tends to reflect the microbial communities of the surrounding waters. The number and diversity of microbes associated with fish depend on the geographical location. microorganisms colonise the skin. standard culture-dependent methods can only recover between 1 to 10% of total bacteria present in any given sample. On the contrary. In general. molecular-based methods have not yet been used to address this issue. several culture media containing sodium chloride and more than one incubation temperature should be used.) are also common aquatic bacteria. More accurate. For example. It is crucial to mimic the environmental physicochemical parameters when isolating bacteria from fish. although these are influenced by water quality and feed. posing a concern for seafoodborne illnesses (2). Clostridium. The autochthonous bacterial flora of fish is dominated by Gram-negative genera including: Acinetobacter. as with other vertebrates. It must be noted that mesophilic bacteria can rapidly overgrow psychrotropic organisms. healthy fish is considered sterile and should not present any bacteria or other microorganisms. gills and the gastrointestinal tract of fish. are present there as result of contamination (anthropomorphic origin or other) or are introduced to the fish during harvest. Gastrointestinal tracts and gills typically yield high bacteria numbers. Fish harvested from clean and cold waters will present lower bacterial numbers than fish from eutrophic and/or warm waters. and typical of the fish flora. 3 .I). Shewanella and Pseudomonas. It is difficult to estimate how many microorganisms are typically associated with fish. the season and the method of harvest. In fact. These pathogens can be divided into two groups: organisms naturally present on fish (Table 1. potential human pathogens may be present in both scenarios. 1. Moraxella. Micrococcus.2 Initial Microflora The subsurface flesh of live. processing or storage (Table 1. some species (most Vibrios) require sodium chloride for growth. Gram-positive organisms such as Bacillus. However.
In addition.37 °C High for most species.14 . numbers of pathogenic bacteria in raw fish tend to be low. 105 .0.I Indigenous bacterial pathogens typically present in fish (3) Organism Temperature range Estimated Minimum Infective Dose 00.106 cells/g (exception: V. Intestinal track of terrestrial vertebrates Human origin Fecal contamination Ubiquitous in environment Shigella spp. 4).II Non-indigenous bacterial pathogens frequently present in fish (3) Organism Listeria monocytogenes* Primary habitat Soil.1 µg toxin lethal dose Clostridium botulinum non-proteolytic type E Pathogenic Vibrio spp.102 101 . human origin Minimum Infective Dose Variable depending on the strain (>102 cells/g) Staphylococcus aureus 105 . sewage.109 cells/g) * Some sources considered L. estuarine environments.37 °C Unknown Unknown TABLE 1. during storage indigenous spoilage bacteria tend to outgrow potential pathogenic bacteria. 4 .103 High (107 . However. Vibrio cholerae Vibrio parahaemolyticus Vibrio vulnificus other vibrios Aeromonas spp. stream water.19 µg/kg bodyweight from < 102 . Escherichia coli Yersinia enterocolitica 101 .1 .35 °C 8 . Toxin levels 0.FISH AND SEAFOOD TABLE 1. monocytogenes as part of the natural aquatic flora It is apparent from the above that there is potentially a very diverse range of organisms present on fish.106 cells/g. Plesiomonas shigelloides 3 . and risk associated with the consumption of seafood is low (2. vulnificus) 5 . and mud Ubiquitous. birds.>106 Salmonella spp.26 °C 10 .
freezing facilities (-18 °C) may be used to prevent the catch from deteriorating. care should be taken in removing all the gut contents and washing the carcass thoroughly prior to refrigerating. in recent decades. This contrasts greatly with the meat industry. After capture. however. human hands and clothing. and conditions of storage. ropes. potential contaminants added during handling and processing. The length of time that set nets have been in the water or the time trawlers’ nets are towed. In eviscerated fish.3 1. If evisceration is carried out at sea. 1. Physical damage such as loss of scales. offering better control of fish health prior to. and their re-use will increase the crosscontamination of other fish with such microorganisms.1 Processing and its Effects on the Microflora Capture. In addition. in which the health of each animal can be assessed prior to slaughter. The decision to eviscerate the catch at sea will depend greatly on the size of the fish and the duration of storage at sea. However. with fish such as tuna and cod being more commonly eviscerated than sardines. These contacts will not only increase the bacterial cross-contamination between fish batches but will introduce microorganisms from other sources such as humans. the fish may be stored in the vessel for periods ranging from just a few hours to several weeks in melting ice. and during harvest. the cuttings provide areas of exposed flesh that are open to microbial attack. and the killing process is designed to minimise stress. or traps. Fish may be eviscerated prior to storage at sea . Increasingly. in which the bacterial load 5 . with the production of off-odours. During capture and storage.3. Used refrigerated brines can be contaminated with high numbers of psychrotrophic spoilage bacteria. handling and processing Wild finfish are usually caught by net. and especially when fish is stored on board for longer periods. decks. digestion and off-flavours in uneviscerated fish. The action of intestinal enzymes and activity of the gut bacteria on the flesh around the belly cavity may produce discolouration. with very little control over the condition of the fish at the time of death or the duration of the killing process. birds and soil.a practice that may have both advantages and disadvantages. mackerel or herring. chilled brine or refrigerated seawater at -2 °C. hook and line. cortisone levels increase during prolonged stress and can alter the fillet quality. finfish will almost invariably come into contact with nets. boxes and/or baskets. Of particular concern is the use of wooden or soiled plastic containers for storage and unloading at the quayside. bruising and bursting of the gut will increase the number of sites open for bacterial attack and spread. Inadequate circulation of chilled brines may result in localised anaerobic growth of some microorganisms.CHILLED AND FROZEN RAW FISH Shelf life depends on the initial microflora on the fish. icing or freezing. has an effect on the amount of stress and physical damage that the fish will suffer during capture. aquaculture practices have been expanding worldwide. and spoilage.
and of the quality of the water used for ice and to wash fish. potential spoilage bacteria are among the psychrophilic and psychrotrophic flora present on temperate-water fish before death (9). Anaerobic atmospheres have less effect on fresh fish shelf life. Monitoring of the seawater for algal growth in order to limit the risk of algal toxin ingestion. use of effective detergents and disinfectants. fish have a higher post mortem pH.FISH AND SEAFOOD can be substantial. but if applied properly should extend the life of the product. oxygen etc. Carbon dioxide is the most important gas in MAP of fish because of its bacteriostatic and fungistatic properties. Both carbon dioxide and low pH inhibit the growth of the typical spoilage bacteria such as Pseudomonas and Shewanella. cleaning of the work environment. Bacterial composition under MAP shifts from mostly Gram-negative to predominantly Gram-positive (lactic) bacteria. in time. Low oxygen levels have been shown to substantially prolong the freshness and quality life of refrigerated seafood products. 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. In MAP. often in the absence of adequate refrigeration. In the absence of oxygen. not part of the normal flora of the meat animals (8). 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). the natural atmosphere is replaced with a controlled gas mixture (carbon dioxide.2 Modified-atmosphere packaging A natural atmosphere rich in oxygen (21%) is responsible for oxidative processes and for all aerobic respiratory life. The atmospheric conditions surrounding a product may be passively or actively altered. By vacuum packing foods. partial fermentation of sugars occur leading to lower pH. a reduction in the oxygen tension is achieved which. careful and sanitary handling during processing is required to reduce the risk of contamination with potential human pathogens. but isolate the food in an environment different from the bulk atmosphere. 1.3. However. These changes can be deleterious. What is more. As with all foods. if there is some oxygen demand from the product. Physical barriers not only protect from physical damage. 7). Packaging changes the intrinsic and extrinsic parameters affecting a product.). These containers are also used for displaying the fish during auction at the quayside. nitrogen. for example. however. from water activity (aw) through to physical damage. Brochothrix thermosphacta and psychrotrophic lactic acid bacteria (LAB) can produce spoilage characteristics. MAP extends the shelf life of most fishery products by inhibiting bacterial growth and autoxidation. will result in fully anoxic conditions. they are usually process contaminants. allowing more growth of spoilage organisms. and minimising handling will all reduce microbial cross-contamination. by actively altering the composition of the 6 . ferric ion (Fe3+)).
the National Advisory Committee on Microbiological Criteria for Foods (NACMCF) has published a number of recommendations on 7 . but can dissolve into fish liquids and deform packages. gutted or filleted fish. may not be considered ‘fresh’ fish. VP is one of the oldest forms of altering the interior gaseous environment of a pack. Flavobacterium. Replacement of oxygen with nitrogen. Of particular concern are the psychrotrophic type E and non-proteolytic type B and F strains. which provides guidance relating to vacuum or modified-atmosphere packaging for specific fish products (14). the higher pH and reduction potential (Eh) of the fish muscle. nitrous oxide and carbon monoxide have also been suggested as possible replacement gases in trace amounts for MAP/CAP. slower-growing flora (11). although less information on their effectiveness is available. The products. and the problems of muscle structure damage by the modified-atmosphere (11). The single most important concern with respect to the use of MAP is the potential for outgrowth and toxin production by C. Growth and toxin production have been detected in artificially contaminated packs of whole trout after 1 week’s incubation at 10 °C (13). The (International) Codex Committee has published a Code of Practice for Fish and Fishery Products (CAC/RCP 52-2003). The principal effect of raised carbon dioxide-MAP is an extension of the 'lag' phase of the growth of the bacteria on the fish. In the United States. The microflora of meat is not the same as that of whole. a modified-atmosphere may contain any gas necessary for the desired effect. Sulphur dioxide.CHILLED AND FROZEN RAW FISH surrounding gas. and the promotion of a predominantly Gram-positive. MAP has been applied to fresh meat and fish with a resulting commercially viable extension in shelf life (10). This has been driven partly by increased consumer demand for fresh and chilled convenience foods containing fewer chemical preservatives. including salt. Gases used in MAP of fish most commonly include carbon dioxide and nitrogen. the inhibition of common 'spoilage' bacteria (Pseudomonas. and the MAP of fish has more challenges to overcome as a result of: a comparatively large initial load of bacteria present.3 °C and produce toxins. High concentrations of carbon dioxide have the most pronounced microbial effect. without overt signs of spoilage. which are able to grow rapidly at low temperature. as they are able to grow at temperatures as low as 3. and increase in-pack drip (12). phosphates. Modified-atmosphere preservation of fish was first reported in the 1930s. Micrococcus and Moraxella). after additive application. an inert and odourless gas. sorbates and chelating agents such as ethylenediaminetetraacetic acid (EDTA) (11). does inhibit some aerobic bacteria and reduce the rate of oxidative rancidity. Many additives have been tried in conjunction with changed atmosphere. the pathogens that may be able to grow before spoilage occurs. discolour pigmented fish (11). but only in recent years has it seen a marked expansion in use and market share. but residual oxygen and other electron acceptors may be sufficient to allow oxidative spoilage of fish (12). botulinum.
use of high-quality raw fish. 5. from both a microbiological and a chemical standpoint. botulinum. grouping of fish types is possible to estimate the best carbon dioxide level to be used in MAP. 3. Only the highest-quality fish. These conditions include storage of the product at ≤ 3. The growth of these bacteria is reduced by high carbon dioxide concentrations but. Freshwater fish have different intrinsic bacterial flora and also do not contain such high concentrations of amines (e. 4.3 °C at all points from packaging onwards. however.g. Although all fish species need to be treated separately. It was also noted that organoleptic spoilage and rejection by the consumer should occur before the possibility of toxin production. TMAO) that can be reduced to produce 8 . 2. should be used. 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). greater advantages are seen. The addition of high levels of carbon dioxide to these fish. the Sea Fish Industry Authority has published guidelines for the handling of fish packaged in a controlled-atmosphere (16). Sensitive and accurate time-temperature indicators must be used to ensure that the product has not been temperature-abused. trimethylamine and sulfides). It highlights temperature control as the primary preventive measure against the possible hazard of toxin production by C. For fatty fish. as already stated. 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. Subsequent storage must be at or below 3 °C. owing to the dissolution of cellular structures. rejection due to alteration in fish texture with greater carbon dioxide makes this unsuitable (13).g. leading to the recommendation that the sale of MAP/VP raw fishery products be allowed only when certain conditions are met. from where drip loss and gaping of the muscle meant that organoleptically classified rejection occurred sooner than with no carbon dioxide addition (12). this problem occurs to a lesser extent with meat. adequate product labelling in respect of storage temperature. may result in unacceptable colour and textural change and hence little shelf life extension (11). oxidative rancidity is a significant source of spoilage and hence the complete removal of oxygen and the use of oxygen-impermeable films are preferred. at all times. but is recognised as a drawback of carbon dioxide application. adequate shelf life and cooking requirements. The replacement gas should contain at least 40% carbon dioxide. and the use of a HACCP plan. The 1985 British Guidelines on MAP/CAP fish from the National Fisheries Institute state that: 1.FISH AND SEAFOOD the safety of MAP and VP refrigerated raw fishery products (15). The problem of water loss in MAP fish is common. however. The only processing dip to be used is 5% potassium sorbate and 10% sodium tripolyphosphate. For other less fatty marine fish. In the United Kingdom.
CHILLED AND FROZEN RAW FISH off-odours. but it is a commonly held view that loss of freshness is related to autolytic degradation and spoilage is more microbial in origin (1). Taste panel evaluation of cooked fish may be used to score or grade colour. Over time. When working with fillets.60% carbon dioxide with a balance of nitrogen for fatty fish have been recommended (16. Loss of freshness and spoilage cannot be separated as processes.III. and this pattern is the basis of freshness grading schemes. with no smell. loss of the fresh characteristics may be simply measured by comparative visual and smell analysis. Various methods for whole fish freshness evaluation by a trained panel have been proposed. and the flesh turns from bright and elastic to dull and soft (1). 1. these are summarised in Table 1. Gas mixtures of 40% CO2/30% N2/30% O2 for white fish and 40 . so lower carbon dioxide concentrations may have better preservative effects. The eyes turn from convex and clear to concave and opaque. and metabolic activities of microorganisms present in the fish. or for that matter evaluation of the taste of the fish. texture. the gills from pink and shiny. A. 17.4. 18) and are probably the most widely used. Spoilage of fish starts upon death due to autoxidation (oxidation of unsaturated lipids).4 1. evaluation of taste is unavoidable as the visual pattern of degradation of the eyes and gills is unavailable for analysis.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. smell and 9 . rejection of whole fresh fish for human consumption may be made without chemical or microbiological evaluation. as traditional spoilage patterns of the external organs are typically very distinct. C (C=reject) Significantly. TABLE 1. to brown and slimy with an intense off-odour. reactions caused by activities of the fish’s own enzymes. Degradation of whole fresh fish stored in ice generally follows a set pattern. B. the skin turns from iridescent to dull and bleached with bacterial slime.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).
Because fish tend to have relatively little residual glycogen compared with mammals. iv) Enhancement of the spoilage characteristic of phase iii. spoiled and putrid. To a varying extent. TMAO permits a high Eh to remain in the muscle tissue. the muscles harden as residual adenosine triphosphate (ATP) is reduced and the myosin and actin filaments bind to form actomyosin (18). The processes of degradation being analysed by the quality scoring methods above are a complex mix of physical.FISH AND SEAFOOD taste of cooked fish. sea-weedy taste. post-rigor values are typically in the range of pH 5. chemical. The process of ATP breakdown is used as an indicator of fish freshness (24). this will generally correlate with the length and severity of rigor. which has been ascribed a number of possible functions as: a trimethylamine (TMA) detoxified waste product. little flavour. 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. the cooked analysis of fish passes through four phases of spoilage: i) Delicate sweet.8 to 6. The pH of the muscle will drop after death depending upon the amount of residual glucose or glycogen that is reduced anaerobically to lactate with the coproduction of ATP. which increases from 0% towards 100% with time. an osmoregulator. After death. These processes are strongly influenced by the physical conditions of storage.g. fruity and/or bitter off-flavours. reduction in ATP and increase 10 . as little endogenous reduction of TMAO occurs (18). sulphurous and/or rancid smells. %). all marine fish use/have TMAO (21). possibly slightly metallic. a ratio of concentration of the hypoxanthine (Hx) and inosine (HxR) to total concentration (ATP. HxR and Hx) gives a quantity (K-value. is significant and may even be responsible for the ultimate sensory rejection of fresh cod and other fish with high initial TMAO content (20. the pH drop of the muscle is correspondingly less. biochemical and microbiological actions. the Eh of fish muscle remains relatively high (20). From being flaccid. the conductance generally increases. an anti-freeze. iii) Traces of sour. bacterial reduction of TMAO to TMA. 23). texture becomes soft and watery or hard and dry. rigor resolves. By measuring the concentrations of the six components. Generally. RT Freshmeter or Fishtester). As the fish degrades. During the initial storage. rigor mortis is the first noticeable change in the fish. IMP. However. ADP. ii) Neutral taste. or its remaining shelf life (1). 19).5 (18. development of sickly sweet. ammoniacal. thus. the muscles relax again and the fish returns to a flaccid state. cabbage-like. or simply a waste product present due to bioaccumulation (22). After death. AMP. 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. After some time. the Torrymeter. an intense odour compound.
or a specific fraction of the volatile bases. however. 31). colour changes.CHILLED AND FROZEN RAW FISH in hypoxanthine by endogenous enzymes allow this measurement to be used for determining freshness (19). During spoilage. therefore. However. The Conway method (and variations of it) uses a strong inorganic base to volatilise the bases in the fish sample. not to describe freshness. is later significantly reduced by bacterial action. the importance of microbial action increases with time (5. however. 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. a correlation can be made between the spoilage of the fresh fish and the production of TVB-N. This measure is highly dependent upon the temperature of storage and may not reflect the rate of loss of quality equally at different temperatures. Lipid oxidation can be a result of enzymic action or a cascade reaction initiated by free radicals (26. Evaluation of Total Volatile Base-Nitrogen (TVB-N. also termed TVN). does not show correlation between muscle pH and the amount of volatile bases contained within the fish at rejection. 26). as microbial spoilage is limited by the low storage temperature (1. 27) produced by aerobic respiration and other forms of metal ion reduction (28). 11 . 7). as significant proteolysis is observed only during the latest stages of spoilage and after rejection (30. and are especially important in the spoilage of frozen fish. which may also be formed by bacteria (25). and a segregated weak acid to absorb them. using Conway diffusion chambers (29) allows determination of changes of mg-N/100 g fish. the majority of volatile bases are produced from the soluble non-protein nitrogen of the fish (free amino acids and other low-molecular-weight nitrogenous compounds). hypoxanthine. Lipid oxidation and other oxidative changes lead to oxidative rancidity. for example the TMA fraction. The major spoilage odours and flavours of fresh fish are undoubtedly principally microbial in origin. For some fish species. Comparing results for different fish species. Chemical degradation continues after the initial post mortem phase. Volatile bases are the best-characterised chemical indicators of fresh fish spoilage. 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. so the measurement is effective only during the initial loss of freshness. Quality indices based upon the products of microbial metabolism do not explain changes in quality until microbial growth produces measurable changes in the fish. the residual acid is then titrated. these measurements are usually used to quantify the amount of spoilage. Oxidative rancidity is known to reduce the quality of fatty fish in particular (19. Neither does the change in pH during storage correlate well with the production of TVB-N. 18).
5. The sum of these changes may result in a different spoilage profile. biochemical and microbiological reactions taking place within and on the fish. the lower the temperature of storage. careful temperature control during storage is not only important in terms of quality loss. 5). by altering the gas atmosphere (extrinsic). but also crucial for the assurance of consumer safety. The factors are interactive and cannot be completely isolated. a minority are often associated with the majority of the spoilage. 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 1. For fresh fish.1 Factors Affecting Fresh Fish Spoilage Temperature By far the most effective method of reducing the rate of whole fresh fish spoilage is temperature control (1. All chemical reaction rate kinetics (and thereby microbial growth) are temperature-dependent. 34). 1.and temperate-water fish spoilage rates may be many.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). Not all the bacteria growing on a food will lead to the production of objectionable characteristics. For fresh fish iced immediately after capture and continually stored in ice. 1. the slower the spoilage processes proceed (within limits). Also.5. This increase has been reported to be moderately dependent on the temperature of the sea from which the fish are taken (33. Fish spoil as a result of the chemical. For example. the pH of the fish muscle may change (intrinsic) and a different microbial population may develop due to the change in atmosphere and pH.FISH AND SEAFOOD 1. including the ability of the endogenous bacteria to grow at low temperatures and lower endogenous enzyme activity.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. realisation that the bulk of the microbial population on newly caught fish does not cause off-flavours and off-odours stems from work 12 . The concept of specific spoilage organisms (SSOs) is not new.5. yoghurt spoilage by yeasts and clostridial spoilage of cheese (5) are examples where it has been recognised for many years that a particular minority of the microbial flora present in the product is responsible for its spoilage. the results of altering one parameter may have far-reaching consequences on others. The reasons for the differences between tropical.
oxidase-positive. It is also present in fresh water. work with inoculated sterile fish blocks resulted in identification of a specific minority of microorganisms. It is thus an important spoilage organism of gadoid fish such as cod. It is able to grow as fast as. esters and nonhydrogen sulphide sulphur-containing compounds such as methyl sulphide (36. These organisms are described as potential spoilers. 13 . The Pseudomonadaceae family represent a large and poorly defined group of microorganisms. frozen) fish products. which produced the characteristic spoilage compounds of the fish (30.5. As previously noted. Sh.3. the characterisation of spoilage of each product must be made before the identification of the responsible agent(s) can proceed.1 Pseudomonas spp.10% of the total flora of fresh fish from temperate marine waters. plus other characteristic compounds of the species responsible for spoilage.2 Shewanella putrefaciens Sh. mediated spoilage is characterised by ‘fruity’. 1. putrefaciens growth may be used as indicators of spoilage (30). Bacteria identified as being associated with the spoilage process of fresh fish are as follows: 1. 37). ‘oniony’ and ‘faecal’ odours from the production of ketones. 35). on fish are diverse and in many cases species-specific. it is often isolated as about 1 . or faster than the rest of the flora of ice-stored fresh marine fish (9). its ability to reduce TMAO to TMA.3. putrefaciens is considered an SSO of temperate-water marine fish species stored in ice. catalase-positive. 35. and proteolytic and lipolytic enzymes that may affect the quality of fresh and. 36). processed (e. i. The spoilage compounds associated with the growth of psychrotrophic Pseudomonas spp. therefore. form methylmercaptane (CH3SH) and dimethylsuphide ((CH3)2S) from methionine and produce hypoxanthine (Hx) from inosine monophosphate (IMP) or inosine. produce hydrogen sulphide (H2S) from cysteine. aldehydes. obligate respiratory bacteria.CHILLED AND FROZEN RAW FISH started in the 1940s.5. The importance of Sh. more especially.e.g. putrefaciens spoilage of fish is due to its biochemical action on muscle. The spoilage of freshwater fish is generally ascribed to the growth of Pseudomonas spp. but only if or when they reach numbers capable of producing sufficient spoilage compounds to effect rejection do they become the SSOs of the product (9. putrefaciens to the spoilage of fresh marine fish has been recognised since the 1940s. During the 1970s. They are generally characterised as Gram-negative rods. the spoilage of a product may be strongly influenced by the conditions under which the product is held. although only since the late 1960s has it been realised that specific metabolites of Sh. motile with polar flagella. and may play some role in the spoilage of freshwater fish. Members of the genus are able to produce pigments. (9) and they are considered SSO of iced freshwater fish (35). Pseudomonas spp.
as the ferric iron is insoluble.how to reduce a molecule that they are not able to take into their cells efficiently. putrefaciens strains are characterised as microaerophiles or anaerobes. it was proposed that this organism. putrefaciens growth has been collected (12. This wide-ranging respiratory capability is thought to be unique (39). like all members of the Pseudomonadaceae. owing to its cell size and activity. putrefaciens lies in its chemotaxis: the bacterium. and Shewanella aquimarina. and its metal ion reduction ability has led to intensive investigation into its role in iron and sulphur cycles. Shewanella waksmanii. unlike many other motile bacteria. In 50% N2/50% CO2 MAP cod. In terms of its taxonomy. putrefaciens were identified as Shewanella algae. 1. a population of Ph. although. phosphoreum increased in notoriety when it was proposed that reduction of TMAO to TMA limited the shelf life of MAP cod fillets (12). they are heterogeneous. but does show strong chemotaxis up gradients of most of their alternative electron acceptors (39). Shewanella denitrificans. at the time of rejection. additional psychrotrophic species such as Shewanalla baltica. putrefaciens were required. Shewanella oneidensis. the bacteria have a significant problem to overcome . phosphoreum were required for 50% of taste panellists to reject a sample. Shewanella frigidimarina. The prevalence of the bacterium in so many environments. Research has shown that approximately 107 cfu/g of Ph. mesophilic strains of Sh. Shewanella gelidimarina. 38). Because no other TMAO-reducing bacteria were present in sufficient numbers to produce the quantities of TMA that were related to rejection. and Shewanella profunda were described. Shewanella livingstonensis.FISH AND SEAFOOD for which the most compelling evidence showing spoilage as a result of Sh. putrefaciens was not present in such numbers (23. does not show carbon-source chemotaxis. 41). phosphoreum sufficient to cause spoilage was found. Shewanella affinis. The localisation of the reductase on the outer cellular membrane offers the bacteria a solution to this problem (40). 41). Another distinct feature of the physiology of Sh. putrefaciens may grow in the absence of oxygen using alternative terminal electron acceptors (ATECs). but Sh. with several new species being described. Ph. Ph. Later. It is also able to produce hydrogen sulphide and a range of other off-odour compounds.5. This type of intoxication occurs when bacteria convert the histidine 14 . Sh. baltica has been identified as the main H2S producer in cod during cold storing (37). In fact. putrefaciens has been studied.3 Photobacterium phosphoreum Recognised for some time as being present on spoiling fish (30).3. Shewanella olleyana. Initially. whereas >108 cfu/g Sh. it is strictly respiratory. The ferric reductase activity of Sh. Sh. phosphoreum has also been identified as responsible for histamine fish poisoning. Sh. was capable of being in a significant numerical minority on the MAP fish but still able to yield the majority of the TMA thought to be responsible for the rejection (23. and recent taxonomic description using modern molecular methods bifurcated this species.
Fewer still are capable of growing on fresh fish. Of these. C1. The heterogeneity of the types (A. but with proper cooking. these risks are substantially reduced (1.e. 5). thermosphacta is a well-characterised psychrophilic spoilage organism of meat. E and F include bacteria with minimum growth temperatures of approximately 3. F and G) is well documented (44). E.4 Brochothrix thermosphacta and lactic acid bacteria B.6. 1. 5) reports that fresh fish consumption has never been shown to cause human botulism. lightly preserved fish products are associated with botulism. a neurotoxin that causes flaccid paralysis and is associated with a variable (44) mortality depending upon dose. presents a potential risk. Types A. Huss (1.CHILLED AND FROZEN RAW FISH present in fish into histamine. including antisera. thermosphacta on spoiling fish in a 40% CO2/30% N2/30% O2 MAP (42).5. B. Growing evidence suggests a role for B. bacteria and higher parasites all present some risk associated with fish consumption (1. where a particular 15 . it is bacterial risks that increase after capture of the fish. especially type E botulinum intoxication.6 Pathogens: Growth and Survival Endogenous chemicals. However. a convenient but diverse species of bacterium with a number of different types. thermosphacta did not produce off-flavours until 2 .3 days after having reached 108 cfu/g (42). 1. C2. B. 6). MAP studies have also demonstrated this organism’s sensitivity to oxygen and have shown that they are also inhibited by high CO2 concentration (43). E.3. Some Ph. Acetate production has been reported as a good indicator of spoilage by this organism. B. 5. When inoculated into VP corned beef and sliced ham. D. previous exposure and access to supportive treatment. Review of outbreak data suggests fresh fish to be safe. Of 404 intoxication outbreaks of type E C. botulinum recorded up to 1963. algal toxins. they do not produce significant product spoilage). this is probably due to spoilage occurring before toxin elaboration. age. 1. botulinum. 75% (with a 35% mortality rate) occurred in Japan. There are few human bacterial pathogens that can cause primary infections or disease and are capable of persisting in the aquatic environment. The remaining few bacteria present a major risk involved with the consumption of raw seafood such as sushi or oysters. types B.1 Clostridium botulinum C. phosphoreum strains have great capacity as histamine producers even under refrigeration conditions. Recent studies have investigated the dominance of B. (and very rarely F and G) have been reported to cause human disease. human viruses. thermosphacta in the spoilage of some MAP fish.3 °C (Group II) and these are non-proteolytic (i. Adult disease is caused by the production of the botulinum toxin.
and proper cooking (boiling for 1 minute. Proper chilling. again. which is believed to be required for virulence. parahaemolyticus infections since 1996 and is referred to as the pandemic strain (47). Since 1996. 11).2005). Only about 1% of marine isolates produce a thermostable haemolysin. France. storage and preparation (11). USA. or cooking at 80 °C for 5 minutes) (44) will substantially reduce risk. parahaemolyticus is a marine organism. which was probably contaminated from an uncooked food source or an excreting foodhandler. is small. use of post harvest treatments and/or cooking of fresh seafood will reduce risks. the majority are associated with products not cooked immediately before consumption. Human disease caused by the organism has only been observed in conjunction with marine bivalve (mostly oysters) and some crustacean consumption. only shellfish are associated with the disease and no reported cases by the Center for Diseases Control. accounted for the majority of cases (1. due to the cultural preference for raw fish dishes. Currently. were associated with finfish between 1978 and 1998 (46). parahaemolyticus in sea water fish and shellfish that could potentially be hazardous to humans. there is no specific guideline that describes a minimum level of V. Other lightly or semi-preserved (e. 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. including epidemics in India. V. A unique clone of V. there remains only an ‘extremely small’ risk of intoxication if reasonable precautions are taken during handling. 1978 . Despite the ubiquitous nature of type E C. which can cause human gastroenteritis.g. and North America. vulnificus is associated with warm marine and estuarine waters.2 Vibrio parahaemolyticus and other vibrios V. smoked. salted or pickled) fish have also been associated with botulism. The risk of toxin formation before apparent spoilage has been studied for some MAP fish (45). Proper temperature control (<3. Russia. Generally. especially in individuals with underlying diseases (patients suffer from immune and liver diseases or blood 16 . This strain has been responsible for 50 to 80% of all V. The first report of the organism being a foodborne agent was with shirasu.g. parahaemolyticus outbreaks.FISH AND SEAFOOD fermented raw fish product that is consumed uncooked.3 °C) will eliminate all risk. parahaemolyticus O3:K6 is responsible for many of the recent V. Southeast Asia. sushi) (5). The bacterium causes primary septicaemia. In Japan. botulinum in the marine environment (isolated from around 90% of marine and environmental samples from northern European waters and its consequent presence in seafood (a surveyed incidence of as high as 65%) (2. parahaemolyticus cases have increased across the world. 5). 1. the majority of outbreaks are caused by consumption of raw fish products (e. whether stored aerobically or in modified-atmospheres. but as fresh fish are often cooked before consumption and the botulinum toxin is heat-labile. a Japanese boiled and semi-dried sardine dish. the risk posed to the consumer of fresh fish.6. V. Japan. I-sushi. and when in contact with contaminated water (no CDC reported cases from fish.
Despite this. Again. L. Although the psychrophiles are unable to grow at body temperature. V. sodium channel blocking and haemolytic activity (48). shigelloides is similar to Aeromonas spp. with minimum growth temperatures around 0 °C and optima of 15 . 5. possibly due to under-reporting because of their likeness to E. Some isolates are true psychrophiles. 1. there is no risk from the consumption of properly handled and cooked fresh fish.6. the causative agent of cholera. but the bacterium is known to survive and grow in the shallow marine. The bacterium has been documented as being responsible for a few fishborne outbreaks of gastroenteritis. There is no risk associated with the consumption of properly chilled and cooked fresh fish. is historically associated with faecally contaminated water. but proper cooking should substantially reduce risk of infection. and especially Aeromonas hydrophila. cholerae O1. coli on isolation media (50). are present on fish and can grow to significant numbers during storage (1.000 17 . In the US. proper chill storage will eliminate any risk associated with this organism.55 cases per 100. which is often fatal (>50%). others are psychrotrophic mesophiles (49). listeriosis cases have been reportable to the CDC.26 to 0. in its habitat.3 Aeromonas Aeromonas spp. and proper cooking will substantially reduce risk. no more than circumstantial evidence exists linking the consumption of seafood to Aeromonas spp.CHILLED AND FROZEN RAW FISH disorders). monocytogenes has been well documented as a foodborne human pathogen. 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. and especially estuarine environment. It is known that 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. They are aquatic organisms with an epidemiology that is yet to be fully understood. Since 2000. shigelloides are encountered. The major virulence factor appears to be the production of toxins with enterotoxic. Most cases have been sporadic rather than associated to large outbreaks. It is particularly associated with disease following consumption of raw oysters from warm sewagepolluted waters. infection. and fish and shellfish are probably the major reservoirs for the organism (50). are associated with human diarrhoeal illnesses. 1.6.4 Listeria monocytogenes L. Chill storage may not eliminate growth. monocytogenes incidence has been between 0. Similar problems as those encountered for Aeromonas spp.. in assessing the risk of fishborne infection by P. cytotoxic. however. their potential to produce disease-causing exotoxin in food has not been fully examined (48). P. 17).20 °C.
. scombroid poisoning is usually a mild intoxication and it is not a significant cause of death. 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. Decarboxylation of histidine by a wide range of bacteria including Enterobacteriacae. but more especially by Morganella morganii. AIDS patients and patients undergoing immunosuppressive therapy) and a lack of data covering infectious doses. Being a psychrotroph.. Photobacterium spp. If fish are not properly refrigerated.. 1.5 Scombroid fish poisoning The production of histamine and other biogenic amines with human immunological activity.pregnant women. it is not apparent to the consumer and it cannot be destroyed by cooking. 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. Infections are mostly associated with the consumption of raw or 18 . Owing to the dependence of the virulence of this organism on the immune state of the host (usually requiring lowered cellular immunity.6 Parasites Helminthic parasites can occur extensively in finfish but very few are capable of infecting humans. It is regularly isolated from seafoods (51). some Vibrio spp. within high histidine-containing fish (especially members of the Scombridae and Scomberesocidae families) is responsible for an intoxication termed scombroid poisoning (54). full risk analysis is not possible (52). Fortunately.FISH AND SEAFOOD persons.48 minutes in salmon have been reported (53). Low-temperature storage of potentially toxigenic (<5 °C) fish at all times is the most effective way to control histamine production (6). 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. Of fish-transmitted human diseases scombroid poisoning is commonly reported in the US (118 outbreaks with 463 cases reported by CDC 1998 . Clostridium spp. Larvae of parasites such as Anisakis simplex are resistant to curing and marinating but can be easily destroyed by freezing at -17 to -20 °C for 2 hrs. at risk groups . but cooking will significantly reduce the risk of infection. leads to the production of histamine. It is environmentally ubiquitous but its true frequency in the marine environment is not well studied.6. and Lactobacillus spp. Lightly preserved products. there is a far greater risk of unacceptable histamine levels being attained before sensory rejection. 1. but proper cooking will reduce any risk.2002 (55)) and is also common worldwide.6. foetuses.98 minutes in cod and D60 of 4. neonates. especially pickled fish. D60 of 1. alcoholics. Klebsiella pneumoniae and Hafnia alvei. 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. Like other toxins.
EC.IV) (57). A limited number of nematodes and trematodes found in finfish have also been identified as a cause of human disease (56).7 Published Microbiological Criteria Microbiological criteria for food defines the acceptability of a product or a food lot based on the absence or presence or number of microorganisms. TABLE1. Rome.CHILLED AND FROZEN RAW FISH mildly processed fish such as sushi. risk of human disease caused by natural bacterial contamination (not through faecal pollution) of fresh fish is extremely low. coli counts where this method is preferred. area or lot (CAC. 2003. For fish from inshore or inland waters of doubtful microbiological quality. 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.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. Gram L. FAO. An increase of APC to levels in excess of 106 cfu/g is usually indicative of inadequate refrigeration. Testing methods should be practical and the enforcement of such criteria should translate as a reduction of potential microbiological risks to consumers.1997. especially in warm-water areas and where fish are to be consumed raw. parahaemolyticus (see Table 1.. The most effective control of all bacterial risks is continual chill storage in ice (0 °C) or freezing. 444. long storage under refrigeration or one of the former prior to freezing. 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. In summary. Ababouch L. 1997) Huss H. coli proposed by the International Commission on Microbiological Specifications for Foods (ICMSF). 1.. including parasites and/or quantity of their toxins / metabolites per unit of mass. No. which in many cases will eliminate the risk. parahaemolyticus Staph. volume. aureus 10 7 7 2 3 3 19 5 5 5 0 2 2 0 102 103 103 104 . it may be desirable to test for Salmonella and V. Cooking properly immediately before consumption is also an effective way of reducing or eliminating risk of fresh fishborne disease. Criteria should set standards that are attainable by the currently accepted GMP and applied only when there is absolute need for it.H. Assessment and management of seafood safety and quality FAO Fisheries Technical Paper. Faecal coliform counts may be used instead of E.
1996. in FAO Fisheries Technical Paper No. Jeksrud W.J. M. Feldhusen F. 283-94.J. Codex Alimentarius Commission. Ed.R. Gram L. 1989.E. Kautter D. 121-37. Rome. 12. Ahmed F. 1994.S. Nychas G. Incidence and growth of psychrotrophic Clostridium botulinum in foods.H.T. Gram L.FISH AND SEAFOOD 1. 1995.H. 37. 1992. Food and Agriculture Organisation of the United Nations. 33.A. Jehanno D. Huss H. Food Control. 14.. Rhodehamel E. Gibbs P. parasites. Spoilage and shelf life extension of fresh fish and shell-fish. 27. 8 (2)..E. Davies A. Food Control.. Incidence of foodborne pathogens in European fish. 2002.. Codex Alimentarius Commission. 87121.N. 4. 6. Theoretical analysis of two hot water cabinet systems for decontamination of sides of beef.A. Rome. 5.R. 1651-60. Code of Practice for Fish and Fishery Products (CAC/RCP 52-2003). in FAO Fisheries Technical Paper No. 12. 2.H. 91-8. Microbes and Infection. Fletcher R. 19. 1992. Simpson B. 2003.. Food and Agriculture Organisation.. Assurance of seafood quality. 11. 87-118. 12. Huss H. Critical Reviews in Food Science and Nutrition. Control of indigenous pathogenic bacterial in seafood. Food and Agriculture Organisation of the United Nations. The role of seafood in bacterial foodborne diseases.K. International Journal of Food Microbiology. and natural toxins. 334.H. 5-7. Davies A.K. 243-60.. 7. References Huss H.R.8 1.H. activities and safety. Italy. 1994. Ashie I. Quality and quality changes in fresh fish.. Armstrong D. Reddy N. A review of modified atmosphere packaging of fish and fishery products – significance of microbial growth.. 5 (1). 1993. 2001. Spoilage and shelf life of cod fillets packed in vacuum or modified atmospheres. Food Control. Kirby R. Rosnes J. Journal of Food Safety. Ed. International Journal of Food Microbiology. 13. Davey K. International Journal of Food Sciences and Techonology. 9. 2000. 291-304. 107-27. Microbiological spoilage of fish and fish products. 1996.J. Dalgaard P.R. Huss H..P. Capell C... 8. 10. 36. International Journal of Food Microbiology. 348. Huss H.. 3.. Sivertsvik M.A. 2. Smith J. Shelf life extension and safety concerns about fresh fishery products packaged under modified atmospheres: a review. 20 . International Journal of Food Science and Technology. 67-71. Food and Agriculture Organisation. 24... Review: Assessing and managing risk due to consumption of seafood contaminated with microorganisms. 1997.
.E. 18. AVI Publishing. Hebard C. 1995. 1982.A. Edinburgh. 24.N. Saito T.ucdavis. 42-54.edu/haccp/compendium/chapt08. 1996.. Lipid oxidation in seafood. Davies A... Technomic. Eds. International Journal of Food Microbiology.. 1933. Ed. 1995. 354-8.E. Guidelines for the Handling of Fish Packed in a Controlled Atmosphere. Westport. 33.R. Biochemical Journal. Autolytic changes in fish post mortem.. 26. International Journal of Food Microbiology. Dalgaard P. 1995. .spoilage of fish. 1987. International Journal of Food Microbiology.M. 17. Lancaster. in Chemistry and Biology of Marine Food Products.htm Anon. Rome. 1. Ed. 26.J. 21. Schwall D.. Microbiological spoilage of fish and fish products. 20. Chemical composition. Part I . Flick G.. http://seafood. Qualitative and quantitative characterisation of spoilage bacteria from packed fish. 22. 1992..M. Flick G. 1985. 30.M. Rome. 117-27. Food and Agricultural Organisation of the United Nations.G. Surette M. Journal of Food Quality. 19-22.E. Huss H.. Changes in purine nucleotide of red lateral muscle of rainbow trout. 1995. Hebard C.. Food Technology.H. Washington D. 1992.H.. 245-67.J. Quality and Quality Changes. Ratnayake W.E. 25. Martin R. Gram L. Reginstein J. Børresen T.J. Eds. National Advisory Committee on Microbiological Criteria for Foods. 1971. Vacuum or Modified Atmosphere Packaging for Refrigerated Raw Fishery Products. 27. 23. Food and Agricultural Organisation of the United Nations. Letters in Applied Microbiology.. 21. Slade A. Biochemical basis of post-mortem nucleotide catabolism in cod (Gadus morhua) and its relationship to spoilage.E. 19.. in Fresh Fish. 41-50. Ackman R. Sea Fish Industry Authority. Shelf life extension of fresh fish . Non-enzymatic oxidation of seafood lipids. 1983. National Advisory Committee on Microbiological Criteria for Foods.. 27. An absorption apparatus for the micro-determination of certain volatile substances.. in Fresh Fish. Adopted. 1988. 2007. Conway E.J.a review. Khayat A. 29. 34.J. Fate of Aeromonas and Yersinia on modified-atmospherepackaged (MAP) cod and trout. 419-29.J.. Bacteria active in the spoilage of certain seafoods.A. Pedrosa-Menabrito A. 305-17. Hendrie M.S. Byrne A. March 20. 1-18. Herbert R. 21 16. Journal of Applied Bacteriology. Leblanc P. Quality and Quality Changes.H. The micro-determination of ammonia.CHILLED AND FROZEN RAW FISH 15. Nature. Arai K. 1959. Huis in't Veld J.J. Journal of Agricultural and Food Chemistry. Martin R. 11. Gill T.M. 28. 184.E. 36. Martin R. Flick G. 1996.H. in Advances in Seafood Biochemistry. Shewan J. Gill T.J. Microbial and biochemical spoilage of foods: an overview. Yajima T. 33. 121-37. Huss H. Huss H.C. Occurrence and significance of trimethylamine oxide and its derivatives in fish and shellfish. 130-2. Gibson D. 22-36. 1415.. 37.
The effect of storage temperature on shelf life. Rome. Pseudomonas fluorescens. 1992. Rome. 37. 26. Myers C. Volatile compounds produced in sterile fish muscle (Sebastes melanops) by Pseudomonas putrefaciens. Identification of Shewanella baltica as the most important H2S-producing species during iced storage of Danish marine fish. Scanlan R. COUNCIL REGULATION (EC) No 2406/96 of 26 November 1996 laying down common marketing standards for certain fishery products http://eurlex. 109-36. 328-9. Libbey L. 72-80. 4. and an Achromobacter species.R. 33.. 41. Do fish from tropical waters spoil less rapidly in ice than fish from temperate waters? Food Technology in Australia.M. Growth and activity of Shewanella putrefaciens isolated from spoiling fish. 1984. 2005. FAO Fisheries Report.A. Venkateswaran K. Ed. 38. 36. Processes in Pathology and Microbiology. physiology. Nealson K.do?uri=CONSLEG:1996R2406:20050602:E N:PDF Sumner J. Sumner J.E.M.J. Journal of Bacteriology. 1989.B. and regulation. Fish spoilage bacteria – problems and solutions. Iron and manganese in anaerobic respiration: environmental significance. Jørgensen B. Cohen D.H. 51-62. Applied Microbiology. Karnop G. Taussig M. 1982.. Occurrence and importance of proteolytic bacteria as spoilage indicators.H. 62-70. Dixon N. Brady P.. Food and Agricultural Organisation of the United Nations. Italian Journal of Food Science. Myers J. Satomi M. Dalgaard P. Magno-Orejana F. 35. Brochothrix thermosphacta. Dalgaard P.F. 71. 44. 48. Journal of Applied Bacteriology. Gorczyca E. 22 . Environmental Biotechnology. 40. 174. 42.. 262-6.. 1995. in Fresh Fish. 323-9. 32.H.FISH AND SEAFOOD 31. Huss H. Annual Review of Microbiology. 39. 1996. Kell D. Saffarini D.. 43. 33. 2002.L.. 9. Vogel B. 67.. Localisation of cytochromes to the outer membrane of anaerobically grown Shewanella putrefaciens. 6689-97. Gram L. 311-43. 61-6. 1984. 36. Oxford. The inhibition by CO2 of the growth and metabolism of microorganisms. 13. Lee J.. Blackwell Scientific Publications. a dominant microorganism in Mediterranean fresh fish (Sparus aurata) stored under modified atmosphere. Archiv für Lebensmittelhygiene.S. Do Tropical Fish Keep Longer in Ice Than Temperate Fish: The Circumstantial and Definitive Approaches. 1989. Drosinos E. Miller A.J.H. International Journal of Food Microbiology. Huss H.. Role of proteolytic bacteria in fish spoilage.. Applied and Environmental Microbiology. 1994. 334. Nychas G.. Quality and Quality Changes.eu/LexUriServ/LexUriServ. 3429-38... 317.M.R.europa... 1985. 34. 1973. II. Gram L. 18-21.
2000.L..M. Mac Rae I. 58. MacKinnon L. Whiting R.. Food Technology. Panicker G. Hammond R. Experimental evidence for toxin production by Aeromonas hydrophila and Aeromonas sobria in a meat extract at low temperatures. Slutsker L. Huss H. The International Commission on Microbiological Specifications for Foods. Bean N. PCR detection of a newly emerged pandemic Vibrio parahaemolyticus O3:K6 pathogen in pure cultures and seeded waters from the Gulf of Mexico.United States. Wilson S.J.and waterborne pathogens. USA. Journal of Infectious Diseases.L. 1661-6. Bej A. 181-8. Kluwer Academic/Plenum Publishers. 181.J. 47. Loessner M.C.H. in Microorganisms in Foods 6. The International Commission on Microbiological Specifications for Foods. Altekruse S.. Thompson S. Ray B. Majeed K. Ljungh A.. 55 (SS10). 1997. Microbial Ecology of Food Commodities... 48. 172-249.. Oxford.. Bishop R.K. Griffin P. 20. E and F in salmon fillets stored under modified atmospheres at low and abused temperatures. Embarek B. Painter J. International Journal of Food Microbiology.. detection and growth of Listeria monocytogenes in seafoods: a review. 303-12. Sampling for Microbiological Analysis: Principles and Specific Application.W..P. Applied and Environmental Microbiology. 1-34. 1986... Olley J. Morbidity and Mortality Weekly Report.. 1991. Embarek B. 100-3.M. Miller A. 181-96. Golden D. Daniels N. Myers M. International Journal of Food Microbiology. 12. International Journal of Food Microbiology. Genigeorgis C. 2000. 54. 12. 1973-1998.P. 55. 1993.. International Journal of Food Microbiology. 1987. 2194200. Jay J. Heat resistence of Listeria monocytogenes in vacuum packaged pasteurised fish fillets. 17-34. 53. Springer. Surveillance for Foodborne-Disease Outbreaks . 85-95. 2006..C. 52. Use of risk assessment to reduce Listeriosis incidence. 2003.. Presence. Lynch M.A.N. Lindroth S. 23 ... 46. Vibrio parahaemolyticus Infections in the United States. 1998-2002. Aeromonas and Plesiomonas as food. The International Commission on Microbiological Specifications for Foods..H. 50.A.. Garcia G. 56.. 57. 1-37. Fish and Fish Products (Microorganisms in Fish Products). Microorganisms in Foods 2. 69.. Woodruff R.K.K. 1994.CHILLED AND FROZEN RAW FISH 45. 51 (4). 330-6.M. 51. Risk of growth and toxin production by Clostridium botulinum non-proteolytic types B. Histamine fish poisoning revisited. International Journal of Food Microbiology. Lehane L. Journal of Food Protection. Ed.... Smith J. 2005. Wadström T. Modern Food Microbiology. Blackwell Scientific Publications. Braden C. 49. 23. 2005. New York. 1991. 50.
S.M. Olley J.. 51. Aberdeen.. Do Tropical Fish Keep Longer in Ice than Temperate Fish: the Circumstantial and Definitive Approaches.. Simard R. Torry Research Station of the Ministry of Agriculture. Cho Y. Houston N. 1992.9 1. Kitts D.. Akiba M.9. Prediction of shelf life and safety of minimally processed CAP/MAP chilled foods: a review.A. 741-50. Jones Y. The Microbiology of Seawater Fish. 62-70. 1984. International Journal of Food Microbiology.. Borgstrom G... 1989. Technomic Publishers. Ed.H. Detection of specific spoilage bacteria from fish stored at low (0 °C) and high (20 °C) temperatures. Rome. Magno-Orejana F. Eds.G. in Fish as Food.. Journal of Food Protection.C.M. 317 (Suppl. 1992. Torry Research Station. Food and Agriculture Organisation.1 Further Reading Processing Anon. 73-89. 13-32. Application of predictive microbiology to assure the quality and safety of fish and fish products.. Smith G.P. 15.9. International Journal of Food Microbiology. 6. 1. Huss H. Studies on the microbiological ecology of mackerel stored by the method of partial freezing.FISH AND SEAFOOD 1.2 Spoilage Shewan J.D. 317-21. in Torry Advisory Note No 93. in Seafood Science and Technology. Food and Agriculture Organisation of the United Nations. Further Studies on Marine Fish Stored Under Modified Atmosphere Packaging.. in FAO Fisheries Report No. Sumner J.. Huss H. Mathematical Modelling Used for Evaluation and Prediction of Microbial Fish Spoilage. Journal of Food Science. International Journal of Food Microbiology.. 1. Bulk storage of cod fillets and gutted cod (Gadus morhua) under carbon dioxide atmosphere. Changes in microflora and chemical compounds in mackerel stored by partial freezing.D. 1988. 1984. Ravn Jørgensen B.. Florida. 55. 1989. 1986. 1961. Gram L. 24 . 65-72. Shinano H..M. Cann D. 1997..... Ross T. Huss H. 4.H. Torry Research Station of the Ministry of Agriculture. Ogden I. 35. McMeekin T. Gibson D. Villemure G. Microbiological quality and shelf life prediction of chilled fish.H. Handling and processing of scad. Hobbs G. Dalgaard P. 295-307.E. Lancaster.1983. 1985. Trolle G. Ed.). 1. Bulletin of the Faculty of Fisheries (Hokkaido University). International Journal of Food Microbiology. Shahidi F. Picard G. Fisheries and Food.. Gibson D.L. Academic Press. Aberdeen. 127-34. Fisheries and Food.. Taoukis P. Labuza T. Estimation of the bacteriological quality of fish by automated conductance measurements. 271-85.
. Huss H. Otero A. 25 .. Sala F. London. Ed. International Journal of Food Microbiology.do?uri=CONSLEG:1996R2406: 20050602:EN:PDF Davies A. Applied and Environmental Microbiology.4 Standards and criteria Council Regulation (EC) Number 2406/96 of 26 November 1996 laying down common marketing standards for certain fishery products.H.L. Detection of specific spoilage bacteria from fish stored at low (0 °C) and high (20 °C) temperatures. 479-88. Inhibitory effect against pathogenic and spoilage bacteria of Pseudomonas strains isolated from spoiled and fresh fish. 1997. Eds. Amsterdam. Elsevier Science. 1992. in Fish processing technology.H. 1987.K. 1.. 59. Klausen N. 4.. 1. Garcia M...M. Gram L. Liston J. pre-incubation at low temperature and pH on the thermal resistance of Aeromonas hydrophila. Modified atmosphere packaging of fish and fish products. 72. 1993. 65-72. Trolle G.J. 5. 20023.europa. Journal of Applied Bacteriology. http://eur-lex. Hall G.H. International Journal of Food Microbiology. Blackie Academic & Professional. Huss H. Gram L.eu/LexUriServ/LexUriServ.9. A rapid method for detection of histamine producing bacteria. in Quality Assurance in the Fish Industry.. 137-46. 1987. 2197-203. Effect of culture age.CHILLED AND FROZEN RAW FISH Einarsson H.R. 322-6. Huss H.. Predicting the shelf life of cod (Gadus morhua) fillets stored in air and modified atmosphere at temperatures between -4 °C and +16 °C.3 Pathogens Condon S. 1992. Jacobsen M.9.
g. Few seafood products are sold ‘ready-to-eat’. 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.). smell etc. products of lesser quality. are used in chilled and frozen prepared meals since many of the attributes on which quality is assessed are not present or noticeable (e. or could be disguised by sauces. 27 . due to the early cessation of spoilage. The majority of seafood products produced require a cooking stage before consumption. it does not necessarily apply to the toxins that may have been produced. CHILLED AND FROZEN PREPARED FISH PRODUCTS Dr. 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.2. although recent developments in improved supply chains. This is no longer the case with many processed products now available in stores. The same used to be said for the difference between fresh and frozen products. Even though heating prior to consumption will eliminate pathogenic microbes. colour of gills.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. but still acceptable and meeting all safety criteria. emphasising the high quality of the seafood and the gourmet nature of the prepared products. colourings etc. Simon Derrick The Grimsby Institute Humber Seafood Institute Europarc Grimsby DN37 9TZ United Kingdom 2. clarity of eyes. smoked (hot) mackerel.g. the exceptions being Sushimi (high quality raw fish). seafood (with the exception of some crustaceans) remains uncooked until the point of consumption. Even products where the seafood is only one ingredient within the recipe (composite food products) it should be noted that unlike other meat-based products. smoked (cold) salmon. and crustacea such as shrimp and crabs. Sushimi tuna).
FISH AND SEAFOOD Recent trends in the market for food and seafood products. the processing industry has developed a wide range of value added products. Microbiological hazards include Salmonella. and these are increasingly being presented with an ever-expanding range of accompaniments. Marketing-style descriptions and process definitions are presented below to acquaint the reader with the variety and forms of seafood available. 2. Seafood is the term used descriptively to include all categories of product. from both wild capture and aquaculture sources. which can be farmed. The market spans fresh fish counter sales and frozen recipe dishes with fish as the predominant source of protein. Listeria spp. they are usually of a better standard than species such as tilapia. including freshwater fish. they are usually supplied filleted with head on. and this has produced a greater availability and diversity of products in the marketplace. but these are now finding favour as products in their own right. and are well known for their diet consisting of waste materials. or in MAP depends on catch conditions and treatment. They are typically flown into export markets. etc. molluscs. rather than frozen. and the consumer only has to heat (cook) the product.. Doubts have been raised over the microbiological status and handling practices of reef-caught fish. in particular. This includes all categories of seafood including trout and other freshwater species. and pathogenic Vibrio spp. 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. prawns. but should be equivalent to cold-water products.. have shown the increasing importance of convenience foods. Frozen fillets may have a surface layer of 28 . They can be with or without skin and may contain pin-bones although the major skeletal bones are removed during filleting. The expected shelf life attainable for this format is usually up to 8 days.2 Definitions The traditional focus of the chilled and frozen fish market has shifted over the last few years. Cold-water fish microbiological specifications are inappropriate for assessing freshness of warm-water fish. where the preparation has already been carried out. Shelf life on ice. At the same time. Wet or chilled/frozen whole fish are whole fresh fish that may or may not have been filleted. as they tend to lose colour and sheen after defrosting. shellfish and molluscs. Prepared natural fillets are fillets of fish without any additional coating. Nevertheless. It is sold from wet fish counters on ice or pre-packed in modified atmosphere packaging (MAP). Warm-water/reef-caught fish were initially supplied to add colour and variety to wet fish displays. There is also a limited market for filleted material. often using the simplicity of a microwave oven.
.2 days) longer than that of non-prepacked fish. VP products) because of the high salt content acting as a barrier to spoilage. in VP or through mail order. Fresh fillets are sold from wet fish counters. or pre-packed in MAP or vacuum packaging (VP). with a minimum residual salt content of 3. mackerel. natural fillets of fish such as herring. but may be increased (up to 1 month) by packaging methods like MAP or VP. and the product then shaped and heat-processed to cook the product before being chilled. swordfish steaks). The cold smoking process does not cook the fish or destroy microorganisms so the expected shelf life is not significantly increased from that expected for wet fish fillets. Fish steaks can be cut from a natural fillet or loin (e. the ingredients premixed. which is approximately 30% (or 1 . 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. salmon. and salmon. mackerel. Some products may be salted for flavour only (e. These products are packaged in separate high-care facilities to minimise contamination with microbes either from the workforce or from raw products. tuna steaks) or band-sawn from a frozen block (e. A core temperature of around 27 30 ºC for approximately 100 minutes is a typical processing condition. This process in effect cooks the product and reduces or eliminates microbial contaminants. 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.g. in addition.5%. The pâtés are sold from segregated ‘wet fish’ counters as a ready-to-eat product.2.5 .g. haddock. 29 . etc. A typical cold smoking process may pretreat the fillet by dipping in a 26% brine solution with a retention time of approximately 3 minutes. sawdust with other woods adjusted for flavour profile and burn duration).g. resulting in a product that is ready-to-eat without any further cooking. Again. a protective glaze may be added to the frozen steak. be flavoured with herb dips or peppercorns. Added colours can be included with the brine solution to meet specific requirements for products e. Hot-smoked fish are prepared from brined. The shelf life for MAP fillets is typically up to 5 days. They are sold from wet fish counters. Pâtés are prepared with raw fish. Cold-smoked natural fillets are prepared from a range of cold water fish including herring. They are then kilnsmoked to a minimum core temperature of 66 ºC for 2 minutes. or pre-packed frozen with added glaze.5% in the aqueous phase.CHILLED AND FROZEN PREPARED FISH PRODUCTS frozen water that forms a protective glaze. cod). The expected shelf life of these products is up to 8 days (or longer for some chilled. annatto with smoked haddock. Cold smoking is essentially a flavouring process.g. The result is fish with a typical residual salt content of 1. and typically uses natural smoke passing over the fillets suspended on racks in a kiln (oak chippings. which may.
fish nuggets/bites with dips and seafood-based snack products. jellied eels. etc. coated in batter or breadcrumbs. flavours. and may include molluscs or prawns that may or may not have undergone a minimal heat process during their primary processing. There is an increasing market for high value fresh or thawed fish (e. The cakes are usually coated using a three.g. e. It is often used as a protein supplement. Prawns.) Raw material is typically fish fillet. Scampi are tails of Dublin Bay prawn or langoustine. with an expected shelf life of around 7 days. Breaded fillets/steaks and battered fillets are either natural or shaped fillets. e. frozen blocks or canned fish and may include molluscs or prawns that may or may not have undergone a minimal heat process. or cut from a preformed frozen fish blocks into shapes. mixed whitefish.. 30 . Fish cakes are products produced by mixing fish with potato and herbs. binders. haddock. long-life seafood mixes using sorbate and benzoate as preservatives. taramasalata. and formed into round cake shapes. which may include molluscs. 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. and seafood mixes. addition of ingredients such as colours.g. Natural prawns from cold or warm waters can include those peeled. rice or pasta. ‘fingers’ before being coated in batter and/ or breadcrumbs.FISH AND SEAFOOD Seafood recipe meal centres (frozen or chilled) are places where these products are supplied without a source of carbohydrate. and may also include additional ingredients such as fresh herbs and spices. They are pasteurised products and treated as high risk raw materials. Other ingredients may be natural or reconstituted. and in the shell. cephalopods. Band-saw dust and fish trimmings from fishblock preparation materials may be used. rice.or a five-stage process. pasta or potato. This ‘catch-all’ category includes seafood salads. it can be used as a seafood analogue. Surimi is the insoluble protein fraction of fish after washing and processing. Other. The majority are products made from various types of whitefish (cod. With further processing. tuna.g. salmon) coated and sold as chilled products. and heat-fixed. The protein source can be fish fillet or blocks.
it may be used for other products or markets depending on the product specification. which may become a focus of contamination deep within the muscle. and his co-workers have described a large celled microorganism found in spoiling fish and identified it as Photobacterium phosphoreum (3). searing or bar marking). 6). 2). skin. For a summary of background information on the hazards associated with fisheries products and ingredients. and ingredients added to the final product. storage. will the saprophytic flora start to invade the surface flesh (1. as a result there is an increase in spoilage rate thereby reducing the product shelf life. fat content. and an increased risk from a potentially unsafe product (especially those that receive a minimal heat process. The Food and Drug Administration (FDA) has produced guidelines for handling and processing of tuna. In summary. sensory assessment of freshness etc. Contamination will. Recent public opinion has favoured the use of dolphin. The concern is that fish may be visibly acceptable but any subsequent washing process will have little effect on removing contaminants. other potentially pathogenic and spoilage bacterial contaminants may be introduced from a wide range of sources including the environment. including seasonal variation.and turtlefriendly long lining to the less discriminatory netting. and the potentially toxic by-products of histamine-producing bacteria (5. 31 . species (Bluefin>Bigeye>Yellowfin>Albacore). remain on the surface of the fish fillet except where “gaping” or separation of the muscle blocks occurs. and pathogenic flora associated with seafood. After capture and during any subsequent processing.e. or visible damage from gaff marks (4). on the whole.I. whilst gaping is also dependent on a range of other factors that effect the muscle composition. Frequent rebaiting and prompt removal of caught fish is essential to limit rapid spoilage. handling or preparation stage. The edible tissue of some species of fish are more susceptible to gaping than others.3 2. and have emphasised the origin and sustainability of the fish as key marketing points. and processing conditions such as dehydration or poor filleting practices. and intestines of the fish. i.CHILLED AND FROZEN PREPARED FISH PRODUCTS 2. Dalgaard et al. see Table 2. Tuna caught and discarded as unsuitable for sushi or other high-risk uses is often diverted for canning. is not used for sushi or other high risk (and high value) products. A primary role in fish spoilage has been assigned to this organism. product handlers.3. Grading is based on size of fish (>25 kg). Only after death. specifying internal temperature limits for fish and for limiting temperature abuse during handling and grading (1). the microbial flora of seafood is associated with the gills. Tuna that does not meet the highest of grades based on colour.1 Initial Microflora Seafood Other chapters have discussed in some detail the indigenous spoilage.
Aspergillus. Aeromonas. Enterobacteriaceae. yeasts Pathogenic microflora/toxins Salmonella. yeasts Lactobacilli. Staphylococcus. histamine Cryptosporidium.FISH AND SEAFOOD 2.I provides an overview of common raw materials. cereus Bacillus licheniformis. coliforms. and associated spoilage and pathogenic microorganisms. however. staphylococcal toxin. B. coryneforms. Clostridium perfringens. anaerobic spore-formers. Penicillium. toxin. Pseudomonas. Moulds Enterobacteriaceae Salmonella. Escherichia coli. shelf life determination.4. In addition. Clostridium botulinum. Fusarium.4 2. Staph. protozoa and viruses B. pseudomonads. 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. Bacillus subtilis B. product assembly. for chilled products. raw materials. Shigella. Bacillus cereus. and. moulds Bacillus Bacillus. moulds Bacillus. mushrooms. 32 .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. be considered at an early stage during new product development.3. cereus Carbohydrates Potato flake and powder Rice Pasta Flour Dried herbs and spices Dairy products including butter. moulds. nuts and fruit Associated microflora Lactobacilli. 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. cream. Listeria.1 Processing and its Effects on the Microflora Selection of raw materials/ingredients It is important that the potential microbial contaminants of all ingredients. and so are the potential microbial contaminants associated with them. intermediate processing. cereus Staphylococcus aureus. Table 2. Clostridium Lactobacilli. moulds Bacillus. TABLE 2. enteric viruses 2.2 Non-seafood ingredients The range of ingredients used in the preparation of chilled and frozen prepared seafood products is very diverse. spore-formers. by the HACCP team or by a hazard and risk assessment programme. streptococci. Vibrio cholerae. and milk Cheese Water/Ice Bacillus. cereus B.
to evenly raise the temperature of the entire frozen product from -20 °C or lower. 2. as a heat process is often required for functional or organoleptic reasons. line extensions and process reviews. Band-sawn materials or fish dice are normally promptly further processed.1 .4. All have the same intention. defined holding time at a chill temperature (0 – 3 °C). but it also poses a risk of cross-contaminating other products undergoing tempering.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. but it is assumed that any band sawing or dice process will introduce heat from blade friction. It is recommended that the principles of HACCP are applied to novel processes. Intermediate processing and use of chill for storage of work-in-progress will help to optimise available life of perishable components. and contamination from equipment to clean fish muscle. this may be an incidental effect. depending on the size of the fillet 33 . Figures 2.2 Raw material tempering Whenever frozen seafood is used as the raw material it is normally tempered before processing. The use of water for rapid thawing of product is not recommended since it not only rapidly thaws the product surface.4. Generic examples have been used to illustrate only the major points. there are a number of distinct methods used.5 illustrate basic principles in the preparation of chilled and frozen recipe dishes. and potential opportunities for growth of foodpoisoning bacteria.CHILLED AND FROZEN PREPARED FISH PRODUCTS Any preparation stage or process should be suited to the raw material. Effects on the microflora are minimal during tempering (7). Rapid controlled cooling of heat-processed components will help to minimise spoilage.2. 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. up to -5 °C so as to improve the cutting and handling properties during processing. the core temperature. but will also need to ensure that the optimum shelf life can be safely obtained. Where the raw materials are frozen.4. or the use of microwave technology. 2. as the full complexity of the processes used for specific products cannot be covered in a general text. Examples of tempering techniques include the use of heated air. For some components.3 Preparation of fish component of chilled and frozen recipe dishes 2. 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.3.
2. are placed into plastic boxes and covered in ice to preserve their condition.3.1 Whole-fillet / scampi Scampi (Nephrops norvegicus).2.Fish Cakes). On arrival at the processing plant. The scampi meat (tails) is removed intact from the shell using potable water whilst still frozen.2). Raw material preparation stages for chilled or frozen coated fish products 34 .1.Shaped Fillets.Scampi.1B . 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.1.3. after landing. the scampi is washed in potable water to remove foreign bodies.1C . by mincing and mixing with a binding agent (as described in section 2. drained and then refrozen for storage until required for further processing. Fig. quickfrozen. 2.1 (2. Individual Quick Frozen (IQF) fish fillets or steaks may also be coated using a similar process.1.7 days. Frozen cores are usually batter-enrobed. It is then size-graded. frozen in a nitrogen blast freezer and weighed into sacks for later processing. chilled or sold chilled on defrost with an expected shelf life of 6 . may not rise above 0 ºC during processing. Any broken pieces or fragments of frozen tail meat are kept for use in the production of a reformed scampi product.1A . The processing of three examples types are described in Figure 2.FISH AND SEAFOOD processed.4. 2.4. The “blown” meat is graded. with the coating heat-fixed.
This is especially the case with block-sawn dust. once the dry components have been rehydrated.off-cuts or belly flaps. off-cuts and by-products from other processes (maybe minced) are reformatted give a final product that is still 100% fish. Therefore. Herbs or seasonings used in the recipe may contribute additional spoilage organisms or. The mix is shaped and formed ready for packing or for enrobing. additional spoilage organisms may be introduced. quickfrozen. pathogens such as Salmonella and Listeria.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..g. from potato flake. material will need to be transferred swiftly to a chiller. Shapes may then be cut from the individual slabs using either hydraulic shape cutters or continued cutting with band-saw type blades (e. iii) Fresh/fillet/minced fish block. 35 . Waste material produced from this type of processing is often reused in the third category (fish cakes). or promptly processed to avoid further quality loss. ii) Fish-filleting waste .2 Shaped products This type of product falls into two categories.3. with an expected shelf life of 6 . or sold as a chilled product after thawing. or those where the trimmings.4. 2. 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. Fish cake cores may be batter-enrobed. chilled. that are to be reused for other foods. unless it is poorly handled. spoil more rapidly than the dry ingredients.4. The remaining dried ingredients are mixed with the fish. This type of material will be susceptible to contamination by Enterobacteriaceae and possibly Staphylococcus spp. but.CHILLED AND FROZEN PREPARED FISH PRODUCTS 2.1. Fish raw material can be obtained from a number of sources i) By-product of fish shaping or cutting/dicing process .1. are handled and stored as any other food product.block-sawn dust. with the coating heat-fixed. as the physical structure of the material has been destroyed. Spoilage of fillet pieces or trimmings will not be appreciably different from that of fillet. more seriously.3. It is for this reason that all by-products from primary processing. fish fingers). because of its nature. Waste fish generated as a by-product of another process will always be susceptible to spoilage and water loss. The fish component of the cake will. such as aerobic spore formers and Staphylococcus spp. those that use frozen blocks of whole fillets. and water is added to rehydrate the components.7 days.
g.g. Natural fillets d. is the addition of a pre-dust. Bacillus spp. A large variety of product formats and types can be batter-enrobed. a. Scampi tails are particularly susceptible to contamination with Enterobacteriaceae. The product then passes along a wire mesh belt through a ‘waterfall’ enrober that gives an even coating of batter to the entire product.FISH AND SEAFOOD In all of the above examples. which typically consists of flour. rusk or crumb. rapid automated processing with the minimum of delays will minimise product spoilage by pseudomonads. Tempered band-sawn materials b. with an expected shelf life of 6 . absorb any excess moisture on the surface of the fish portion. the crumb is poured onto the product followed by repeat enrobing and crumbing to complete the five-stage process. raising agents.e. such as Listeria and Vibrio spp. and possibly Staphylococcus spp. starch. quickfrozen. with the coating heat-fixed.4. If a crumb coating is required.7 days. Fish products.. flour. a characteristic flora will develop from the batter components. Spore-forming anaerobes such as C. Products are preformed and presented to the process (either three. 2. and if required. e. or sold as a chilled product after thawing. Control of temperature limits the potential for growth of spoilage organisms and pathogenic bacteria. spices.4 Battering/enrobing process Figure 2. fish cakes Portions or fillets may be batter-enrobed. vibrios and other fishassociated spoilage organisms. Initially. from flour. lactobacilli from the milk powder and flour. The function of this is to improve the adhesion of subsequent coatings. and Enterobacteriaceae from flour. i. IQF/pre-formed fillets c. e. rapidly multiply if strict temperature control was not maintained. as well as ensuring that there is minimal contamination from human processors. milk powder and spices. Breaded or battered fish fillets are fully heat-processed before consumption. some of this initial flora 36 . and to carry flavourings where required. chilled. which is product-dependent and not always required. which can occasionally be isolated from protein-handling processes. potentially. and may include milk powder.2 illustrates the typical stages of enrobing or breading. The first stage. 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. egg. perfringens could. any excess is removed with an air knife.or fivestage) frozen or chilled. Batter contains water. Over time. because of the intensive handling required during processing.
This can be achieved by frequent clean-downs and adequate hygiene. The origin of contamination for the two organisms is often unclear. dry batter mixes are all potential sources. cereus can proliferate under certain conditions. but because batter is heat-set.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. aureus and B. this is not of great significance to the microbiological standard of the finished product. replaced in part by process contaminants and bacterial flora from seafood. contaminants such as Enterobacteriacae can proliferate and develop during the process. and. but it is believed that poor hygiene regarding product and food handlers. Flow diagram outline of a typical three. It is advisable that some control mechanism is employed to prevent toxinforming organisms from multiplying to dangerous levels.2. time-limitation or production run temperature control. With inadequate temperature control of batter systems. The FDA has issued clear guidelines on acceptable 37 . Staph. in the case of B. and both of these can produce heat-stable toxins that can survive a batter heat-fix process. Fig. cereus.2.CHILLED AND FROZEN PREPARED FISH PRODUCTS associated with the ingredients will be diluted out.
3.5. Well-managed breading and pre-mix systems that are segregated from batter enrobers present little hazard (7). pasta or potato. Flow diagram outline for recipe dish ingredients preparation 2.3 illustrates vegetable processing and treatment for inclusion in chilled and frozen fish recipe dishes. For example.4. Figure 2. There are two main categories. The vegetables must withstand processing. rice.5 Recipe products These products involve the combining of seafood with other food ingredients as part of a recipe.FISH AND SEAFOOD temperature regimes and run times permitted for batter enrobers.2. to produce a specific product.g. canned fish may be used in their preparation owing to it having wider availability and low wastage.1 ºC for no more than 3 hours is recommended (1). Vegetable quality is an important characteristic often overlooked in the selection of vegetables for frozen and chilled products.4.1 Seafood recipe meal centres (frozen or chilled) These products can be defined as the central component of a meal. but they are supplied without a source of carbohydrate e. and must not show evidence of disease. Fig. The protein source can be fish fillet or block. 2. and may include molluscs or prawns that may or may not have undergone a minimal heat process during their primary processing. as this could introduce off-flavours and reduce shelf life. A temperature no greater than 10 ºC for 12 hours cumulatively or 21. 38 .
The presentation of vegetables has a significant effect on the product quality. i. control is more onerous. rice or pasta. However. Once vegetables have been blanched. The opportunity for growth of L. frozen block or canned fish. 2. they should be stored under temperature-controlled conditions. A number of examples of direct faecal contamination of vegetables exist in the literature (8).3 log reduction in bacterial numbers on the surface of vegetables.CHILLED AND FROZEN PREPARED FISH PRODUCTS The products will often be presented with vegetable toppings. even those with pH-modified dressings. encourage more rapid decay and loss of structural integrity. For example. For premium products. coryneforms.4. with cases of foodborne disease traced to Listeria monocytogenes. and may include molluscs or prawns that may or may not have undergone a minimal heat process. loss of quality. (7. but also as components in recipe meals. Washing with a biocide will at best achieve a 2 . the use of night soil or incorrectly applied manure or sewage sludge for ground preparation may contaminate produce with a wide range of pathogens. all-year-round fresh vegetables are preferred. and protected from any post-processing contamination. spore-formers. Blanching of vegetables can be an effective means of reducing or removing surface contamination.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. will disperse the saprophytic flora of vegetables . larger diced pieces allow for easier handling and a reduction in spoilage but are less attractive to the consumer. Although the processes are claimed to be superior to using unmodified hypochlorite for washing. Any assumptions regarding the application of good agricultural practice to produce should be made with caution. Salmonella typhi. with the accompanying release of additional nutrients by processing. any processing stage. monocytogenes should be taken into account when setting shelf life or holding times of vegetable components.lactobacilli. or breadcrumbs. colour and water will occur.5. chopping and mixing of material. This is a concern for chilled products that may have a shelf presence for up to 6 days including day of manufacture. coli O157. 39 . coliforms and pseudomonads throughout the mix. and may include sauce accompaniments. for example. and E. 8) will. Products must be washed thoroughly to remove pests and soil. and reduce spoilage organisms and potential pathogens. In an attempt to improve washing and reduce chlorine taint. and mixed whitefish). Raw material is typically fish fillet. The majority are products made from various types of whitefish (cod. pastry. Obvious signs of vegetable soft rot due to mechanical damage or infection by Erwinia spp. haddock. not just as accompaniments. acidification of hypochlorite and chlorine dioxide have been proposed and recommended as an improved alternative. treated as a highrisk ingredient. For example. if the process is not appropriately controlled. However.e.
6. where composite meals are heat-treated. or use barrier or hurdle principles to preclude the growth of pathogenic microorganisms. spore-formers can proliferate if adequate control of temperature is not maintained.1 Sauce manufacture The variety of ingredients and components used in sauce manufacture is extremely wide. Sauces can be multi-component preparations or more simple preparations. rely on the use of microbiologically sound ingredients. should be chilled to <5 °C as quickly as possible using either blast chillers or other equipment specifically designed for this purpose. This is a particular issue especially where products are heat treated to eliminate vegetative cells. B. with the range always expanding as new tastes and cuisines are developed. cereus are well known and are still a common issue in the UK (9). An incident involving Staph. such as butter garnish with black pepper. 40 . the preparation process must ensure the eradication of all vegetative pathogens from the ingredients. Products or dried ingredients contaminated with either spores of aerobic bacteria (e.4. cereus. Potato toppings made from flake or powder will normally have little associated spoilage flora. Placing such products in a standard chill store to cool down could result in condensation. can germinate and rapidly multiply. Alternatively. monocytogenes and other bacteria that may be present on chilled surfaces such as ceilings. if subjected to temperatures between 20 and 40°C. Spices and herbs added to form toppings along with breadcrumbs or other coating materials will similarly contribute a spore loading (primarily Bacillus spp. Appropriate handling facilities or equipment will be required to protect the product. and an increased risk of contamination from L. Chilled pasta is becoming more popular than dried pasta as a component in many product lines. and then cooled slowly allowing germination and re-growth.FISH AND SEAFOOD Food-poisoning incidents traced to rice and caused by B. Typically. especially in those products that have a vegetable component or rice accompaniments.4. Sauces or marinades are added to fish to broaden appeal or to accompany components. but temperature-abused pasta can also spoil rapidly by mould growth. aureus toxin is well documented (10.6 Preparation of non-fish components of chilled and frozen recipe dishes 2. On hydration.g. a heat process in excess of 73 ºC will be required to rehydrate this component fully.) to the product (12. Manufacturers of chilled products with an extended shelf life must recognise the potential for growth and spoilage of psychrotropic strains of B. a process equivalent to at least a 5D Listeria kill will be necessary. If the sauce contains modified starches for freeze/chill stability. such as high-risk/low-risk barriers and dispensing and/or holding at above 63 ºC. 13). 2. cereus). 11). For this reason all products.
flour products. that of a dairy-based sauce.CHILLED AND FROZEN PREPARED FISH PRODUCTS Fig. 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. Marinade-type products are becoming more popular. respectively.5 illustrate the recipe dish ingredients preparation and assembly and packaging. etc. 13). Acidified or low-pH sauce formats using acetic acid or vinegar normally require a lesser heat process for the equivalent effect. Without barriers. for instance.2. Flow diagram outline for recipe dish ingredients preparation Time limitation may minimise the potential for spoilage by process contaminants or at the dispensing unit. spices. suspended in an acidified sauce with an oil component.4 and 2. and handling of processed sauce can be less rigorous than. 41 . The format uses flavour compounds .4.herbs. 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. Figures 2. the sauce component will spoil rapidly and the possibility of vegetative pathogens from post-process contamination or raw components growing to significant numbers cannot be ruled out.
The oxygen-free conditions will however allow other anaerobic bacteria such as C.5.7. 42 . Increased levels of carbon dioxide will also result in a slight decrease in the pH of the surface tissues.7.1 Vacuum packing A range of fresh chilled products including salmon (smoked and fresh portions). generally carbon dioxide and nitrogen.2 Modified-atmosphere packaging Originally developed for the red meat industry. botulinum to grow.4.4. as it dissolves in the surface moisture to form a weak acid.7 Effects of packaging on chilled products Products with minimal additional ingredients may use packing techniques designed to minimise microbial growth.4. Flow diagram outline assembly and packaging of recipe dish 2. The objective of this method is to deny the aerobic bacteria (both spoilage and pathogens) the oxygen that they need to grow. unless the temperature is strictly controlled to below 5 °C throughout the supply chain.FISH AND SEAFOOD Fig. which may restrict either microbial growth or enzyme activity. the method replaces air in the sealed packaging with a defined mixture of gases. so replacing the oxygen necessary for aerobic growth of spoilage microorganisms. 2.2. and tuna loins are packed by placing the product in a plastic pouch and removing all the air. These include: 2.
5. This is considered to be the one major difference between fish and meat products. Not only will these organisms have a numerical advantage over other proposed spoilage organisms. Aeromonas spp. and therefore will have a greater opportunity to dominate other spoilage flora numerically over the course of product shelf life (15). but a conclusive case for pathogenicity has yet to be presented for Aeromonas spp. and finally flavour loss. the slow development of ammonical and sulphuric ‘off’ odours and flavours(16). Shewanella putrefaciens and Ph. Spoilage of this material is not expected to differ greatly from the processes described elsewhere.1 Chilled storage For chilled products or chilled components of frozen products. but they will also be readily adapted to the nutrient source and conditions. (20). The case for growth and contamination of products with Aeromonas is stronger. except for some crustacea. but little on in-pack spoilage. 43 . The literature has good information on spoilage of individual components of recipe meals.5 Spoilage During processing. At refrigeration temperatures (5 . at slightly higher temperatures (10 ºC). followed by moisture loss. Beyond this. phosphoreum. in numerical terms. but is not limited to. This is also true of chill storage and the distribution chain. where the protein component is nearly always heat-processed. Spoilage will be most readily detected in proteinaceous components of ready meals.10 ºC) Enterobacteriaceae are often found to proliferate and produce a characteristic type of spoilage that includes. advanced spoilage gives further changes in texture and the production of characteristic ‘off-odours’. However.. Nonetheless. temperature control is important in preserving chilled integrity of ingredients. the microflora of interest will be mainly psychrophilic or cold-adapted (14) spoilage organisms such as Pseudomonas spp. 2. is typically uncooked or at best lightly processed to retain eating characteristics. initially. It is not clear whether the additional contaminants from components or processing will accelerate this typical seafood-type spoilage. Vibrionaceae will succeed (17 ). have both been suggested as possible spoilage or pathogenic contaminants of chilled recipe meals (18). and Yersinia spp. and in influencing the final eating quality of the product. by a loss of texture. both organisms are capable of growth at chill temperatures. a large number of nutritionally non-fastidious spoilage organisms are commonly isolated from seafood. The fish component of recipe dishes. as this organism is a common contaminant of freshwater fish species (19).CHILLED AND FROZEN PREPARED FISH PRODUCTS 2.
44 .5. released by a pseudomonad with demonstrable activity to 0 ºC and a peak in production at 10 ºC (15).2 Frozen storage Maintenance of product temperature at <-18 ºC will not permit bacterial growth or enzymic spoilage... spoilage should not be considered a purely microbial phenomenon as taints and off-flavours can develop by non-microbial means (e. Products that are developed with an extended shelf life beyond the 6 . As a note of interest. 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.. where the variety of components and ingredients used in manufacture is varied and complex. Still. producing characteristic off-flavours. It is not unreasonable to expect that spoilage bacteria release many extracellular enzymes. 2. freezing also has a protective role in improving the safety of raw materials by. The type and nature of the enzymes and their activity will depend on the bacterial species.g.7 days typical of fish products/recipe meals must consider the potential for growth of non-proteolytic C. Pseudomonas spp. Bacillus spp. The conditions and freezing process most favourable for good-quality products will. the shelf life of products is limited by the natural spoilage process described earlier. and Aerobacter spp. enzymic processes are often ignored or overlooked as insignificant or unimportant because of the rapidity of fish spoilage.5. For example. Other examples of bacterial proteolytic and lipolytic enzymes produced by Alicaligenes spp. within dairy products are well known. Spoilage can be either a function of bacterial growth or the microbial release of free enzymes into solution. However. for example. however. Serratia spp.3 Enzymic spoilage Post-harvest changes in the quality of fish due to enzymic activity and rigor mortis are well known and understood (10). botulinum. also be those that have the least effect on the viability of microorganisms (21). auto-oxidation of fats/oils). a recent report examining fish gut bacteria from cold waters characterised a proteolytic enzyme. Characterisation of the enzymes has demonstrated residual activity even after a heat process that would destroy vegetative cells (24).FISH AND SEAFOOD 2. this should also take into account shelf life of product components used in the assembly of the final product. These have been proved to impart off-flavours. eliminating nematodes in products that do not have a heat treatment step. This also ensures that any post-process bacterial contamination or spore-forming bacteria in the product will not grow. which would warn the consumer of the potentially unsafe nature of the product. By retaining the intrinsic spoilage flora of seafood. This reinforces the need for prompt processing and temperature control of ingredients and finished products.
soil. Numerous quoted figures exist in the literature regarding cell counts required before toxin production occurs.e. Inappropriate holding times and temperatures will. A number of other Bacillus spp. may be of benefit within any risk assessment for chilled and frozen prepared fish products.2 Clostridium botulinum There are reports of rapid spoilage of vegetables by C.6. potato.. In addition. Counts of between 103 . 2. and Campylobacter are recorded in filter feeding molluscs such as mussels.3 Clostridium perfringens A study examining C. Vibrio spp. Hazards attributable to the non-seafood components are listed below and this information. with pathogenic microorganisms or viruses. permit rapid growth of B. cereus.104/ g should be treated with some concern (26). 2. Additional hazards from marine bio-toxins. perfringens in seafood noted a transient population of spores in the guts of fish feeding adjacent to rivers and sewage outfalls (29). etc. agricultural run-off or sewage can contaminate in-shore species. water. Cases quoting contamination of seafood with organisms such as S.6 Pathogens: Growth and Survival There are only two species of pathogenic bacteria that can truly be said to occur naturally in fish. especially molluscan filter feeders. Therefore.1 Bacillus cereus This is a common contaminant of dust. botulinum type E and Vibrio parahaemolyticus (25). 13. spices and herbs and vegetable shoots have been reported (12. along with previous discussion. low oxygen or anaerobic conditions. typhi. farmed or free-living crustacea (30). These are C. 2. and many raw or processed foods (26). 27).6. This may occur where storage under gas such as carbon dioxide is used to reduce ripening and so prolong storage. Germination of spores requires favourable conditions. and this issue may be of greater consequence for warmwater. air. perfringens contaminating seafood will depend on the source of raw material.6. botulinum when held in favourable conditions for growth (28) i. are dealt with elsewhere in this book. Incidents in such diverse materials as cream. any risk of C. have been linked with food poisoning and should also be considered. and some effects related to product type and time to toxin production have been noted. histamine intoxication. 45 . in some cases.CHILLED AND FROZEN PREPARED FISH PRODUCTS 2. or a heat shock followed by holding at a temperature suitable for growth.
Chilling may help Shigella to survive. monocytogenes as a contaminant in foods (31).5 Shigella spp. monocytogenes tends to grow poorly at refrigeration temperatures in seafood (33). 33) due to its ability to grow at low temperatures and form “biofilms” on surfaces. which is not affected by any subsequent cooking process. will colonise factory environments in a similar fashion except for possibly Vibrio spp. 2. Shigella spp.4 Listeria monocytogenes Interest in this organism has been characterised by a number of very serious incidents associated with a variety of product types (27). 2.6. growth may occur in foods (26). when introduced or inoculated into sterile components of chilled prepared recipe dishes.FISH AND SEAFOOD Vegetable raw materials are potentially contaminated through soil conditioning with animal manure. infected food handlers. It is unclear. the possibility of a sanitiser-resistant biofilm colonising processing environments should be viewed with some concern (32). indigenous or other. Rates of contamination with smoked seafood are much higher than those with unsmoked products. All the above points should be considered within any risk assessment of the potential for growth and toxin production by Staph. but under the appropriate conditions. These conditions will allow the production of the toxin. These include inadequate refrigeration and preparing foods far in advance of planned service. L. Sources of contamination are many because of the widespread distribution of this organism in the environment. the restrictions on growth rate seen with seafood will not apply. aureus (35).6. at the present time. monocytogenes has been frequently isolated from fish-processing plants and is thought to be able to colonise this type of environment (32. 46 . and this is believed to be due to a tolerance to brine used in the smoking process (32). L. 2. and will therefore cause “food poisoning” even in a product that is cooked before consumption. aureus enterotoxins in foods. An acceptance has now developed in the UK of the presence at low levels of L. However. if any other potentially pathogenic organisms. and inadequate cooking or holding within the temperature growth range (27). but.6.6 Staphylococcus aureus There are a number of underlying reasons for the development of Staph. 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).
The following tables (Table 2. Microbial guidelines. .II and 2. but provide the industry with an indication of the microbial level at which action is required to ensure product safety. 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. although the only seafood products specifically identified are raw “shelled and shucked products of cooked crustaceans and molluscan shellfish”.A dairy-based sauce that includes a non-pasteurised cheese will contribute significant aerobic total counts. In the EU these are defined under European Commission (EC) Regulation 2073/2005 (38). aureus enterotoxins and B. 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.CHILLED AND FROZEN PREPARED FISH PRODUCTS Illnesses caused by Staph.III based on IFST Guidelines (39)) provide example guidelines for seafood products. pastry. although due to long analysis times required. as published by for example. but these may be eliminated by a subsequent heat-treatment stage. and as such the monitoring is for verification purposes rather than control. the fresh product is often consumed before the results are obtained. pasta. such as batter mixes. with respect to the number and types of ingredients used.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.6. (36). and their association with similar materials. Microbial levels at the end of manufacturing (m) will be set much lower than those at the end of shelf life (M) which in turn should still be safe to consume.7 Published Microbiological Criteria Due to the complexity of many processed food products. are not mandatory. 2. etc. unless the criteria is for a pathogenic organisms where only non detection is acceptable. 2. Microbiological specifications do not necessarily relate to product safety. cereus emetic toxin are often confused owing to similar symptoms. and are a recognised means of providing data on manufacturing standards. For instance. It should be remembered that 47 . and not to final products. the Institute of Food Science and Technology (IFST) (39) or the British Retail Consortium (BRC) (40). Microbial criteria and the monitoring of microbial levels within a product are important for monitoring trends within production systems. it is not surprising that Microbiological Standards (defined as mandatory compliance required (37)) are limited to those raw materials in which specific hazards occur.
fish fingers.FISH AND SEAFOOD the microbial criteria for other products (rice. parahaemolyticus (warm-water fish) Histamine (scrombroid fish) Indicators and spoilage organisms Organism APC (heat treated) Enterobacteriaceae E.g. coli GMP* <104 <102 <10 Maximum Product-dependent 104 103 GMP* Not detected in 25 g Not detected in 25 g <102 <20 Not detected in 25 g <50 ppm (<5 mg/100 g) Maximum Not detected in 25 g 103 104 103 102 <50 ppm (<5 mg/100 g) *GMP = Values found immediately after production under good manufacturing conditions. monocytogenes B. coli GMP* <102 Maximum 104 TABLE 2. TABLE 2. pasta.II IFST Guidelines for part-cooked foods – e. 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. cereus Staph. aureus V. coated fish. batter etc) should also be consulted when establishing the specifications for raw materials and final product.fish products.III IFST Guidelines for processed foods . L. ready meals Storage: Frozen/chilled Use: Re-heated prior to consumption Pathogens and toxins Organism Salmonella spp. Maximum= Levels at end of stated shelf life 48 .
2. Outbreaks of foodborne illness in humans. Communicable Disease Report. Communicable Disease Report. Journal of Milk Food Technology. 59 (2). Powers E. International outbreak of staphylococcal food poisoning caused by contaminated lasagne.. 1311-7. 1991.L. Journal of Milk Food Technology.. 6. 15. Taylor S.. 8. 1976. 1997..K. 39 (10). Pathogenic microorganisms associated with fresh produce.. Food Science and Technology Today. Kluwer Academic.. 3.C.. 1962.C. Ohgiya S. 96. Letters in Applied Microbiology. Kumeta H. Hoshino T. 47.M. Toxin Related Diseases: Poisons Originating from Plants.. Latt T. 25 (1). Letters in Applied Microbiology. 233-7. London. 1997.R. International Commission on Microbiological Specifications for Foods.J. Sakamoto T.R. Journal of Hygiene. References US Food and Drug Administration. Ishizaki K.. 70-2. 167-93. Eds. 2001.. The bacteriology of fresh and spoiling fish and some related chemical changes. Beuchat L. Matsuyama H. Animals and Spoilage.. Collette R. Wienke A. Brown T... Bacterial histamine production as a function of temperature and time of incubation.L.C. Behling A. 5 (3). 1997..M. Microorganisms in Foods 6: Microbial Ecology of Food Commodities. Washington D.A. 67-73. Microbiology of Marine Food Products. Martin R. 9. Aureli P. in Fish Inspection Quality Control and HACCP: A Global Focus. 12.E.L.H.R. Isolation of a Pseudomonas species from fish intestine that produces a protease active at low temperature. 166-73. Mejholm O. Powers E. . Dalgaard P... Masuoka Y. 24 (5).. Cambridge. 373-8. 14. Food and Drug Administration. 663-87. Recent Advances in Food Science. 13. 1976.R. Journal of Food Science.8 1. 1986. Hobbs G. England and Wales Quarterly Report. Ward D. IBH Publishing Co. Woolaway M. Lawyer R. Fish and Fisheries Products: Hazards and Controls Guidance. Journal of Food Protection. 11. Microbiology of processed spices. 5. Importance of Photobacterium phosphoreum in relation to spoilage. Bartram P.M.W. Bartlett C. 1. 38 (11). New York. Van Nostrand Reinhold.. Yumoto I. Proper processing and handling of raw tuna. Slavin J. 2005. Oxford.. 215.CHILLED AND FROZEN PREPARED FISH PRODUCTS An authoritative survey of specifications can be found in Shapton & Shapton (41). Fish: microbiological spoilage and safety. Taylor and Francis. 1982.. Christiansen T. Incidence and levels of Bacillus cereus in processed spices. 10. 2.R. 1997.. Hackney C.T. 7 (24).G. 668 -70. 204-16. London. 1993. 49 4. Huss H. Pvt. 7.. 1991.J. Gilbert R. Murrel H. Ltd. Shewan J. Tu A.
cereus and C. 28. Baird-Parker T. Anon. Her Majesty’s Stationery Office. Ronner U. 1990. Blackwell Scientific Publications.L. perfringens? International Journal of Food Microbiology.. International Commission on Microbiological Specifications for Foods. Microbiology in fishery science.. Eds. Barnes E. Gould G. Leistner L. 2000. Halligan A.C. 4. London. Detection of specific spoilage bacteria from fish stored at low (0 ºC) and high (20 ºC) temperatures. Leatherhead. What problems does the food industry have with spore forming pathogens B. Wood L. .J. Jemmi T. Survey of the incidence of Aeromonas and Yersinia species in retail foods..... 21. Fishing News Books Ltd.J. Ray B. 472-506... 1994. Leatherhead Food Publications. Lund B. in Anaerobic Bacteria in Habitats Other than Man. 40 (5). 1995. 11. 4 (1).C. 51-158 Saxby M. Trolle G. 1979.V. Morgan D.H. 13. 2005. 1988. Lund B. Blackie Academic Press. 333-6. Aeromonas and Verotoxigenic E. 20.A. Matches J. Occurrence of Listeria monocytogenes in fresh water fish farms and fish smoking plants. 32. Applied Microbiology. 31. 65.. Liston J. Fresh and processed fish and shellfish . Effects of freezing and storage on microorganisms in frozen foods: a review. Walker S. Journal of Food Safety.FISH AND SEAFOOD 16. Is Aeromonas spp. U. International Journal of Food Microbiology. Jay M. Aspen Publishers. Advisory Committee on the Microbiological Safety of Foods. Blackie Academic and Professional. Modern Food Microbiology.M.M. SAB Symposium Series No. Brooks J. 29. Springer. 351-72.W. Eds. New York. 59-72. Food preservation by combined methods. in Advances in Fish Science and Technology. PHLS Microbiology Digest. Granum P. 34-40. 27. 13 (1). Food Microbiology.R. 26. 18.. 22. Mead G. 1992. Liston J. 655-60. Food Control. coli (VTEC) . 1977. Food Research International. 1996. Ed. Andersson A. 1974. 30. 274-86. Connel J. 28 (4). 1992.H. 25 (2). Glasgow. 50 17. Speck M.R.Yersinia. London. 24. Clostridium perfringens in the environment. Curran D. Gaithersburg.J. Food Taints and Off-flavours. Microorganisms in Foods 5: Microbiological Specifications of Food Pathogens.. Anaerobes in relation to foods of plant origin. Gram L.. 19. 1986. Oxford.. 41-3. in The Microbiological Safety and Quality of Food. Report on Vacuum Packing and Associated Processes. 1996. The Emerging pathogens .. 309-16. a foodborne pathogen? Review of the clinical data. 1987.E. Huss H. 9. Journal of Food Protection. 23. 1993.. 155-9. Golden D. Gram L.J. 1996. Microbiological guidelines for some ready-to-eat foods sampled at the point of sale: an expert opinion from the Public Health Laboratory Service.. Keusch A.C. Huss H. 25. Loessner M.A Literature Survey.K.J. 28.
Shigella as a foodborne pathogen. 36.co.do?uri=CELEX:02005R2073-20071227:EN:NOT Institute of Food Science and Technology. Cambridge. Varnam A. 1997.H. 2006. 37. http://www. 51 . Development and Use of Microbiological Criteria for Foods. British Retail Consortium. Guidance on the Practical Implementation of EC Regulations on Microbiological Criteria for Foodstuffs.campden.L. 34. 38.A. 35. 788-801. CRC Press. http://www. 60 (4). Evans M. Chilled Food Association.F. Chilled Food Association. U. Institute of Food Science and Technology.org/Resources/Chilled%20Food%20Association/Public%20 Resources/BRC_CFA_Micro_Criteria_Guidance_Ed_1.. 1987. Smith J.CHILLED AND FROZEN PREPARED FISH PRODUCTS 33. Journal of Food Protection. 40.eu/LexUriServ/ LexUriServ.pdf Shapton D.chilledfood.pdf Commission Regulation (EC) No 2073/2005 of 15 November 2005 on Microbiological Criteria for Foodstuffs. Psychrotrophic Bacteria in Foods: Disease and Spoilage. London.2. Principles and Practices for the Safe Processing of Foods. 1991. Phil Voysey.uk/cday/posters2008/npd6. London.G. McCarthy S. Shapton N. Journal of Food Protection. 41. Gloucestershire. Wolfe Publishing. Woodhead Publishing Ltd. 39.K. 372-6. 1992.europa. Kraft A. Incidence and survival of Listeria monocytogenes in ready-to-eat seafood products. Microbiological Criteria.K. 50 (9). Foodborne Pathogens an Illustrated Text. 1999..A. U. 1998.A. Campden & Chorleywood Food Research Association. http://eur-lex.
cockles. oysters. Food and Drug Administration 22201 23rd Dr. the shell. squid. In the process of filtering water.S. Gastropoda. slugs. cockles and scallops. Jr. P. snails. Pacific Regional Laboratory Northwest U. oysters and clams can process about 3. while 53 . where it is eventually swallowed and digested. and shrimp. Bivalves feed by filtering large volumes of water across their gills to obtain oxygen and food. mussels.6 litres. Plankton and algae form the bulk of their diet from seawater. O. lying well within the mantle cavity. chemical contaminants and other impurities on the mucus of the gills. lobster. octopi. MOLLUSCAN SHELLFISH Carlos Abeyta. abalones.7 litres of water per hour. and scallops. clams. The term shellfish in this context is limited to oysters. Other species of shellfish such as crabs. S.3.1 3. and Cephalopoda. which includes oysters. viruses. cockles. For example. completely encloses the body. The class having species of economic and public health importance is the Pelecypoda. can hold the shells tightly shut against enemies (1). Pelecypoda. Pelecypods have little need for locomotion. scallops. the adductors.E. composed of two valves hinged together dorsally. Box 3012 Bothell WA 98041-4421 United States of America 3. The digestion process requires less than 2 hours in actively feeding adult bivalves (2). respectively. mussels. Strong muscles. Scaphopoda.1 Definitions General features of the mollusc The phylum Mollusca.1. The phylum consists of five classes: Amphineura. Many microorganisms ingested by shellfish survive the digestive process (3. and tooth shells are a group of soft-bodied animals that usually secrete external protective shells. The class Pelecypoda includes all of the bivalves such as clams. 4). Special tracts of cilia move this mucus toward the mouth. the foot is compressed to form a muscular spade for digging and the head is greatly reduced. mussels. clams. In these forms. they also trap bacteria. chitons. whelks. and 2. In most species.
and usually at higher concentrations than found in their habitat. The following are descriptions of the various bivalves and their forms of preparation for human consumption (6): TABLE 3.North America Saxidomus nuttali Venus mortoni Protothaca thaca Meretrix spp. Mactra sachalinensis Saxidomus giganteus Japan Pacific – North American Saxidomus nuttali Protothaca staminea or Paphia staminea 54 .I).2 International commercial products Globally.1. many species of bivalves are edible (Table 3. 3. Clams are benthic and live burrowed in the mud. 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.I Partial list of bivalve molluscan shellfish products in commercial use internationally (5) Common name Clams Soft (shell) clam Pacific/Atlantic North America Pacific/Atlantic North America Pacific . The habitat of oysters and mussels are sessile.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.FISH AND SEAFOOD commercially valuable. dwelling on the bottom or attached to structures in the water column. but maintain contact with the water by means of a siphon tube (5). 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.
Different species are variously called quahog. or siphons. hard-shell or soft-shell clam. A mantle tissue encloses the body of the clam.2. Buried clams leave tubes. laterally compressed. and other names. Most clams are only a few centimetres in their maximum dimensions. extended above the surface to maintain water currents needed for respiration and feeding. 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. geoduck. mostly marine bivalve molluscs. but 56 . littleneck.1 Clam Clam is the common name of a large group of often edible. cherrystone. and two symmetrical shells (valves) held closed by two large muscles and joined by a dorsal hinge joint.FISH AND SEAFOOD Mussel Mussel Mussel Scallops Scallop Coquille St. The muscular foot is used to burrow in mud or sand.1.
either whole or minced. tidal waters or in brackish water in the mouths of rivers. uncooked (shucked). hermetically sealed. Fresh: in shell. 57 . Canned: clams are steamed open and the meats removed from the shells. smoked meats are also canned. having rough and irregularly shaped shells that are closed by a single adductor muscle. 3. 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.2 Oyster Oysters are marine bivalve molluscs. uncooked meats packed unprocessed. boiled and then sun dried for 3 to 10 days before packing in boxes (Hong-Kong) Smoked: meats usually canned in edible oil. Live: in shells. Canned soups include clam chowder. then heat-processed.2. and no siphon. uncooked. Smoked: meats. sun-dried. Frozen: in shell. These molluscs feed on minute plants and animals carried to them by the current. clam madrilène and clam liquor. Dried: shelled meats.1. after precooking and brining. in cans with either brine or clam liquor. trimmed and packed. Oysters lack a foot or have a reduced foot. oyster soup. washed. Frozen: shelled meats. or shelled meats. all in cans. Live: in shell Fresh: in shell or shelled meats. or shelled meats. Dried: Hoshigai (Japan) shelled meat. skewered with bamboo sticks. packed in weak brine. Soups: oyster stew. the meats are smoked and packed in oil in cans or jars. Canned: meats removed from the shell by steaming. They live free on the bottom or adhere to stones or other objects in shallow water along the seacoasts. oyster bisque.
are united. cooked meats packed in dry salt for transport. with the posterior ends of the shells (valves) broad and rounded. Fresh: shelled meats (often only the adductor muscle and the roe are eaten. brine. in which the individual branches. 58 . vinegar solution. scallops can swim or leap by a series of rapid closings and openings of the valves. The familiar marine mussel is asymmetrical. They have filibranch gills. Canned: cooked meats.2. Semi-preserves: cooked meats with vinegar-acidified brine and spices. In some species. or filaments.1. boiled. They have a radially ribbed shell with the edge undulated and swim by opening and closing the valves.4 Scallop Scallops are marine bivalve molluscs living in shallow waters of protected bays with sandy or muddy bottoms. in mussel liquor. smouldered and afterwards dried (Japan). sauces. The common scallop is about 5 cm long. mussel in butter sauce. composed of striated muscle fibres (used for fast swimming) and unstriated muscle fibres (used to hold together the two halves of the valves). and held apart.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 posterior halves of the valves contain a large adductor muscle that holds the valves closed. gutted. Fresh: cooked meats. Salted: cooked meats bottled in brine. The shells are fluted and round with a broad. Bait: used live extensively for baiting fish hooks for line fishing. The adductor muscle is the only edible part of the animal. also with other ingredients. Smoked: meats may be canned in edible oil. Unlike other sessile bivalve molluscs. the shell is often characterised with a lustrous nacreous or pearly lining inside their shells.1. 3. or packed with jelly.) Dried: peeled. flattened ear or wing at the hinge area.FISH AND SEAFOOD 3. and the anterior end smaller and pointed.2. Internally. scallops possess a single large adductor muscle. Live: whole with shells. mussel paste. by interlocking tufts of hair-like cilia.
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. Raw and partially cooked shellfish.2. Whatever contaminants are in the water will eventually get into the shellfish (7). nutrient concentration and pollutants of the water column. Because molluscan shellfish are filter feeders.1. The microflora are dependent upon a number of factors such as season. and butter. they accumulate pathogenic microorganisms from polluted waters. Fresh: meats removed from the shell by boiling. in sauce. 3. 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 . clams and mussels. depending on length of journey. The initial microflora of molluscan shellfish is comprised of the natural commensal microorganisms and the microorganisms accumulated from the water during feeding. also bottled in brine.MOLLUSCAN SHELLFISH Frozen: shelled meats. Vinegar cured: bottled in malt vinegar after brining. Primarily inhabits shallow waters near tidal zones where they bury themselves in the sandy or muddy bottoms. Salted: meats either lightly or heavily dry salted. and the environment. salinity. Cockles range from about 1 to 15 cm in length and have two valves of equal size. which affects the temperature. mostly in oysters. 3.5 Cockle A marine bivalve mollusc having a shell with convex radially ribbed valves. Canned: in brine. are vectors of disease transmission (8). Canned: in own juice. pH.2 Initial Microflora The microflora of molluscan shellfish directly reflects the environment from which the shellfish is harvested. Live: in shell.
The largest groups found are the Pseudomonads.1 Processing and its Effects on the Microflora Oysters..000 /g. They are termed viable non-culturable microorganisms.3..2. 3. whole or in part.. Water temperatures affect the concentrations of microflora in the water. and Aeromonads. Bacillus spp. dredges. 15). The microflora of molluscan shellfish at harvest consists predominantly of Gramnegative rods while the Gram-positives constitute a minor portion of the flora. the initial flora composition and concentration of bivalves are highly variable. have been observed microscopically in the digestive tract of Eastern and Pacific oysters. non-B Snow Mountain virus Plesiomonas shigelloides Salmonella spp. and shucked shellfish that is shellfish. and cockles The processing of bivalves takes two forms. In summary. Flow diagrams of the major processing steps of molluscs are presented in Figures 3. Vibrios. nutrients.1 and 3. Alcaligenes spp. Temperature. mussels. Moraxella spp. clams. Pseudomonas spp. For example. 3. salinity. 10. 11. molluscan shellfish have an Aerobic Plate Count (APC) of approximately 1000 to 100. There are microorganisms.3 3.1. Campylobacter coli Bacillus cereus Hepatitis non-A.3. Campylobacter jejuni Escherichia coli Hepatitis A Norwalk virus Other viral agents A variety of commensal microflora are found in molluscan shellfish. and yeast cells of Rhodotorula rubra and Trichosporon spp.3.FISH AND SEAFOOD Aeromonas hydrophila group Staphylococcus aureus Shigella spp.1 Processing description The method of processing shellstock begins with the harvester collecting shellstock by the utilisation of tongs. spirochetes of the genus Cristispira and Saprospira. but have never been cultured (14.1. observed microscopically. Flavobacterium spp.1.1 Shellstock 3. rakes. Micrococcus spp. (9. mechanical harvesters.. At harvest.. 12. Acinetobacter spp. 13).. Enterococci spp. Other microorganisms found in lesser numbers are the Achromobacter spp. shellstock (live) which is shellfish in the shell. from which one or both shells have been removed. pollution point sources and hydrological events will greatly influence the initial microflora in bivalves. responding to the environment in which they are harvested.. or by 60 . that have never been cultured on laboratory media...
by truck or boat.MOLLUSCAN SHELLFISH hand (Figure 3. Generic flow diagram of live shellfish from harvester to point of sale 61 . Shellstock are given a wash to reasonably free them of bottom sediments and detritus as soon as practicable. 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. The shellstock is transported dry. Secondly. Shellstock are placed in drycooler storage containers in the facility. or from other sources approved by the State Shellfish Control Agency (SSCA). for transportation to the point of sale. or immediately placed in refrigerated trucks. Thirdly. wet storage may be used to increase palatability by increasing salt content of shellfish harvested from low salinity waters. after harvesting. Fig.1. shellfish can be harvested from remote areas at convenient times and held live for brief periods at locations close to the point of sale. There are three primary reasons for wet storage: first. usually the boat will be docked by the processing facility with shellstock piled on the deck of the boat or dock. Shellstock may be wet-stored at the processing facility in tanks of clean seawater. Water used for shellstock washing is from approved growing areas.1). in containers from which they can be retrieved easily. If by boat.3. or in bay-approved shellfish-growing waters prior to sale or processing. usually in open-mesh containers and/or burlap sacks.
Food and Drug Administration (FDA) recommends that potentially hazardous food such as shellfish be held at 7. without adequate refrigeration. At harvest. Several studies have established that pathogenic Vibrios and other bacteria may be present in marine sediments throughout the year (21.1. and thus contaminating the shucked meat at the point of sale. 16. and Citrobacter species) and Vibrios. Excessive sediment on the shells and dead shellfish may increase bacterial 62 .1. 26). Shellstock should be refrigerated at temperatures at or below 10 ºC (50 ºF). when it reaches the processing facility. coli.FISH AND SEAFOOD 3. hydrophila (18). metabolic waste accumulates in the shell serving as nutrients for bacterial growth. 21. Washing shellstock helps prevent quantities of sediment and bacteria being mixed with. synthetic seawater for the purpose of de-sanding and salt uptake. At this temperature the multiplication of faecal coliforms (E. the bacterial counts in the meats will increase. Oysters will remain alive in the shell for approximately two weeks if kept at the recommended temperatures. Enterobacter. Wet storage operations are highly variable and may range from temporary storage near-shore in approved areas to on-shore tanks using re-circulating. however. shellstock are placed in refrigerated trucks for transport or placed in wet storage. for a considerable length of time. with the exception of A. The most important factor in controlling bacterial growth in shellstock is temperature. Shellfish in wet storage tanks are similarly subjected to pollution if the tank water is obtained from a polluted source. 23. Klebsiella. involving 691 cases was attributed to oysters contaminated in a wet storage area (27). The safety of shellfish is predicated by the cleanliness of the growing area waters from which they are harvested. with prolonged storage the microbiological quality of the bivalve decreases. to avoid possible contamination of the shellstock.2 ºC (45 ºF) or below (19). are prevented. 17). Harvest conditions are another factor that influences the growth of bacteria in shellstock. handling.2 Effects of microflora during processing The initial microflora found on or in shellstock is influenced by sources of shellstock. Generally. Shellfish may contain higher levels of pathogens than are found in the water in which they grow. and temperature storage of shellstock to point of sale. There is a positive relationship between sewage-polluted shellfish and enteric disease transmitted by shellfish (8. some pathogenic organisms will survive and multiply in shellfish (20. Over a period. Several studies have established the increase of bacterial counts in shellstock during post harvest period before the shucking process (18. An outbreak of infectious hepatitis in Sweden in 1956. and the sanitary practices applied during harvesting and shipping. 25). 22). Water used for shellstock washing should be of good sanitary quality. bivalves tightly close their shells thus trapping water and microflora. there is evidence that in the absence of adequate refrigeration. 24.3. Furthermore. including V. The washing of sediment and detritus soon after harvesting will prevent further contamination. If long delays occur before reaching the processing facility and/or refrigeration. Generally. vulnificus. shellstock are not refrigerated on the harvest boat.
1. In addition. If shellfish are not reasonably clean at the time of processing. Thus. through which the washed meats are discharged for rinsing and then packed in containers. 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). for the effects of microflora during the processing of shellstock. Due to the nature of the shucking operation. a considerable quantity of the adhering material will be mixed into the shellfish during the shucking process. 3. lobsters.1 Processing description The processing of shucked shellfish is limited to washing of shellstock.3.3.MOLLUSCAN SHELLFISH loads in the tanks and lead to increased microbial levels in the shellfish during storage. the tank is opened through a drain gate and chute. In the blower-washer. After washing of meats. clothing becomes very soiled. the knife is used to cut the remaining adductor thus freeing the shellfish (meats). Studies have shown the use of rubber gloves reduces the contamination level. thus contributing to high bacterial counts in the final product (28). Sealed containers are placed in boxes and covered with crushed ice for chilling or refrigeration and transportation. whereas cotton become wet and 63 . the finished product may become contaminated.2.1. Shucking and washing of meats under sanitary conditions prevents the proliferation of bacterial populations. Containers are placed in packing boxes for shipping at refrigerated temperatures.2 Effects of microflora during processing The same considerations as previously discussed. are applied to the shucking operation of shellfish. removal of shellfish (shucking). Another potential risk with the use of wet storage is the commingling of bivalve molluscs with other marine species such as crab. washing. Washing of meats with potable water is accomplished by placing the meats on a skimmer table (the stand-supported.3. To remove the shellfish attached to the remaining valve. the meats are in a tank of water and agitated by the introduction of air from the bottom of the tank. and packing shellfish into containers (Figure 3. 3.2 Shucked shellfish 3. Shuckers routinely use either cotton or rubber gloves during the shucking operation. Meats are collected in either perforated or non-perforated buckets for delivery to the washing station. and fish. 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.2).2. Under these conditions if the shuckers enter the packing room.1. shellstock is subjected to contamination when stored on the floor in standing water. It presents a risk of cross-contamination from the non-molluscan animals to the bivalves. or subjected to splash from foot traffic. washing and culling shellfish prior to storage is essential.
aprons. and gloves help prevent the contamination of shellfish. within one hour. There is a tendency to store such articles on the shucking benches or in packing rooms. Shucking buckets and storage containers are kept at adequate height above the floor to prevent contamination from floor splash. 2. Separate rooms or lockers for storage of clothing.2 ºC or less. and sanitised before each filling. Shucking buckets are completely emptied at the packing room and no overage is returned to the shucker. 4. Shucked meats are thoroughly drained.2. cleaned and delivered to the packing room or prechilled and placed in temporary refrigeration (≤ 2 hours) at 7. 3. 64 . 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.FISH AND SEAFOOD retains bacteria during the shucking thus increasing the bacterial load in shellfish (29). Fig 3. which may lead to contamination of product. Shucking containers are rinsed clean with running water.
Pseudomonas.47 to 6. 34. keeping shellfish refrigerated is the most effective method of retarding deterioration and spoilage. are an excellent medium for growth of bacteria. and packing the shellfish in a timely manner. streptococci. Earlier studies.2 ºC or less. skimmers. Flavobacterium. Refrigeration and shipping of shucked shellfish in wet ice is highly recommended. spoilage will occur. whereas bacterium such as Pseudomonas. rough. Shellfish. and proteolysis (10). Shucked shellfish are an excellent medium for the growth of bacteria. 35. Controls that should be applied include using clean packing equipment. 36). 3. 33. These organisms may play a significant role in the spoilage processes. these include lactobacilli. In summary. Micrococcus. 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. thus producing acid and gas (38). Achromobacter and Flavobacterium levels decrease during storage. particularly shucked meats. Lactobacilli seem to be the predominant microflora in shellfish.2 ºC. The spoilage process in shellfish may be divided into three stages: increase of acidity. is stored in the bivalve as a source of energy. shucking pails. For example.4 Spoilage There is a lack of information in the literature regarding spoilage microorganisms in shellfish. and that storage in wet ice is the most effective method for refrigeration of shucked meats (30. Temperature abuse will result in high bacterial counts. glycogen levels range from 0. blowers and other utensils that have cracked. Proteus. Shucked shellfish are held and transported at temperatures of 7. This process changes the bacterial profile to microorganisms favouring low pH. and yeasts. and Pseudomonas fluorescens are the responsible agents of spoilage in oysters held at various temperatures (10. Much of the work has been done with oysters and clams. Alcaligenes. The initial flora found in shellfish is the major factor of spoilage in bivalves. anywhere during the process of shellfish from harvesting to the point of sale. 31. Glycogen.8% (37). particularly oysters stored at 7 ºC. All equipment coming in contact with the shellfish such as colanders.MOLLUSCAN SHELLFISH Unacceptable practices in the packing step can affect the microflora of shellfish. Glycogen is hydrolysed by the initial flora found in shellfish. As described previously. showed that organisms such as Achromobacter. The environment has an effect on the bacterial population found in shellfish. 65 . a carbohydrate. in oysters. abundant gas production. or inaccessible surfaces are apt to harbour accumulations of organic material in which bacteria may grow. Studies have shown that bacterial growth is significantly reduced at storage temperatures of less than 7. 32). clean gloves. at the turn of the century. where there is a breakdown of temperature control or prolonged storage of shellfish.
The control of growth and survival of these organisms can be reduced by the use of good manufacturing practices including immediate and proper refrigeration.5 cases/100.1 Vibrio parahaemolyticus V. 66 . Major outbreaks are associated with the warmer months. Diseases caused by Vibrios are enteric in nature. widespread in both polluted and unpolluted waters. there were over 250 cases of illness on the West Coast of North American extending from California to Canada. and harvesting of shellfish in the cooler months. V. parahaemolyticus possibly producing illnesses (40). The majority of the illnesses were associated with the consumption of raw or partially cooked oysters and clams. where outbreaks occur regularly. approximately 23 cfu/g. parahaemolyticus causes gastroenteritis lasting 24 . parahaemolyticus multiplies rapidly at temperatures above 20 ºC. This bacterium is present in all the major processing steps of a shellfish plant. parahaemolyticus. and very common in Japan. headache and fever.5. Between May and September 1997.e. The ability of the bacterium to cause gastroenteritis is highly correlated with the production of a heat-stable haemolysin (42). ranging from epidemic cholera to sporadic cases of diarrhoea.FISH AND SEAFOOD 3. 41). The incidence of V. and to the point of sale. only a small percentage of strains in the marine environment are potentially pathogenic (45). They can be introduced to the shellfish from the environment or added during processing. One of 250 cases reported involved shucked products with low levels. of V. although non-hemolytic strains have been associated with illness (43. parahaemolyticus from shellfish is low. Fortunately. there is a recent report of low levels of V.1 Vibrios Vibrios are Gram-negative facultative rods naturally occurring in the marine water (estuaries.1. nausea.5 Pathogens: Growth and Survival This section deals with bacterial pathogens frequently present in shellfish. 3. 3. V. prevention of cross-contamination. coastal areas). diarrhoea. These organisms can be present in the shellfish from the point of harvesting. parahaemolyticus illness is usually a result of eating raw or partial cooked shellfish (8. during processing. primarily in brackish or saltwater. from harvesting to finished product (i.48 hours with abdominal pain.5. however. It is a common marine bacterium. lot and retail outlet. Descriptions of outbreaks or reports of growth of the organism in shellfish are presented. 44). and estimates based from a hospital study show less than 0.000 population/year (46). Large numbers of bacteria must be present to reach potentially infective levels of >105 colony forming units per g (39). Products involved were from the same processor. shellstock and packing of shellfish in jars) (29). Sporadic cases are frequent along the coast of the United States.
2 Vibrio vulnificus The symptoms of V. fever. parahaemolyticus the estimated annual incidence rate is very low. Healthy individuals are susceptible to gastroenteritis. cholerae non-01 is a common bacterial cause of molluscan shellfishassociated illness. vulnificus are fever. They are sensitive to cold. are particularly susceptible to primary septicaemia. 56). The organism attaches to the intestine and release of a toxin is suspected.MOLLUSCAN SHELLFISH Control of V. In one study. 3.0.000 population/year (8. cholerae non-01 (51). Control of oyster-associated 67 . Again. the organism has been found in the Gulf of Mexico. It causes diarrhoea. in the range of 0. cholerae non-01 has been associated with septicaemia. gastroenteritis. 3. cholerae non-01 is ubiquitous in the estuarine environment. 46. 46. 49). parahaemolyticus. V.7 days. diabetes. V.5. 55. those who have liver disease.3 Vibrio cholerae non-01 Illness caused by V. which have been implicated in shellfish-related disease outbreaks (31). with a fatality rate of 50% for individuals with septicaemia. cholerae. lasting 6 . vulnificus. Onset of symptoms occurs in 48 hours and can be severe. leukaemia. The widespread distribution of this bacterium is comparable to other studies in the Gulf and Atlantic Coasts (52. however. and vomiting. The bacterium is widespread in estuarine waters particularly in warm waters of >20 °C. 23 of 24 major USA-West Coast estuaries were positive for V. 58). or primary septicaemia. 48). Hazards from this bacterium can be controlled by thorough cooking of shellfish. The bacterium is an estuarine species that causes wound infections. vulnificus is by rapid refrigeration during warm weather months. Primarily. and nausea within 24 . Illnesses are usually associated with the consumption of raw oysters as described in Table 3. and preventing cross contamination once the product is cooked. vulnificus causing septicaemia and death has been associated with the consumption of oysters (8. and rapidly become inactive at low storage temperatures. The occurrence of V. 53. primarily in immunocompromised individuals with a mortality rate exceeding 50% (50). It is one of the most severe foodborne infectious diseases. 57. cholerae non-01 is generally a less severe gastroenteritis than that caused by V. Oysters are the most hazardous food associated with V. All isolates are considered pathogenic. cirrhosis.4 . or immunosuppression.II (8. High-risk individuals. nausea.5. parahaemolyticus growth in shellfish meats is temperature dependent. Death has been reported to occur within 36 hours from onset. Control of V.8 case/100. abdominal cramps.48 hours of onset.1. like V. these individuals are advised not to eat raw shellfish. Inadequate refrigeration will accelerate the growth of V. 54. V. chills.1. the organism has also been found in cooler temperatures from estuaries of Washington and Oregon (47).
5%) and 2 . including V.5. and nausea. 3. cholerae non-01 illnesses is by cooking seafood thoroughly. Serotype Ogawa biotype El Tor and 0139 are responsible for epidemics in South American. nausea. cholerae 01 in molluscan shellfish. mimicus.2 Campylobacter jejuni C. is the same as previously discussed for the non-01 strains. abdominal pain. It causes diarrhoea. fluvialis. respectively. V. They are common marine bacteria. 60).1. processors should know the product source. whereas excellent sanitation facilities in developed countries are responsible for the near eradication of epidemic cholera. fever.FISH AND SEAFOOD V. In addition.1. and illness lasts 7 . 3. nausea. and southern Asia including India and Bangladesh. and V. or cross-contaminated shellfish (57.5.10 days. (8. these organisms cause gastroenteritis lasting 24 .10% carbon dioxide for optimal growth conditions. vomiting. and harvesting of oysters in colder months when Vibrio counts in water are the lowest (59). There are several other Vibrio spp. imported product from a country experiencing an epidemic. parahaemolyticus and V. The presence of these strains appears to be in low numbers and non-virulent (59). undercooked. Poor sanitation and contaminated water supplies will spread the disease.48 hours with abdominal pain.5 Other Vibrios spp. cholerae 01 is transmitted by faecal contamination of food or water. acidity and heat. diarrhoea. Death may occur after severe fluid and electrolyte loss. The frequency of infection of these organisms is no different from other Vibrios. 68 . drying. salinity. V. In general.g. headache and fever. jejuni requires reduced levels of oxygen (3 . cholerae 01. 62.S. Illness can include abdominal cramps. and shock. particularly in oysters. In developing countries. low pH. 3. vulnificus. Illnesses are associated with the warmer months. It is considered a fragile and sensitive organism to environmental stresses such as oxygen. V. The control of V.5. dehydration. There is evidence of strains of V. however. Onset of disease is 6 hours to 5 days. 63). Control of infection from these organisms is no different from the other Vibrios. Gulf coast environment.4 Vibrio cholerae 01 and 0139 Symptoms of epidemic cholera vary from mild watery diarrhoea to acute diarrhoea with characteristic rice water stools. associated with shellfish-borne illness outbreaks. Onset of illness is from 3 to 5 days after ingestion of contaminated food. hollisae. and the transmission of the disease by consumption of raw. e. the pathogenic severity is lower than V. widespread in both polluted and unpolluted waters. 61. preventing crosscontamination of cooked seafood. cholerae 01 establishing in the U.
and have been implicated as the causative agent in illnesses from the consumption of raw oysters and clams (68. 72. they survive well at refrigerated temperatures. 84.3 Other organisms of concern 3. hollisae Infective dose varies with host susceptibility and virulence of the strain. C.1 Aeromonas hydrophila group and Plesiomonas shigelloides A.500 cells/gram of Campylobacter may cause illness (64. Although.MOLLUSCAN SHELLFISH TABLE 3. 70. however. although their role as a pathogen is highly controversial and circumstantial. ranging from 0. C. The seasonality of 69 . mimicus. Proper care should be taken if the finished product is to be packaged in modified-atmosphere packaging (MAP). Campylobacter spp. V. vulnificus. have been isolated from shellfish beds throughout the world (67. This finding is significant in that C. hydrophila group (75. V. cholerae 01 and non-01. jejuni begins in the harvest of shellstock from approved waters. Recently. the incidence of Campylobacter infection is low from the consumption of molluscan shellfish. 65. from Pacific shellstock oysters purchased from a retail grocery store (74). control of C. shigelloides (79. 3. however. They have been implicated in limited shellfishassociated gastroenteritis cases (8. Sixteen of 28 participants became ill with campylobacteriosis after ingesting raw clams. 66). To reduce the risk of shellfish-borne campylobacteriosis. 82. jejuni does not multiply in shellstock at refrigerated temperatures. 69.4 to 114 cells. whereas at 10 ºC Campylobacter survive only for 5 days (71). In one case. 85). fluviallis. 76. parahaemolyticus.5. 81. 68.3. 83. 73). V. of Cases 4 362 0 1 0 a Includes V.a (8) Species Hard clams Oysters Soft Clams Mussels Scallops No. 77.II Molluscan shellfish associated outbreaks caused by Vibrio spp. 78) and P. of Incidents 4 279 0 1 0 No. jejuni survived for over 20 days in market-shucked oysters stored at 4 ºC. jejuni were recovered. infection occurred among participants of a firemen's banquet. and V. and their immediate proper refrigeration.5. 71). 80) are ubiquitous in the marine environment. Campylobacter appear to survive well in oysters during storage. Campylobacters do not survive well in the environment due to the organism's lack of resistance to environmental factors. Human feeding studies suggest that about 400 . V. the organism is being continually inoculated into the environment by waterfowl and farm run-offs and thus eventually contaminating shellfish growing waters.
parahaemolyticus Staph. Pathogen monitoring of Selected High Risk Foods Domestic including: milk. Test Standard Plate Counts E. FDA/ Administrative Guidelines.6 Published Microbiological Criteria The published microbiological criteria below are included as a guide. Note: Although molluscan shellfish are not included in the selected high risk food. Control of these organisms in shellfish products is limited. hydrophila group stored at refrigeration temperatures. 70 . after 7 days storage of shucked oysters at a shellfish processors’ cooler kept a 5 ºC Aeromonas counts ranged from 500.000 MPN per gram1 negative for staphylococcal enterotoxin or when the viable MPN count is <10.000 cfu/g1 <230/100 g2 <1000 per gram. FDA established action levels or levels of concern for certain microbial pathogens in molluscan shellfish. jejuni V. They survive well in shellstock or shucked oyster meats held at refrigeration temperatures. 87) of the outgrowth of A. non-dairy frozen desserts. 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. FDA/Administrative Guidelines 3 Source (91). prepared sandwiches. dried milk/whey drink powders. Enforcement action is considered on a case-by-case basis taking into account all the factors associated with the specific situation. and where indicated are intended to be used for official control purposes. coli Enterotoxigenic E. soy products.0001 negative for the presence3 Under review4 Source (89). action is considered on a case-by-case basis taking into account all the factors associated with the specific situation.000/gram (29).000 to 3. The Agency will consider enforcement action against the shipment of molluscan shellfish if the levels exceed as stated. 2 Source (90). They are easily destroyed with mild cooking temperatures. being isolated more often during the warmer month. suggesting that these individuals are at higher risk when consuming raw or partially cooked shellfish containing these bacteria. cholerae V. In one study. Similar results were seen by other investigators (86. vulnificus in molluscan shellfish. coli Salmonella V.FISH AND SEAFOOD Aeromonas and Plesiomonas is similar to the Vibrios. aureus C. vulnificus 1 Acceptable Levels in Molluscan Shellfish <500.600. 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). 3.
1964. 1995. Wiley-Blackwell. Ward D. 1963. in Microbiology of Marine Food Products.H..R. 71-6. 33643. 6. Hackney C. 1982.A. 105-8.. American Journal of Hygiene.D. References The Phylum Mollusca. 7. 1968. water and sediment from Galveston Bay. 1926. Liston J. 755-76.S.K. Tubiash H.M. 1984.F. Applied Microbiology. 2. Tall B. Veliger. 1940.K. Old H. 29. Organisation for Economic Co-operation and Development. 15.R. 58.. Lovelace T. 1975.. Microbiology of shellfish: bacteriological study of the natural flora of pacific oysters (Crassostrea gigas). Scanning electron microscopy of Cristispira species in Chesapeake Bay oysters. American Journal of Public Health. Walker W. Saunders Company. Viability and release Salmonella charity and Escherichia coli from oyster faeces. Galtsoff P.MOLLUSCAN SHELLFISH 3. Eliot C..E. 26. Smith F. Eds.W. 46. Jr. American Journal of Public Health. Bacterial flora of the market oyster. 544-8. The American Oyster.C. Washington D.. Rippey S. 11. 19-34. 3.. Applied and Environmental Microbiology. 9.R.7 1. Galtsoff P. Schmidt C. Infectious diseases associated with molluscan shellfish consumption. Government Printing Office. 6. Vanderzant C. Effects of harvesting waters and storage conditions on yeast population in shellfish. 8. Cook D. Gill S. 1991.R. 1973. Microbial flora and level of Vibrio parahaemolyticus of oysters (Crassostrea virginica). 8.C.. Bernard F. 33-6. 10. 7.S. 17. Villee C. 5. 633-40. Clinical Microbiology Review. Lee J. U. Biology of the oyster in relation to sanitation. 14.. USA.S.A. Eds.R. 71 . Occurrence of the spirochaete genus Cristispira in western Canadian marine bivalves. Journal of Food Protection. 42. Nauman R.L.. Colwell R. 16. Journal of Milk and Food Technology. in General Zoology.E. Rowse A. Ray S. A typhoid fever epidemic caused by carrier bootlegging oysters.W. Philadelphia. Microbiological characteristics of Dungeness crab (Cancer magister). Colwell R. 1960. 72-8. 13. 447-52. 1970. 1983.S.. Proceedings of the National Shellfisheries Association.B. 30. Pfeifer D. 4. Applied Microbiology. 44. Quantitative and qualitative commensal bacterial flora of Crassostrea virginica in Chesapeake Bay. Microbiology of bivalve molluscan shellfish. 245-7.A. New York. 36. 419-25. 231-51. 1936. W.R. Fate of enteric bacteria in estuarine sediments and oyster faeces. Ed. 13. Cook D. Fleet G. Thompson C.N. Applied and Environmental Microbiology... Jr. 64. 30. in Multilingual Dictionary of Fish and Fish Products.J. 1981. Fish and Wildlife Service. 12. Hood M. 1994. Van Nostrand Reinhold. in Fishery Bulletin 1964 No. Journal of the Mississippi Academy of Sciences. 104-9. 82-7.
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4. 6-9 September 1983.E.. 64. (jejuni.A. Abeyta C. Levine M. Wilson T.L. Infective dose of Campylobacter jejuni in milk. Rowe B. Pearson A. 1983..MOLLUSCAN SHELLFISH 62.. Kaysner C. 257.K.E.A. 1584. 1995. Eds. Alam A.R. Phenotypic and genotypic characterisation of urease-positive thermophilic Campylobacter (uptc) isolated in N. Wareing D.. Incidence and survival of thermophilic campylobacters from shellfish growing water: media evaluation. in Campylobacters.. Morris G. Huges T. 75 .M. 99. Public Health Laboratory Service. 11... 1981. Matsuda M. 56.. Deeter F. Clements M.W.. 68. Feldman R.. Journal of Infectious Disease.. 282. Campylobacter jejuni in a Washington State shellfish growing bed associated with illness. Davis B..A..G. Johnston J. Wilson T. Hughes T. Davis J. 13.. from patients with diarrhoea.S. Wilson I. Clements M.. 101-4.D. 51-64.. Stott R. 1987..A..E.. 1996.. 66. Arumugaswamy R. S41. Blake P... Ireland.. Huq M. Brenner D.. a newly recognised Vibrio species. 472-9. Journal of Clinical Microbiology.P. Part 1 Abstracts: Shellfish as a possible source of thermophilic Campylobacter spp. in Campylobacter II: Proceedings of the Second International Workshop on Campylobacter Infections..G. Griffin M. Plenum Press.H. Skirrows M. Ketley J.. Dalley E.. 67.D. 70. Abeyta C. Kaysner. Annals of Internal Medicine. 621-4.G.X. 69. Jr. New York. Campylobacter gastroenteritis associated with raw clams. Moore J..L. Brussels.M.. 71.G.R. infections in man. 72. 63.J. 1980. Wekell M... Wekell M.E..J. London.G. 1980. 1991. Ketley J. 1983.R..P.M. Jr. 169-71.G... Murphy P. Newell D. Journal of Food Protection. Levine M. Newell D. Helicobacters and Related Organisms. Microbial Ecology in Health and Disease.. Studies of Campylobacter jejuni infection in volunteers. 80. New York. 607-9. Hunt J.F.. Blaser M. The occurrence of Campylobacter jejuni and Campylobacter coli in Sidney rock oyster. Blaser M. A method for isolation of Campylobacter spp. in Campylobacters.K.S.M. Murphy P. 65. Journal of Rapid Methods and Automation in Microbiology... 73. Eds..M. Jones D.M.G.A. Robinson D.A. Jr. 4. coli and lari) from shellfish and the marine environment. British Medical Journal. Disease from infection with Vibrio mimicus. 157. Wilson I.K.A.A. Eds. 256. C.M.. Moore J. 1993.J. Isolation of vibrio-like group.R.M. Helicobacters and Related Organisms. Wareing D.I. Experimental Campylobacter jejuni infection in humans. Kaysner C. Abeyta C. Journal of the Medical Society of New Jersey. 1988. EF-6. International Journal of Food Microbiology.J. 323-5. Austin S. 4. Feldman R. Shandera W. Proudford R.. Plenum Press. Black R. Fitzpatrick M.R. 1996.B.M.A. Black R.
M. Levine M.. 1990. 79. 998-1001.A. 1981.L. 87. Tall B..A.B.. 81. 75. vibrios and Aeromonas hydrophila. 731-8. 1986.. Journal of Food Protection.A.FISH AND SEAFOOD 74. Baker R.A..M. Cook D.W. Sullivan J.. Hood M.. Journal of Rapid Methods and Automation in Microbiology. Kaysner C. 105. 598-601. Herrington D. 1988.. 47.. Vandepitte J. Prevalence and distribution of Aeromonas hydrophila in the United States. 78. Biotyping and virulence factors in clinical and environmental isolates of Aeromonas species. GA. 47. Hickman-Brenner F. 475-9. Koburger J.D.. 1989. Stelma G. 1978.M.. Oyster borne disease outbreaks in St. Trost P. Indicator bacteria and Vibrionaceae multiplication in postharvest shellstock oysters...C.C. Hunt J. Peterson D. Burke V. Wekell M. Singelton F. 80. Journal of Applied Bacteriology... Ruple A.A. 27..M. 1985. 1997. Tenge B. 449-57. Beaman J. Miller M. Applied and Environmental Microbiology. Herrington T.J. Van Damme L.H.L. 85.. 7-12. Holmerg S. 1989. 1980.R. Petersburg.J. Applied and Environmental Microbiology.P. 979-85.. Effects of processing and storing oyster meats on concentrations of indicator bacteria. Partridge K.M. Kaysner C. Annals of Internal Medicine. epidemiology. 48. Jr... Lockman H. III. Aeromonas hydrophila: ecology and toxigenicity of isolates from an estuary. 76. Infection and Immunity.. Cooper M. 849-55. Fliermans C. 36. Jr.A. Journal of Food Protection.F. Journal of Food Protection.. 223-47.R..W. 343-9. 39. Colwell R.D. 359-77.. 5. Report (1/13/84). Kaper J.. Farmer J.A. 1984.N.. Bark D. Abeyta C. 1146-9. Kelly M. Tzipori S. Plesiomonas shigelloides: An opportunistic food and waterborne pathogen.D.. Wekell M.. Blake P. Seidler R. 77. Hirsch R. Jr. Recovery of Aeromonas hydrophila from oysters implicated in an outbreak of foodborne illness...L. 52... Plesiomonas enteric infections in the United States. 1984. 55. Esch G. 83... Hazan T. 53. Atlanta. Fla. Kaysner C. Kain K. 82.. 84..M.T. and treatment of Plesiomonas shigelloides diarrhoea. Wekell M. Frequent isolation of Edwardsiella tarda and Plesiomonas shigelloides from healthy Zairese freshwater fish: A possible source of sporadic diarrhoea in the tropics. Department of Health and Human Services/Public Health Service/Food and Drug Administration. 86.A.. Journal of Food Protection. Journal of Clinical Microbiology.. Journal of Food Protection. 52. 1987. Robinson J. Abeyta C. Stott R.. 50. Gracey M. The use of bacterial membrane fractions for the detection of Campylobacter species in shellfish. Incidence of motile aeromonads from United States west coast shellfish growing estuaries. 76 ..J. Abeyta C. Clinical features. 690-4. Robins-Browne R. In vitro and in vivo pathogenicity of Plesiomonas shigelloides.J. Wachsmuth I.M.K. Applied and Environmental Microbiology.W.
Part I Sanitation of Shellfish Growing Areas. Sherlock C. Ed. Aspen Publishers Inc. Seafood Safety.W. Government Printing Office . 2000. Food and Drug Administration Compliance Program Guidance Manual.C. U.. U. Food and Drug Administration..1987. 90. Washington D. Food and Drug Administration. Lancaster. 63-8. 77 .H.S.S.. 7. U.. Does Aeromonas hydrophila preferentially colonise the bowels of patients with hematologic malignancies? Diagnostic Microbiology and Infectious Disease..M. Ed.S. U. in U.S. Food and Drug Administration.. Inc.. Food and Drug Administration.R. Collette R.E. U.W.030. 89. Gaithersburg. Smith J. 1997. 1995. Fish Inspection. Food and Drug Administration. Washington D. Ed. Quality Control.8 Further Reading Ahmed F. Food and Drug Administration Compliance Policy Guides. mussels. 1995. 1987.S. Lund B. in National Shellfish Sanitation Program Manual of Operations.12). Chapter 5: Foods. fresh or frozenadulteration by bacteriological contamination Sec. Clams. U. The Microbiological Safety and Quality of Food.. Washington D. 560. oysters. Pittsurgh. Food and Drug Administration. Gould G.S. Slavin J. 1991.600 (CPG 7119.MOLLUSCAN SHELLFISH 88.. and HACCP: A Global Focus.E. National Academy Press.S. Food and Drug Administration. Technomic Publishing Co. Martin R. Food and Drug Administration.C. 3. Baird-Parker T. Program 7303. 91.C.L.A.S. Burdge D.Pathogen monitoring of selected high risk foods (FY 88/89). in U.C.
5 and 1.5 and 1. Parapeneopsis spp.. e. and available in 0. lobsters. Breaded shrimp and scampi are battered and coated in breadcrumbs and sold frozen to the consumer for cooking prior to consumption. prawns and scampi are temperate species commonly eaten by man and include members of the families Nephrops norvegicus (scampi). with shells and heads on or off are usually packed. and frozen in 1 and 2. lobsters and crabs have ten legs.Ph. crayfish and shrimp (1). CRUSTACEAN SHELLFISH Linda Nicolaides. the front pair ends in claws.4. Shrimp can be purchased in the raw or cooked state in a variety of forms: Entire shrimp. Pandalus borealis (deep-water shrimp). with additional water to form a glaze. Such products are usually only partially cooked before freezing.5 kilo pack (boxes or bags). M. Peeled ready-to-cook shrimp are raw. and Trachypenaeus spp. jointed limbs and a chitinous exoskeleton.. 79 . Pandalus montagui and Crangon crangon (inshore shrimp). Peeled and cooked shrimp are ready-to-eat after controlled thawing.0 kg polythene bags.1 Definitions Crustaceans are invertebrate animals and represent a class of the phylum Arthropoda. Shrimps. They possess a segmented body. Species eaten from tropical waters include Penaeus spp. peeled product. crabs. packed and frozen in 0..g.0 kg polythene bags. (deep-water shrimp). Crayfish. and are usually aquatic. FRSPH Ethical Trade and Food Management Group Natural Resources Institute University of Greenwich Central Avenue Chatham Maritime Kent ME4 4TB United Kingdom 4.
g. their feeding and living habits. Poor hygienic procedures will increase the probability of contamination with bacteria of public health significance. with the increasing demand for ready-to-eat/cook crustacean products.2 Initial Microflora As with finfish. Microbiological risks associated with crustacean shellfish increase in proportion to the degree of handling that the product undergoes. which means that the haemolymph can be a reservoir of bacteria. e. the potential exists for crosscontamination from both the processor and from the environment. lobster and crayfish are available for the consumer owing to the growing demand for ready-to-eat products. this presents more favourable conditions for growth by both spoilage and pathogenic bacteria. For example. or frozen crab. However. boiled to kill the crustaceans and cook their meat prior to consumption. traditionally. The structure of the circulatory system of crabs is not closed. hence. shrimp die immediately after capture. the season. for crabs and lobsters. the level and type of the initial microflora of crustacean shellfish will reflect a combination of factors. lobsters and crayfish are traditionally kept alive after capture and then cooked to kill immediately before consumption. Shewanella putrefaciens and 80 . If these products are subjected to temperature abuse. In comparison. Spoilage begins following death once the immuno-response system of the crustacean shellfish fails to function. stored and transported as frozen product.FISH AND SEAFOOD Crabs. there is a growing market for fresh lobster tails or cooked crabmeat in crab shells. Cooked and chilled. Control measures to prevent these potential hazards occurring include training and supervision of operatives as well as Good Hygienic Practices (GHP). packed into polythene bags. the flora will change depending upon the methods of handling and/or the environmental conditions to which they are exposed.g. and the temperature and quality of the waters in which they exist. it is important that shrimp are handled and iced correctly in order to keep them in as fresh and uncontaminated a state as possible. Following capture or harvest. so that spoilage may occur during landing and transportation to the processing plant. spoilage bacteria are of little importance as. Spoilage bacteria associated with temperate crustaceans include Pseudomonas spp. tropical waters. Pasteurised crabmeat is edible meat removed from the exoskeleton by hand. As with finfish. e. particularly for members of the genus Vibrio (2). storage above the recommended storage temperature. during the peeling of cooked shrimp. only live carcasses are processed. lobster tails and cooked crabmeat presented decoratively in their shells. 4. particularly after it has been cooked. The microbial flora of crustaceans from temperate waters differs from that of crustaceans from warmer. the geography of the area they are captured from or are grown in. which include the environment from which they have been captured or harvested..
Shrimps can be processed whole. with “head on” or “head off” whereby the edible tail is removed from the head. Vibrio spp. These animals can survive out of water if the surrounding environment is kept cool and damp. corynebacteria. Alcaligenes. glazed and frozen (see Figure 4.for example. peeled and cooked. especially if they are stored in ice or iced water. the genera associated with tropical species include Enterobacteriaceae.3. up to 16 days for tropical species compared with 8 . or are found in brackish water around the shores. gills and thorax. so they are either sold on the local market or are processed in local processing plants. 4. After capture and transport to the processing plant. thus.. chilled 81 . uncooked prawns and shrimp is presented in Figure 4. they are washed.1). Shrimp will then be glazed and frozen in a raw state.10 days for temperate water species (3). Scrupulous hygiene is important at this stage to prevent the cross-contamination of the shrimp tails. the rate of deterioration of the crustacean shellfish by these organisms is reduced. They are then taken to market alive in either holding tanks. or transported directly to processing plants. chilled sea water. The latter process can expose the shrimp muscle to incidents of cross-contamination if GHP are not followed. In contrast. by removing the head and thorax a source of enzymes is eliminated that will prevent the occurrence of black spots. and will be dependent upon whether the crustaceans were harvested from cool or warmer waters. In comparison shrimps die quickly after capture. lobsters and crayfish are harvested from cages or pots containing bait. however. As with finfish. Pseudomonas spp.3 Processing and its Effects on the Microflora Crabs.CRUSTACEAN SHELLFISH members of the Acinetobacter-Moraxella group. When the shrimp arrive at the processing plant they are washed and then sorted into size. from the time of capture and processing. The claws are usually bound together to prevent the animals fighting and causing damage to each other. 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. Shrimp might be further processed with the shell on.. As the muscles of crustacean shellfish decompose rapidly after death dead animals are rejected. A large proportion of shrimps are produced by aquaculture. there is evidence that the shelf life of tropical crustaceans is longer than that of temperate-water species stored correctly in ice . bacterial contamination and spoilage are inevitable. or with the shell and central vein removed. The rate of spoilage will be a function of time and temperature. Hence. and Achromobacter spp.1. graded according to size. or cooked.1 Prawns and shrimps An example of the process used for preparing frozen. whether GHP are in place. 4.. Micrococcus spp.
frozen uncooked prawns 82 . After cooking. 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. 4.FISH AND SEAFOOD frozen. 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. spices and cooking batter. and packed to meet the customer’s specification.1. particularly the pathogens Bacillus cereus and Clostridium perfringens originating in the flour. Breaded products introduce an added risk of spore forming bacteria. Flow diagram . Fig. Good Manufacturing Practice (GMP).
However. slightly iodine smell. the tightness of limbs will indicate whether they were killed by immersion in boiling water or. An offensive odour is also produced. wet and ‘soapy’ or ‘jammy’ to the touch. or in hermatically sealed cans that have undergone an appropriate heat treatment. contaminating microflora may include enteric bacteria. spoiling prawns and shrimps have shells that are soft. bright and dry appearance. 83 . putrefaciens and Pseudomonas spp. with visible decomposition or green discoloration seen through the membrane exposed by the dropping tail. immediately prior to boiling.2 Crabs and lobsters Crabs and lobsters should be alive when boiled or sold. 4. since crabs and lobsters are traditionally consumed immediately after cooking. the joint between the tail and carapace is open and the limbs hang loosely. If boiled.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. dull and sticky. In comparison. and have a sweetish. there is no associated spoilage problem. whilst boiled crustaceans have a pleasant slightly sweet smell. the meat is either repacked and sold in the shells.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. If hygienic procedures are not followed. there is a new hazard of contamination of the cooked meat by workers during its physical removal from the exoskeleton. 4. 4. viruses or parasites.4. bacteria of public health significance. there is no indication of life. If the tail drops. Heat is produced if the shrimps decompose in a confined space.2 Cooked crabs and lobsters As indicated earlier. The shell will be soft. for crabs. 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.3. or packaged and processed. they include Sh.CRUSTACEAN SHELLFISH 4. with the growing demand for ready-to-eat products from crab and lobster meat. putrefaciens.4. With spoiled crabs and lobsters. The shells have a crisp. indicative of protein degradation and the growth of the principal spoilage bacterium Sh. and a strong ammoniacal odour is produced.
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. The presence of L. Salmonella cannot be completely removed from raw shrimp during processing. However. nearly 30 years elapsed before the potential hazard of L.5. monocytogenes in contaminated crustacea was recognised in the United States of America (USA) when the pathogen was isolated from Mexican frozen cooked crabmeat (10). 6. One of the earliest accounts of the presence of Listeria monocytogenes in crustaceans was by Russian workers in 1959 (9). An import alert was put in place in June 1987. The pathogen was later identified in frozen raw shrimp (13) and lobster tails (14) . even if GMP is followed. monocytogenes in cooked ready-to-eat crustaceans is recognised as a potential risk to selected groups of individuals: the very young and 84 . in compliance with the Food and Drug Admistration’s (FDA) ‘zero tolerance policy’ for L. a common pond fertiliser used in aquaculture. Results of a survey carried out in England and Wales between 1987 and 1989 demonstrated that L.cooked prior to consumption. the first in a series of Class I recalls was issued in May 1987 to retrieve nearly 4 tonnes of tainted crabmeat (11). cooked and smoked fish. Although foodborne illness had not been associated with the product. 16).FISH AND SEAFOOD 4. which required automatic detention of all Mexican frozen crabmeat that tested positive for Listeria and Escherichia coli (12). Further surveys identified Listeria spp. The above results prompted the FDA to develop a compliance programme for the pathogen. and to increase testing of many other domestically produced seafoods under the General Pathogen Surveillance Programme (14. shrimp and shellfish (11). Following this. shrimps and cockles sold (17). monocytogenes was absent from 40 samples of cooked prawns. in 24. Aquatic birds and the hands of workers have also been identified as sources of the pathogen. 5.5% of frozen raw shrimps from different countries (13). later surveys in 1990 recovered the pathogen from retail raw.5 4. Moreover. monocytogenes in cooked ready-to-eat foods. fish fingers.5. The genus Salmonella contains a wide variety of ‘species’ pathogenic for man or animals. 4. and usually for both. as well as for Salmonella.1 Pathogens: Growth and Survival Salmonella spp. often contains Salmonella and other enteric pathogens (8) and has been demonstrated to contaminate crustaceans growing in these environments. animal manure. in domestic/imported shrimps (15). They are mesophilic microorganisms usually residing in the intestinal tracts of human beings and warm-blooded animals.2 Listeria spp. Hence. although efficient cooking will quickly eliminate it (6). 7).
4. Crustaceans have been linked with food intoxications attributed to Staph. Researchers have demonstrated that inoculated shrimp supported toxin production by C. aureus because of the ubiquity of this organism in the processing environment. 25). V. botulinum type E at 10 °C but not at 4 °C (20). 4. Control should be achieved by promoting GHP and staff training programmes. no incidence of foodborne illness caused by L. The pathogen has resistant spores. particularly crabmeat (18. which can become colonised by V.5. although more recently the non-O1 type has also been demonstrated to cause foodborne illness in crustaceans. The former.5 Vibrio spp. Vibrio parahaemolyticus and Vibrio vulnificus are species that have been reported to be associated with raw crustaceans harvested from estuarine waters (21. combined with the degree of handling of these types of product. e.5. 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. so is able to survive heat treatments that eliminate vegetative cells. as part of the natural bacterial flora. V.g. about 50 . 4. whilst boils and cuts are also common sources of the pathogen. although contamination by types A and B may occur from the environment.3 Staphylococcus aureus Staph. cholerae. was associated with seafood consumption. aureus on their hands. the causative agent of cholera. which was eventually attributed to a ship discharging its bilge into the coastal waters of Peru. the pathogen can grow at temperatures as low as 3 °C. therefore. The main source of contamination is the hands of processing personnel. 24. 22) as well as from undercooked crustacea when harvested from contaminated waters (23. is widely distributed in aquatic environments. pregnant women and the immuno-compromised. Once the competing microflora has been removed by a heat treatment. Vibrio cholerae. botulinum type E is commonly associated with fish and fishery products. cholerae non-O1. However.5.CRUSTACEAN SHELLFISH old. An epidemic that began in South America in 1991. cholerae O1 and V. Crabs are covered in a chitin exoskeleton. To date.4 Clostridium botulinum C.70% of healthy individuals carry toxin-producing strains of Staph. especially in products that have been vacuum packaged. 19). cholerae is divided into two groups. monocytogenes has been attributed to crustaceans. 85 . pasteurisation and cooking. GMP should be followed in the processing plant to ensure that the final product is held at 3 °C. aureus is not a member of the intrinsic flora associated with crustacean shellfish.
30 MPN/g) (27). whilst 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. particularly trematodes (6).7 Aeromonas hydrophila and Aeromonas sobria A.FISH AND SEAFOOD and caused major economic losses to South American countries that export shrimp to the United States. Parasitic infections are controlled by either cooking infected material prior to consumption. been implicated in a case of foodborne infection associated with crustaceans. vulnificus is an invasive and lethal pathogen associated with wound infections. to date. occurs in crustaceans from warm environments. they have not been confirmed to be causal agents of foodborne illness associated with crustaceans (28). However. hydrophila is able to grow well at refrigeration temperatures (0 . hydrophila and A.5. 4. Quantitative levels were below limits of detection in all cases (<0. Norovirus. coli.g. but has the potential to cause fatal foodborne illness (26). or freezing at a temperature of -23 °C for 7 days (30.8 Parasites The use of animal excreta in aquaculture has also been associated with foodborne parasitic infections of crustaceans. 4. Hepatitis A virus presents the most serious hazard. V.9 Viruses Crustaceans grown in waters contaminated with sewage may become carriers of viruses such as Hepatitis A. a marine halophile.8% of samples contained C. However. Of these. 4. Astroviruses and non-A. hydrophila has been isolated from prawns (29).5. in a survey carried out in the United States between 1977 and 86 . sobria are known to be fish pathogens and are commonly isolated from ponds used for aquaculture. South America. however. V. parahaemolyticus. 32). non-B Hepatitis viruses (31). It is more commonly implicated in countries where crustacean shellfish are eaten raw. e. Caliciviruses.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.5. jejuni has not.5. although A.6 Campylobacter jejuni and Campylobacter coli C. jejuni was isolated from 36 (15%) of 240 samples of freshly hand-picked blue crab (Callinectes sapidus) meat. 4. A. 31. Japan. The organism is widespread in estuarine waters and is a common isolate from harbour water in warm climates. the potential hazard exists as C.
Therefore any microbiological criteria need to be part of this HACCP-based system to verify that the system is under control.CRUSTACEAN SHELLFISH 1981. 38. A summary of microbiological criteria for crustacean shellfish is listed below.47% of outbreaks attributed to Hepatitis A virus were attributed to a foodborne origin (33). which results in a persistent and apparently permanent loss of short-term memory (36). or chlorine concentrations similar to those found in drinking water.5. 39). The toxin causes amnesic shellfish poisoning (ASP) when consumed. from farm to fork (37. 4. No foodborne incidents have been attributed to this source to date. which could be transferred to the meat during unskilled removal from the shell (35). They can survive low pH levels. although it is one of the European Union (EU) legislated allergens. only 0.10 Allergens Allergens have not been reported to be a significant hazard with crustacean shellfish. 87 . 4.6.6 Published Microbiological Criteria The global legislative requirement for controlling the safety (and quality) of crustacean shellfish is to apply a preventive.5. 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. produced or imported into the European Union (EU). Provisions laid down concerning the use of additives would also apply to these types of products. The risk of such hazards is far higher with fin fish and molluscan shellfish. 4.1 EU Legislation Microbiological standards for cooked crustaceans and 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. Norwalk viruses can also withstand heating to 60 °C. are included in the Commission Regulation (EC) No. 4. 2073/2005. consistent with that in the stomach. Both Hepatitis A virus and Norovirus are stable pathogens and are relatively resistant to inactivation (34).
FISH AND SEAFOOD
22.214.171.124 Microbiological criteria applicable to the production of cooked crustaceans (Commission Regulation (EC) No 2073/2005 on microbiological criteria for foodstuffs) 126.96.36.199.1 Chapter 1 - Food safety criteria
L. monocytogenes Prepared crustacean products fall into the category described in the regulation as “Ready-to-eat foods available to support the growth of L. monocytogenes other than those intended for infants and for special medical purposes”. During capture and preparation there is a zero tolerance for the presence of L. monocytogenes on the product. However, to take into account the issues of natural habitats being contaminated with low levels of this pathogen there is a slight loosening of the regulation at the point of sale and for the duration of the products shelf life (refer to Table 9.I, Food Category 1.2). If the level of L. monocytogenes should be above this level then the consignment is considered to be unsatisfactory. Salmonella All cooked crustaceans and molluscan shellfish should be free from Salmonella at the point of sale and for the duration of their shelf life (refer to Table 9.I, Food Category 1.16). Should any sample test positive for Salmonella then the product is declared to be unsatisfactory. 188.8.131.52.2 Chapter 2 - Process hygiene criteria
Process hygiene criteria for E. coli and coagulase-positive staphylococci in shellfish and shucked products of cooked crustaceans and molluscan shellfish are laid down (refer to Table 9.II, Food Category 2.4.1). 4.6.2 International requirements
International requirements are based upon having a risk-based, preventive management system in place at all stages of the supply chain. The requirements for this preventive approach is described as follows (37): • Fish and fishery products should be prepared in plants certified by the local competent authority. All certified plants should comply with the GHP (39); • The fisheries industry should take responsibility, implement and maintain safety management systems based upon HACCP; • The national competent authority is responsible for the certification of fish processing and manufacturing plants, verification of effective systems, and
issuing of certificates of compliance for export products. This includes auditing and inspection programmes; • National surveillance and monitoring programmes should be in place to demonstrate that all identified hazards are under control, e.g. biotoxins, and to identify potentially emerging hazards; Within this requirement is the flexibility for countries and trading blocks to set specific standards required to deliver an equivalent requirement with those at national level. Such standards need to be science-based, especially if they are more stringent that those recommended by the Codex Alimentarius Commission. Verification that GHP have been followed during the handling and preparation of crustacean shellfish for consumption is usually done by assessing the level of E. coli and/or coagulase-positive staphylococci in the products. Legal requirements generally require that these groups of indicator organisms are present in levels <100 cells/gram of product. Pathogenic bacteria such as Salmonella spp. should be absence is 25 grams multiplied by the number of subsamples tested. 184.108.40.206 United States The FDA brought into force a Federal Mandate Seafood Rule in 1995 which forms the foundation for good practices required for the handling and processing of imported fish and fishery products into the U.S. The best practices include GHP and HACCP (37). All fish and fishery products produced in and imported into the U.S. after 17 December 1997 are required to demonstrate that HACCP principles were applied to their handling and processing. The USFDA has a zero tolerance for Salmonella in fish and fishery products; this includes farmed shrimp. The US FDA considers that the presence of pathogenic microorganisms in seafood is an indicator of adulteration (32). 4.7
Encarta 1997. Encarta 97 Encyclopedia World English Edition. Microsoft. The International Commission for Microbiological Specifications for Food. Fish and Fish Products, in Microorganisms in Foods 6. Microbial Ecology of Food Commodities. Ed. The International Commission for Microbiological Specifications for Food. New York, Kluwer Academic/Plenum Publishers. 2005, 174-249. Cann D.C. Bacteriology of shellfish with reference to international trade, in Handling, Processing and Marketing of Tropical Fish. Ed. Tropical Products Institute. London, Ministry of Overseas Development. 1976, 377-94. D’Aoust J.Y., Gelinas R., Maishment C. Presence of indicator organisms and the recovery of Salmonella in fish and shellfish. Journal of Food Protection, 1980, 43, 679-82.
FISH AND SEAFOOD 5. 6. 7. 8. 9. 10. 11. Fraiser M.B., Koburger J.A. Incidence of salmonellae in clams, oysters, crabs and mullet. Journal of Food Protection, 47, 343-5. Reilly P.J.A., Twiddy D.R., Fuchs R.S. Review of the occurrence of Salmonella in tropical shrimp. FAO Fisheries Circular No. 851, 1992, (FIIU/C851). Reilly P.J.A. Salmonella and Vibrio cholerae in brackishwater cultured tropical prawns. International Journal of Food Microbiology, 1992, 16, 293-301. Twiddy D.R. Antibiotic-resistant human pathogens in integrated fish farms. ASEAN Food Journal, 1995, 10, 22-9. Olsufjew N.G., Petrow V.G. Detection of Erysipelothrix and Listeria in stream water. Zhurnal Mikrobiologii Epidemiologii i Immunobiologii, 1959, 30, 89-94. Anon. First Listeria finding in crabmeat confirmed by FDA. Food Chemistry News, 1987, 29 (14), 38. Ryser E.T., Marth E.H. Incidence and behaviour of Listeria monocytogenes in fish and seafood, in Listeria, Listeriosis and Food Safety. Eds. Ryser E.T., Marth E.H. New York, CRC Press. 2007, 617-54. Anon. Mexican crabmeat, Greek pasta automatically detained. Food Chemistry News, 29 (19), 43. Anon. FDA checking imported domestic shrimp, crabmeat for Listeria. Food Chemistry News, 1987, 29 (24) 15-7. Archer D.L. Review of the latest FDA information on the presence of Listeria in foods. WHO Working Group on Foodborne Listeriosis, Geneva, Switzerland. 1988, 15-9. Anon. FDA to sample shrimp for Salmonella, Listeria. Food Chemistry News, 1988, 30 (36), 9-10. Anon. Monitoring of changes to listeriosis problem urged. Food Chemistry News, 1989, 31 (20), 13-7. Richmond M. Report of the Committee on the Microbiological Safety of Food. London, HMSO. 1990, 133-7. Swartzentruber A., Schwab A.H., Duran A.P., Wentz B.A., Read R.B. Jr. Microbiological quality of frozen shrimp and lobster tail in the retail market. Journal of Food Protection, 1980, 40, 765-9. Wentz B.A., Duran A.P., Swaartzentruber A., Schwab A.H., McClure F.D., Archer D., Read R.B. Jr. Microbiological quality of crabmeat during processing. Journal of Food Protection, 1985, 48, 44-9. Garren D.M., Harrison M.A., Huang Y-W. Clostridium botulinum type E outgrowth and toxin production in vacuum-skin packaged shrimp. Food Microbiology, 1994, 11, 467-72. Molitoris E., Joseph S.E., Krichevsky M.I., Sindhuhardja W., Colwell R.R. Characterisation and distribution of Vibrio alginolyticus and Vibrio parahaemolyticus isolated from Indonesia. Applied and Environmental Microbiology, 1985, 50, 1388-94.
12. 13. 14.
15. 16. 17. 18.
CRUSTACEAN SHELLFISH 22. Varma P.R.G., Iver T.S.G., Joseph M.A., Zacharia S. Studies on the incidence of Vibrio cholera in fishery products. Journal of Food Science and Technology, 1989, 26, 341-2. Madden J.M., McCardell B.A., Morris J.G. Jr. Vibrio cholerae, in Foodborne Bacterial Pathogens. Ed. Doyle M.P. New York, Marcel Dekker. 1989, 525-42. Twedt R.M. Vibrio parahaemolyticus, in Foodborne Bacterial Pathogens. Ed. Doyle M.P. New York, Marcel Dekker. 1989, 543-68. Oliver J.D. Vibrio vulnificus, in Foodborne Bacterial Pathogens. Ed. Doyle M.P. New York, Marcel Dekker. 1989, 569-600. Pearson A.M., Dutson T.R. Quality Attributes and Their Measurement in Meat, Poultry and Fish Products. Advances in Meat Research Series. Volume 9. London, Chapman & Hall. 1994. Reinhard R.G., McAdam T.J., Flick G.J., Croonenberghs R.E., Wittman R.F., Diallo A.A., Fernandes C. Analysis of Campylobacter jejuni, Campylobacter coli, Salmonella, Klebsiella pneumoniae and Escherichia coli 0157:H7 in fresh handpicked blue crab (Callinectes sapidus) meat. Journal of Food Protection, 1996, 59 (8), 803-7. Buchanan R.L., Palumbo S.A. Aeromonas hydrophila and Aeromonas sobria as potential food poisoning species: a review. Journal of Food Safety, 1985, 7, 15-29. Palumbo S.A., Maxino F. Williams A.C., Buchanan R.L., Thayer D.W. Starch ampicillin agar for the quantitive detection of Aeromonas hydrophila in foods. Applied and Environmental Microbiology, 1985, 50, 1027-30. Wekell M.M., Manger R., Colburn K., Adams A., Hill W. Microbiological quality of seafoods: viruses, bacteria and parasites, in Seafoods: Chemistry, Processing Technology and Quality. Eds. Shahidi F., Botta J.R. Glasgow, Blackie Academic and Professional. 1994, 196-219. Schantz J., McKerrow J.H. The dangers of eating raw fish. New England Journal of Medicine, 1989, 320, 1143-5. Ahmed F.E. Seafood Safety. Seafood Safety. Committee on Evaluation of the Safety of Fisheries Products, Food and Nutrition Board, Institute of Medicine. Washington D.C., National Academy Press. 1991. Blackwell J.H., Cliver D.O., Callis J.J., Heidelbaugh N.D., Larkin E.P., McKercher P.D., Tayer D.W. Foodborne viruses: their importance and need for research. Journal of Food Protection, 1985, 48, 717-23. Kapikian A.Z., Chanock R.M. Norwalk group of viruses, in Virology. Eds. Fields D.M., Knipe D.M. New York, Raven Press. 1990, 359-77. Horner R.A., Postel J.R. Toxic diatoms in western Eashington waters (US west coast). Hydrobiologia, 1993, 269/270, 197-205. Perl T.M., Bedard M.S.M., Kosatsky T., Hockin J.C., Todd E.C.D., Remis R.S. An outbreak of toxic encephalopathy caused by eating mussels contaminated with domoic acid. The New England Medical Journal, 1990, 322, 1775-80.
23. 24. 25. 26.
34. 35. 36.
FISH AND SEAFOOD 37. Ababouch L., Gandini G., Ryder J. Causes in Detentions and Rejections in International Fish Trade. FAO Fisheries Technical Paper 473. Rome, Food and Agriculture Organisation of the United Nations. 2005. Australian Government. Australian Quarantine and Inspection Service Hazard Analysis and Critical Control Point (HACCP). A Guide to Compliance with Export Control (Fish and Fish Products) Orders 2005. FAO/WHO. General Hygiene Requirements. Rome, Food and Agriculture Organisation of the United Nations. 2003.
Further Reading Spoilage and pathogens
Gram L., Huss H.H. Fresh and Processed Fish and Shellfish, in Microbiological Safety and Quality of Food Volume 1. Eds. Lund B.M., Baird-Parker A.C., Gould G.W. Maryland, Aspen Publishers Inc. 2000, 472-506. Ababouch L., Gandini G., Ryder J. Causes of Detentions and Rejections in International Fish Trade. FAO Fisheries Technical Paper 473. Rome, Food and Agriculture Organisation of the United Nations. 2005. International Commission on Microbiological Specifications for Foods. Fish and fish products, in Microorganisms in Foods 6. Microbial Ecology of Food Commodities. Ed. International Commission on Microbiological Specifications for Foods. London, Blackie Academic and Professional Press. 1998, 130-89. Sutcliffe P., Disney J. Handling, Processing and Marketing of Tropical Fish. London, Tropical Products Institute. 1977. Connell J.J. Control of Fish Quality. Oxford, Fishing News Books. 1995 Ward D.R., Hackney C. Microbiology of Marine Food Products. New York, Van Nostrand Reinhold AVI. 1991 Pearson A.M., Dutson T.R. Quality Attributes and their Measurement in Meat, Poultry and Fish products. Advances in Meat Research Series. Volume 9. London, Chapman & Hall. 1994.
4.8.2 Pathogens - general
International Commission on Microbiological Specifications for Foods. Microorganisms in Foods 5. Characteristics of Microbial Pathogens. Eds. International Commission on Microbiological Specifications for Foods. London, Blackie Academic and Professional Press. 1996. Varnam A.H, Evans M.G. Foodborne Pathogens - An Illustrated Text. London, Wolfe Publishing Ltd. 1991. Doyle M.P. Foodborne Bacterial Pathogens. New York, Marcel Dekker. 1989.
where the salt replaces moisture in the fish flesh. and the extracted moisture is allowed to drain away. to assure a salt concentration in the fish flesh that is appropriate for flavouring purposes prior to the use of other preservation techniques. drying and smoking processes. The immersion in concentrated brines for long periods is usually used for fatty species and for longer-term preservation. and marinating processes. Simon Derrick The Grimsby Institute Humber Seafood Institute Europarc Grimsby DN37 9TZ United Kingdom 5. their combinations and/or variations. drying. Wet salting or brining. Salting is both a method of preserving fish. both to preserve and to create a variety of new products. Usually. The processes take anywhere from a few hours to a few days or even months. This can be called ‘kench’ curing and is mainly used for non-fatty white fish. It includes the salting. rather than for preservation itself. fish is immersed for a few hours in a relatively low concentration brine. and a preliminary operation to some smoking. Fish curing is practised worldwide. and before significant spoilage takes place. CURED. and is sometimes known as ‘pickling’. alternating fish layers with a layer of salt. Fish can be salted in three different ways or their combination: Dry salting. SMOKED AND DRIED FISH Dr. split and opened out flat. 93 .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.5. The fish is headed. The fish is immersed in a brine solution in which different salt concentrations can be used according to the process. dependent on the country or the type of product. and salting is done by stacking the fish in layers.
Drying is the preservation of fish by removing the moisture content of the fish flesh by evaporation. Food products can be dried in three different ways: Air or contact drying. Longer shelf life is achieved in the products when salt is added. As the water is drawn out of the flesh by the salt a brine is formed. as concentrated brines are formed. This is a mixture of a dry and a wet salting process. thus delaying spoilage for a few days. Thus. Salt. It is not commonly used for fish products. Fish in contact with refrigerated plates freeze. Heat is transferred to fish from heated air or a heated surface. The product is dried by contact with heated surfaces or by radiation. or boiling it with dry salt and water. such as ‘gravad’ fish. depending on the type of product. under normal pressure. as well as other flavourings (sugar. The fish is mixed with dry salt but in a watertight container. prior to or during the boiling process. The principle of preservation of the first method is the inactivation of enzymes and killing the non-spore-forming organisms. Marinating or pickling. this was done naturally by exposure of the fish to the air. It has found limited application with fish. the preservative effect is due to the low pH used in the process. Salt-boiling. This is the common method of drying fish. Marinades are acidified brines. is used as a preservative instead of salt. evaporated water is removed by vacuum pump. thus making the product unsuitable for microbial growth.FISH AND SEAFOOD Dry / wet salting. When the process is running in a closed system. spices). are added to the marinade. The products are usually consumed raw. 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 . Freeze drying. thus reducing the water activity of the product. Included in this group are some lightly preserved products where no acid is added to the marinade. usually vinegar. Traditionally. and the air movement removes the vapour. it can also be named ‘pickling’ or ‘blood pickling’. without other preservation methods applied after boiling. Usually. and the fish can be immersed in such brines for long periods. Vacuum drying. There are some marinating processes where salt is used for the first cure and a firmer product is obtained. After the freezedrying process. Salted-boiled fish are the products obtained on either boiling fish in a brine. the product rehydrates quickly but does not retain reabsorbed moisture on cooking. Smoking is an ancient process for preserving and flavouring food. where an organic acid. the ice sublimes and the vapour is removed from the fish by vacuum pumps. this ‘brine’ can be collected and placed again over the fish several times.
fish can be smoked by means other than the direct action of smoke. sporeforming bacteria or osmophilic moulds. With the exception of some Asian countries. and filtered to remove tars and particulate matter. 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. by exposure to the wood or sawdust smoke. After the salting step. It is commonly used in place of traditional smoke. which irreversibly combine with the haemoglobin in the tissue to maintain the red colour. including pathogens. A dry or wet salting step is always used in the process before exposure to the wood or sawdust smoke. the salt used in the processes may contain halophilic bacteria. such as injection or dipping in a solution of liquid smoke. Smoked-flavoured fish. Hot-smoking. leaving only elevated levels of carbon monoxide. temperatures below 30 °C are used. the fish is subjected to heat for a sufficient period of time to coagulate protein throughout the fish. or by means of smoke flavouring processes. or liquid smoke.2 Initial Microflora The initial microflora are those of the raw fish from which the products are made. smoking is now used mainly to give the fish a characteristic flavour. Cold-smoking. Smoked fish is prepared by subjecting the fish to the direct action of smoke from wood or wood sawdust burning. The secondary objective is to impart sensory characteristics such as smoke colour and smoke flavour to the product. The process was designed so that the smoke is filtered to remove all smoke particles. The smoke itself may contain spores of moulds. The different spices used in the immersion brines are also a potential source of microorganisms.CURED. 5. the fish is subjected to heat for a period that does not coagulate protein. or added to the brine with other flavourings. or by means of smoke-flavouring processes with liquid smoke. It can be smoked either traditionally. by exposure to the wood or sawdust smoke. Usually. In this process. Liquid smoke is a natural aqueous condensate of wood smoke that has been aged. process temperatures are over 60 °C. In this process. It can be injected into the product. Usually. 95 . which impart the flavour of smoke to fish. However. Also. the sugar used in some products may contain yeasts and moulds as well as spore-forming bacteria. Smoking can be either traditional. and the preservative effect is only slight. Clear smoke. SMOKED AND DRIED FISH of smoke.
about 50º (132.0 g of salt per litre of brine).I).FISH AND SEAFOOD 5. it is impossible to establish standardised processing stages in the curing of fish.0 158. sometimes the salt crystallises onto the surface of the fish.6 g of salt or more per litre of brine) may cause salt burn in this way. smoking or marinating processes.0 .8 211. The dissolution of salt in water is dependent on the temperature. Therefore. a 90º or above brine (237.2 105. all using salting steps.4 184.6 264. a 0º brine saturation means pure water. it causes swelling because the fish absorbs water from the salt solution (1). Different brine strengths are required according to the curing processes and/or the type of product. When working with salting. processes and producers’ private recipes.80º brine (184. or salt in the water phase. For smoking salmon.8 211. A 100º brine solution means that the solution is saturated and no more salt can be dissolved.8 79. the fish is usually salted with a 70 . Salt in the water phase links the two conditions of moisture and saltiness.6 132. it is important to state the temperature. Salt concentration means the percentage of salt in the water contained in the fish. only the most common processes can be described here. it is important to understand the concept of salt concentration.4% salt (Table 5.4 52. is employed.2 237. When a milder brine. When a 100º brine is used.3 Processing and its Effects on the Microflora Owing to the wide variety of products. The strength of the solution is described as degrees of saturation (brineometer degrees). etc. At 18 °C a 100º brine is produced when the solution contains 26. and salt content. % 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.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. TABLE 5.2 g of salt per litre of brine). Salt in the water phase is the amount of salt compared with the total amount of water and salt in the fish. Thus. This should be distinguished from the salt content. which is the percentage of salt in the whole weight of fish that is water plus protein plus fat.
1. can lead to the decomposition of the centre of thick muscles before it reaches the level of salt concentration sufficient to prevent spoilage.35% and 0. It takes about 3 weeks until proper salt penetration and moisture loss are achieved. can be dried further to produce ‘klipfish’ or ‘bacalão’. It is common for the dried salted cod in the Mediterranean countries to be made using solar salts. SMOKED AND DRIED FISH 5. respectively. and such delays. On the other hand. The fish is then stacked in piles of about 2 metres and the water is allowed to drain away.1 Salting Salting of fish for preservation purposes has been used since pre-history. Usually it is followed by other preservation techniques such as drying and/or smoking. respectively (1).0. low in calcium and magnesium. 97 . A ‘green cured’ product is obtained when the water content has been reduced to about two-thirds of the initial amount. The salt used should be of food-grade quality. When pure salt is used.15% calcium and magnesium. the fish is mixed with one part of salt per eight parts fish.15 . having a short section at the end of the tail. when the air humidity is high. Also.1 Dry salting / Kench curing Usually. 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. which consequently inhibits the diffusion of the water from the bulk of the product to the surface. and the salt has penetrated throughout the fish (3). See Figure 5. it tends to lead to a slight yellowing and softening of the product. however. This is because fatty fish turn rancid quickly. The calcium and magnesium impurities.3. For a light salting. Salting can be done in a number of different ways.3.1. for which there is a high demand in Mediterranean countries. 5. ‘Green cured’ split cod with about 60% of the backbone removed. producing characteristic pink patches. when rock salt is used the ‘dun’ patches caused by moulds are prevalent (1). and a ‘putty’ fish product is also obtained (1). the fish is restacked with fresh salt after about a week. and for a heavy salting a ratio of 1:3 salt:fish is used (1). The most common fish used in the northern hemisphere is cod (Gadus morhua). lean white fish is used for this purpose. outgrowth of halophilic bacteria may occur. are recommended to help make the product whiter. addition of levels of 0. and salt can speed this oxidation process. ‘Salt burn’ might occur when the surface of the fish dries out too quickly as a result of the small grain size of the salt. However.1 ppm and 10 ppm. are likely to cause hydroscopicity of the product and. which include calcium and magnesium. and essentially free from iron and copper (3). if very pure sodium chloride is used for curing purposes. This condition is known as ‘putty fish’ (4). but the concentration of copper and iron must be kept under 0.CURED. caught in North Atlantic waters. mainly in tropical conditions.
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. eviscerated and/or split). it appears that water activities much less than 0. the process is mainly for flavouring purposes prior to the use of other preservation processes.3. Fig. The greatest barriers to salt penetration are the skin and scales. when a light brine is used. so that the risk of spoilage is high. Only small fish can be salted whole. Where fish are salted in their whole form. although toxin production would be unlikely (4).1.5.90 are rarely achieved.FISH AND SEAFOOD Dry salting can also be used for small species without the need for opening the fish flat. such as 98 . and the uptake of salt is dependent on the type of fish (fatty or lean) as well as its presentation (whole. Food-poisoning organisms.2 Wet salting or brining As referred to above.1. Processing of dried salted cod 5.
It is then taken out and dried in the sun or in kilns until about 35% moisture in the product is obtained.26 4. Anchovies are the main fish used for these purposes.10 1.65 2. the brine is then replaced over the fish. There are several variations of this process.46 4.96 1.3 Dry / wet salting In this process.66 5.17 4. For example. the water that comes out of the fish forms a brine in which the fish stays immersed for 2 .90 (1).0 4.86 2.0 3. sardines.74 9.04 5.17 5. Histamine formation is of importance when using scombroid and some clupeidean species.85 3.25 5.23 2.3 days. Here.11 3.78) (1).41 4.16 7.56 6.45 3.41 5.76 4. The brining step for smoking purposes also helps the product to develop a desirable glossy appearance.06 4. and a red burgundy colour will develop in the product.73 1.33 5.23 3. Histamineforming bacteria do not generally grow at water activities below 0.76 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. the product obtained is called gaspé-cured (5).98 5.66 5. after being drained and pressed at 465 kg/m2 for 15 hours. but as it is placed in containers. can be cured in a saturated brine solution for 6 days and.84 1. with pressure being applied to the top of the container to help the brine to drain away.95 3. TABLE 5.76 4.09 8.25 5.70 3.26 7.51 3.II. thus.98 6.41 3.26 1. they can be stored at 28 °C. the fish is dry-salted.05 7.49 4.26 3. and can be collected and replaced again over the fish. the liquid that comes out of the fish forms a brine.00 3.76 99 .1. See Table 5.11 4.23 2. SMOKED AND DRIED FISH smoking.5 3.03 2.35 4. The main objective of the brining method of salting fish is to limit air penetration during processing.76 4. Fresh sardines.47 6.23 8. with a shelf life of about 4 weeks (water activity (aw) about 0.88 5.3. a saturated brine is used.60 2.51 5. 5.61 3.30 2.5 4.99 2.09 8.76 4.0 2.44 2. For preservation purposes.03 4.80 5.78 6.54 5. herrings and other fatty fish can be used.19 3. headed and gutted.78 2. but mackerel.61 3.98 6.5 10.26 2.70 6.11 9. if split fish is dry-salted.85 3.64 4.41 6.91 2.5 3.CURED.44 2. The fish are kept immersed in their own brine near saturation point for several days.22 6. limiting the amount of rancidity that can develop. This is why it is the preferred method for fatty fish. the fish enzymes play an important role by maturing or ripening the product.
and packed (3). and even death. they can be cut and cured for 3 days in a solution with 3% white distilled vinegar. underlines the need for careful control (4).2% acetic acid and 2 .30 minutes. salt.4% of salt) for distribution and sale (1). ‘ikan kaju’. Afterwards. only the nonspore-forming organisms are eliminated in the process and. Usually. giving a cooked appearance to the product. usually requiring refrigeration below 10 °C for longer-term storage. flavouring (sugar. There is epidemiological evidence that these products have been the cause of some 100 . However. usually flavoured with onions and hot chillies.2 months. has a shelf life of several years. Herring can also be cured for several days in an 80 to 90º brine with 2. Another salt-boiled Indonesian product. they are packed for consumption within 2 weeks at chill temperature (6). The first process can give a longer shelf life. They will have a salt content >6% (w/w) in the water phase.FISH AND SEAFOOD 5. drying and salt-boiling steps until a hard translucent block is obtained (4). Another variation uses sour cream instead of the second brine step. and lasts for 3 . but it has further smoking. containing 1 kg of salt for every 25 kg of fish. up to 1 . The fish can be soaked in a high concentration of vinegar (about 4 . Once cooled. The Indonesian ‘pindang’ is made by boiling the fish in a saturated salt solution and it is sold as is. The lime juice plays the role of the vinegar. until the flesh is soft.2 Salt-boiling Fish can be boiled after being dry-salted in a watertight container and heated for at least 1 hour. After this. The cooked portions are poured into bowls containing gelatin dissolved in water (10% solution) and sometimes a small amount of vinegar.7%) for as long as 3 weeks. the occasional implication of pindang in cases of foodborne illness. even if the product is hermetically sealed.5% 120grain distilled vinegar. herring or other fatty species are used. Ceviche. A much firmer product is obtained when the fish is salted prior to acidification.3. All these products are ‘semi-preserves’. In a second step. The product is usually consumed raw with the high strength acid solution softening the bones enough for them to be eaten. or can be boiled by being placed in a basket. then packed in barrels with a 70º brine. which is dipped into a boiling brine for 10 . it can be repacked in milder brine (1 .3 Marinating / pickling Fish can be brined in solutions based on organic acid instead of salt. Other preservation techniques can be used for extending the shelf life of these products. and spices) is added to the brine.6 days without refrigeration (1). or a pH <5 and they are stabilised for a limited period. As referred to above. outgrowth of bacterial spores after processing may occur (4). whereas the latter method preserves the fish for only a few days. 5. a Peruvian product.3. Traditional jellied eels are a product where pieces of fish are cooked in boiling water. because of the salt content of the product. placed in a marinade of lime juice. and 6% salt. is made of different varieties of fish.
which are placed in a mixture of salt. sugar and spices. Gravad fish is a Scandinavian product made of salmon fillets. by using either solar heaters or mechanical dryers. SMOKED AND DRIED FISH foodborne diseases related to the presence of biotoxins. still made without prior salting on a large scale. air flows naturally. Matjes are made of fat young herring. and the flesh will break down. Stockfish (dressed.0) (7). some fish species such as moray eel and varieties of dog fish. under a light pressure.3. Shark pectoral fins. South America and the Caribbean. used as an appetiser. 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). after frying. gravad fish.) where no acid is added to the process. The most common method used for fish products is the air-drying process. Fresh dill is placed between the layers of fish. Usually. Some bacteria and enzymes are still active in the presence of acid and salt. Foodborne pathogens. It is important to assure the quality of the raw material.CURED. They are an important dried product. At pH levels below 4. the fish is dried naturally in a relatively cold environment.5. depending on the thickness. In the first case. are generally air-dried at sea. In Scandinavian countries. are likely to survive in these mild-processed products. then mild-cured with special seasonings and packed in barrels with 80 º brine (5). Other products. Dried shrimps are widely used in Asia. mixed with salt. 101 . Dried octopus is also a very common dish in the interior of the country. the fish is dried after salting or brining. etc.4 Air drying This is probably the oldest method of fish preservation. 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. it will last for many years (5). Stockfish is extensively used in Portugal. it being preferable to use frozen salmon or to freeze the product at the end of the process to avoid parasite survival. and a fine product is obtained.There are several products named marinades (matjes. while in the latter case there is a provision for fans or other means of mechanically moving the air (1). a very fine delicacy for Oriental food lovers. split and dried gadoid species) are traditionally made in cold climates. but sometimes it is dried without prior salting. jellyfish. such as squid. marinated products will keep for several months at a temperature of 4 °C. 5. bacterial toxins and parasites (7). gutted. are commonly dried in Asian countries. It takes about 2 days to marinate completely. Italy and Spain (5). In some coastal areas of Portugal. such as Listeria monocytogenes and Clostridium botulinum. the temperature and the amount of pressure applied (5). and the stacks are placed in the refrigerator. Stockfish products have a golden brown colour and are very hard. They are not cooked prior to consumption and they have a limited shelf life even at refrigeration temperatures. Provided the product is kept in dry and fairly cool conditions. are sundried and. sea cucumbers and seaweeds.
the ‘green cured’ cod is rinsed or washed with water and then hung to dry either by the sun and breeze or. tarpaulin. the protein must be set or denatured with low-temperature drying before applying higher temperatures. Owing to restriction of fishing areas for Spanish and Portuguese vessels since the 1970s. The product is dried to a final water content of about 25 to 38% (3). France. more commonly nowadays. Whilst the production of dried fish products is tightly controlled in industrialised countries. Italy and South America. Thus. the quality is more difficult to control.the lower the humidity. the better the moisture transfer. When no more water can be removed from the fish. 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. Spain. by warm air circulating in a drying chamber. the skin will form a hard case (case-hardening). dried salted cod (bacalão) is now mainly produced in Norway. there is an increasing demand from people that have emigrated to industrialised nations for traditional products.g. Iceland and Canada. and it is exported to Portugal. Portuguese and Spanish fisheries cannot satisfy their own countries’ demand for cod. or wooden drying rack to dry in the sun. The fish must be stored in reasonably dry conditions to avoid moisture absorption. botulinum growth might occur in the bulk of the product. neither it. Whilst the low aw of the final product will inhibit microbial growth. and increasing amounts of such products are making their way into the market place. that may have contaminated the product during the drying process. After dry salting. 102 .83 (9)). Dried salted products must have a water activity below 0. botulinum). aureus) or spores (e. Staph. In conclusion. Traditional processing conditions provide ample opportunity for product contamination from a wide range of hazards and sources. which leads to mould growth (aflatoxins. making a water barrier and consequently. Although controls are in place to ensure that products processed in such relatively uncontrolled conditions do not enter the supply chain. but fast enough to avoid spoilage caused by bacterial and enzymic activities (1). spoilage and possibly C. In many such artisanal fisheries the drying is simply conducted by spreading the fish on the beach. It is important to balance the drying rates and quality/safety issues. the product can be held for months at room temperature. skin up.FISH AND SEAFOOD When the fish is sun-dried in tropical conditions. and has the primary objective of creating a product with specific taste and textural characteristics. toxic secondary metabolites of the moulds Aspergillus flavus and Aspergillus parasiticus are not produced at aw <0. if the rate of water removal is too high in comparison with the diffusion rate.75 (1).g. and the water must be changed several times before cooking. However. nor subsequent cooking operations will affect the presence of toxins (e. A highly demanded dried fish product in Mediterranean countries is dried salted cod. C. They must be soaked in water. The amount of moisture transfer from the fish to the air is dependent on the relative humidity . To prevent this. the diffusion of the bulk water to the surface of the fish is blocked.III. typically in stews and soups. some of which are listed in Table 5. the fish must be dried slowly enough to avoid case-hardening.
SMOKED AND DRIED FISH TABLE 5. herring. aureus (toxin) C. Hardwoods are considered better because of the milder flavour they impart to the food. such as Escherichia coli. Staph. It can be used for cold or hot smoking. sprats.salmon.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. Blends of various woods can be formulated to produce sawdust with specific flavours. is mainly a process for flavouring fish rather than a preservation technique. making full description impossible here. Maintaining the temperatures of pyrolysis between 425 and 200 °C.CURED. white mangrove and abura has an inhibitory effect on some microorganisms.5 Smoking Smoking today. Preventing the occurrence of carcinogens during smoking has been an issue for the past several years. aureus and Saccharomyces cerevisiae (10). eel. temperature and humidity can be automatically controlled and the process is faster because the directed air movement draws the smoke through the fish. swordfish. oysters. 5. softwood colours the product quickly. There are many fish species that are used for smoking purposes . botulinum (spores) Pesticides / Chemicals Physical Glass / sand etc. Top-quality fish must always be used. It has been reported that the smoke from mahogany. are some of the ways of reducing the polycyclic aromatic hydrocarbon (PAH) compounds (10). using electrostatic filtration of the smoke. Proper cleaning and preparation prior to smoking and washing of the fresh or frozen fish by water spray or a continuous 103 . clams. tuna. cod roes. The smoking procedures vary widely. 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. However. In the mechanical kiln. Aroma and flavour are a blend of smoke components (3). mussels and many others from tropical countries. mackerel. and the use of smoke generated by superheated steam or liquid smoke. trout. as referred to above. cod.3. haddock.
Also. the following step is salting. All the smoked products. wooden boxes or master cartons (10). Thus. which is important for the appearance of these products (4). Depending on the type of product. or quick-frozen and stored at -18 °C (2).10% of the weight of the fish. Sugar can be added together with the salt. The salting period is dependent on the size of the fish and its presentation (whole. 5. but it will increase towards the end of the process (10). after being smoked need to be chilled immediately to 4 °C before being packed. all cold-smoked products should be cooked before consumption. shrink packaged. Some herring products. Sugar. Thus.or hot-smoked. However.FISH AND SEAFOOD water flow system with chlorinated water (25 . thus allowing the deposit of smoke on the fish surface. Wet salting or brining is preferable to dry-salting when smoking fillets because. tuna and swordfish. pasteurisation) temperature is not achieved in any stage of the process. whether cold. spices. are boiled before smoking (12). as it retains more moisture. skin off or on. smoked cod fillets and finnan haddock are usually cooked before consumption. Brining at 10 15 °C is recommended (10). and the cold smoke is denser. it will reduce the salty flavour. owing to the elution of soluble proteins. thawing must be done slowly for 12 . Drying should be minimal at the start. 104 . but a ‘safety’ (i. The fish can be dry-salted using a weight of salt 5 . usually dry-salted and cold-smoked. they can be vacuum packaged.50 ppm) should always be done just before process initiation (3). an attractive glossy pellicle on the fish fillet surface is achieved after draining. are not cooked prior to consumption. covering the fish surface completely (10). In cold-smoking. it will take up salt twice as fast as fresh fish (10). such as smoked salmon-trout.18 hours at 4 °C. it will double the drying time in the kiln to get the same total weight loss as for dry-salted fillets (11).1 Cold-smoking The major difference between cold. skin on or off). filleted.3. the highly desirable cold-smoked salmon.5. gutted or filleted. temperatures must be kept under 30 °C throughout the process. If wet salting is chosen. Whether the product is prepared headed. Cod roes. If frozen fish is used. permitted colourings or smoke flavours can also be added to the product in the brining step. This difference is crucial for safety issues because the flesh is not cooked. however. low temperatures will slow the uptake of salt. but this concentration will again depend on the type of product and the length of the brining stage. canned or bottled. temperatures and times used in processing coldsmoked fish are very favourable for the proliferation of food-spoilage and foodpoisoning microorganisms. The maintenance of temperatures below 30 °C avoids protein coagulation. When frozen/thawed fish is used. 70 – 80 º brine is commonly used. as well as many other products prepared in the same way. The length of the smoking process for such products is much greater than for the hotsmoked fish.and hot-smoking is the temperature of the process. giving the product the desired appearance and flavour (4). the excess salt is removed with water. or packed in retortable foil pouches.e.
Furthermore. are of concern when smoking fish from tropical waters (13). can be isolated from seafood. thirdly. as. but the proteolytic types A and B. if a 105 .3. Smoked salmon fillets are usually vacuum packed whole or after slicing. but they should be cooled to a temperature of 10 °C within 3 hours.CURED. a drying period at 30 °C. and hotsmoked salmon (kippered) (3). such as Yersinia enterocolitica. refrigerated storage and vacuum packaging still allow its growth (12). gravad salmon (also called gravad lax or gravlax). condensation will occur inside the pack. they are more sensitive to sodium chloride and unlikely to grow in smoked products (13). On the other hand. in order to produce toxin. Afterwards. Pathogenic vibrios. secondly. Smoked products can be also packed under controlled-atmosphere packaging. See Figure 5. stored at 4 °C and packed. there is no point in the process that can fully assure the absence of L. Vacuum packing is recommended only for coldsmoked salmon.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. an initial humidity of 70% seems to be preferable because it is a good compromise between maximum smoke adsorption and minimum casehardening (1). can grow with salt levels of 10% in the water phase. A typical temperature set is as follows: firstly. a final cooking period at 80 °C (12). 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. further cooled to 3 °C within 12 hours and then packed. During smoking. SMOKED AND DRIED FISH During cold-smoking. and their sensitivity to salt (13). typically mesophilic. The exception is vacuum packed fish. botulinum type E the product must have at least 3% salt in the water phase and be stored below 5 °C.5. the products are cooled. a partial cooking period at 50 °C.2. if present. Length of the smoking process at each stage will depend on the species and the type of product. if they are taken from a chill room at 3 °C and packed. Vacuum packing extends the shelf life of the lightly smoked products by suppressing the growth of other bacteria and moulds. Neither the smoking temperature nor the salt content is enough to kill the organism. monocytogenes. The protein must be set or denaturated with lower temperatures before the application of higher temperatures to avoid case-hardening. To avoid growth of non-proteolytic C. botulinum type E present an even longer time than before to grow and produce toxin (14). and the recommended gas mixtures are 60% carbon dioxide and 40% nitrogen (15). which underlines the danger of storage at abuse temperatures of >10 °C (13). If the refrigeration chain is maintained. thus allowing any psychrotrophic C. Although other human pathogens that can grow at refrigeration temperature. This temperature should be maintained during storage and distribution (3). These products should be vacuum packed at room temperature. Hot-smoked products require a longer cooling stage. 5.
5.2. Process of cold-smoked sliced salmon 106 . and moulds will grow on the product (12). it will have a reduced shelf life.FISH AND SEAFOOD product is packed too warm without being cooled to room temperature. Fig.
but seems. white fish about 8 days. a very expensive and fashionable smoked product in the Netherlands. Natural smoke extracts. such as yeast derivatives. Masmeen is usually prepared from bonito tuna by eviscerating and filleting the fish. synthetic smoke flavours. They are generally not cooked prior to consumption. Boiled or steamed clams. Vacuum packaging retards the onset of rancidity when fatty fish is used. are used. moulding it into long thick sections.3 Liquid smoke Smoke-flavoured fish represents a small percentage of the total smoked fish industry output. fatty fish will remain in good condition for 6 days. Katsuobushi. chilled or freeze-thawed. Mackerel. In terms of safety. but mechanical drying is used instead of sun drying. 18). Hon kare bushi is a form of Katsuobushi from which shavings are cut to make aromatic stocks for use in Japanese dishes (24). A type of dried hot smoked tuna called masmeen or masmin is produced in the Lakshadweep islands of the Indian Ocean. 23). 21. The product is often allowed to go mouldy by incubating it in a chamber containing a natural fungal flora. The product is removed from the drying cycle.3. 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). apart from the improved appearance. The composition of commercial liquid smoke can vary widely depending on the method of manufacture. hot-smoked fish can be classified as a ‘heat-treated or pasteurised’ product (7). cooking in brine. are usually 80º brined for 5 . then hot smoking and finally sun drying it. Aspergillus tamarii) and several Penicillium spp. The classical liquid smoke is 107 .3½ hours using the typical temperature set for hot-smoking referred to above (12). Aspergillus oryzae. are usually hot-smoked after being immersed in 80º brine or dry-salted (6). including Eurotium repens and Eurotium rubrum. including ‘umami’ (16. and substances unconnected with smoke but with a smoky flavour or smell. Eels.CURED. The production method is similar to masmeen. (Penicillum cyclopium and Penicillium puterellii) (19). When refrigerated at 3 °C. resulting in a characteristic flavour (20. break down of fats and the moderation of smoke flavour. SMOKED AND DRIED FISH There are several species that are usually hot-smoked and the process can vary markedly.5. 5. and many closely related Aspergilli (Aspergillus ochraceus. they can be stored for 6 months at -30 °C and for even longer when frozen in vacuum packs (12). They will keep well frozen. 17. Hot-smoked products made from white fish will usually keep better than those from fatty fish.6 hours and smoked for 3 . 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). is a boiled smoked fish product that is used in Japanese cooking to impart flavour. Aspergillus ostianus. to have no further advantage over other packing methods for other fish (12). similar to masmeen. 22. Growth of these moulds on the tuna results in the extraction of moisture. rewetted and remoulded to give the desired shape and consistency.
5. smoking. so as to impart an unnatural red colouration to the flesh. 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. When liquid smoke was used.6 to 2. and less than 0. Fresh fish spoilage may be measured by 108 . There are two groups of chemicals of concern in smoke: polycyclic aromatic hydrocarbons (PAHs) and nitrosamines. injecting it into the fish flesh.4 Spoilage Brining. The synergistic effect of liquid smoke with other preservatives could reduce the amount of salt in the product when light diets are necessary whilst still meeting the necessary safety criteria (26). drying. the salt level could effectively be reduced from 4.4 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. According to some recent research.3. both of which are considered potential carcinogens.03 μg/kg of benzopyrene. it has been reported that the liquid smoke-treated product has less pronounced aroma and flavour (25). or applying it as an atomised spray within a modern automatic smoke kiln. Smoke-flavoured fish processed using liquid smoke can be prepared by including liquid smoke in the brine. or smoke extracts in organic solvents. consistency of flavour and colour. 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. established in its guidelines that smoke flavourings added to the food product must assure a level lower than 0. spraying is preferable because it is easiest to control flavour and acceptable with this method (3). Whilst neither the CO or the carboxymyoglobin complex are harmful in themselves. In experiments comparing traditionally smoked fish with liquid smoke. applying it as a dip after brining. Liquid smoke was considered to provide advantages of safety. pickling and marinating fish were originally used to lengthen the short shelf life of fresh fish. The European Scientific Commitee for Food. the result of this process is seen as a manipulation of the natural degradation of tuna colour that hides the true nature of the product quality and therefore has been made illegal in the EU.06 μg/kg of benzoanthracene in the final foodstuff (28). botulinum types A and E spores in several fish species that were stored at 25 °C for 7 and 14 days (25). nutritional neutrality and antioxidant activity (27).FISH AND SEAFOOD smoke condensate that is dissolved in water or oil. 5.8% and still provide protection for 7 days (25). It has been shown that liquid smoke can reduce the formation of PAHs (3). 5.
but.000 cfu/ml of brine may build up. which are found in the salt. which may also be produced to describe the quality attributes and their loss over time for processed fish. and. the fish are cut and splayed to allow greater surface contact with the salt. otherwise. Stronger and longer brining has an effect like that of pickling. Spoilage needs to be carefully defined for each product. or extended refrigerated storage. marination or drying). for some. the fish are first cleaned (known as ‘rousing’) by a salt rub to remove any outer slime. as the fish are added. these bacteria tend to continue to dominate the flora in the liquid. however. and of smoke to add chemical preservatives.. Even saturated brines allow some halophilic organisms to grow. the pickle and dry-salting preservation processes require further processing for significant shelf life increases (e. Micrococcus spp. Most marinated and smoked fish products are first brined or salt-pickled. and high concentrations are usually required to inhibit their growth fully. pickling and dry-salting all introduce bacteria to the fish. the bulk of the microflora comprises Bacillus spp. This is not usually the case for smoked fish as the residual salt concentration is not high enough to inhibit all the non-halophiles. In salts.g. Pickling and/or the anchovy process uses solid salt extraction of fish fluids to form a brine solution (cure). limits the types of microorganism capable of growing. Endogenous marine fish bacteria prefer to grow in the presence of salt (marine concentration approximately 2. the consequent reduction in the pH will help to inhibit spoilage organisms. The use of drying and/or salt to lower the water activity.. For some products. or their products require refrigerated storage. therefore. When heavy brining or salting is important for product shelf life. mainly comprising corynebacteria and Gram-negative bacteria from the fish and water (29). spoilage is due to endogenous microorganisms and enzymes of the fish. solar salts investigated carried the highest load and included halophilic organisms as a result of their production method (open-air evaporation of sea water). contact with the bacteria 109 . a process similar in microbiological effect to that of saturated brining. Often. chemical oxidation. The microflora attached to the fish is relatively unaffected by light brines used before smoking. both of these will affect the spoilage patterns. a very dry product is required to produce 'shelf-stability'. a product may become undesirable for consumption as a result of changes in its sensory characteristics. for others. Some brines are re-used over a period of days and a load of 10. Once these changes are defined. these halophilic organisms may grow and cause spoilage. Brining. Within the limits of safety. the production process will modify the microbial flora present on the product. Upon salt packing. When acid marinating or smoking. and coryneform bacteria. Once placed into brines. SMOKED AND DRIED FISH grading schemes or quality indices. the shelf life may be estimated and investigation made into the causes of the loss of sensory quality. the fish flora are transferred to the brine mix.3%). Usually. rock and manufactured salts did not usually contain halophilic bacteria (29).CURED. also. Salts used for brining have been found to carry a microbial load of between 10 and 1000 organisms per gram of wet salt. contaminant bacteria from other materials used to produce the product.
Basipetospora halophila. Studies conducted on dried and salted fish from Indonesia (31. or after drying if the fish is stored in excessive humidity. it is possible to wash off the infected parts. Aspergillus wentii. gaspé). repens and E. Therefore. The bacteria are osmosensitive and will lyse in water. The condition describes the growth of the halophilic moulds (Wallemia sebi. rubrum are also common isolates from dried fish. like the ‘pink’ organisms. which. (syn. ‘dun’ is not characterised by the production of offensive odours. ‘Sliming’ or ‘putty’ fish is a condition that occurs in poorly or insufficiently salted cod (e. powdery growth. rancidity due to lipolysis. together with a range of Aspergilli. lactic acid bacteria (LAB) and moulds. and Halobacterium spp. yeasts and fungi. may also be found in the salt used in production. 32. halotolerant or halophilic micrococci. Bacteria from these two genera also grow sufficiently in brine and even salt to cause pink discolouration. yeasts. sometimes covering the whole of the fish with a whitish. although it may occur in other salted fish. ‘pink’ and ‘dun’. Only halophilic microbes may grow. the best way of controlling the organisms is to refrigerate the fish. sebi is considered to be the main spoilage agent of dried fish produced in temperate regions (30). At the pre-drying stage.FISH AND SEAFOOD in the salt is made and the flora will develop accordingly in the blood pickle. Eurotium amstelodami.g. though only the latter two species were considered likely to have grown on the fish (32). Aspergillus sydowii. Insufficient salt penetration into the flesh leaves areas where Gram-negative rod-shaped bacteria and micrococci grow and cause a putrefactive spoilage. including A. 33). spore-forming bacteria. Unlike ‘pink’. B. including hydrogen sulphide (26). W. or fat breakdown) caused by enzymes rather than by microbial growth. were also isolated. Sporendonema sebi)). halophilica was isolated 110 . most notably Penicillium citrinum and Penicillium thomii. as little if any growth will occur below 5 °C on salted fish (29). 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). and still produce a reasonable dried product. flavus. niger. it was found in about 50% of samples examined. if not too contaminated. Several Penicillium spp. pisce was the main fungus isolated. as the water activity of saturated brine solutions is low enough to inhibit most bacteria. spoilage of these pickled and dried fish in refrigerated storage tends to be due to chemical changes (textural changes due to proteolysis. A number of spoilage organisms can grow and produce conditions such as ‘sliming’. The metabolism of the organisms will produce off-odours. The microbiology of heavily salted products is dominated by Gram-positive. 34) have shown that P. This condition is often associated with case-hardening. The bacteria form coloured colonies on the surface of salted fish prior to drying.. A. (syn Oospora halophila) and Polypaecilum pisce are two white halophilic moulds that are associated with spoilage of salted fish (31. ‘Pink’ describes a condition associated with surface growth of the pigmented halophilic bacteria Halococcus spp. protein breakdown. E. and Aspergillus penicilloides. Their growth produces noticeable spoilage of the fish appearance but is also controlled by refrigeration.
33. SMOKED AND DRIED FISH infrequently at low levels. where a high internal temperature is reached. In Vietnam. Technical studies demonstrate the ability of many of the xerophiles above to grow at typical water activities seen in dried salted fish (32. Aspergillus clavatus and Aspergillus niger. Vacuum packing limits chemical oxidation. or any smoking process where temperatures in the interior of the fish do not reach >50 °C for a significant period of time. 36). Fish processed by cold-smoking. 111 . it was. Aspergillus spp.747. Pseudomonadaceae. as already stated. whereas a mixture of Enterobacteriaceae and LAB were prevalent in brineinjected salmon (37). Vibrionaceae and Enterobacteriaceae may all play some part in shelf life limitation. Cold-smoked fish will contain microbial flora representative of that of the raw fish. Marine vibrios were the dominant flora in smoked salmon that was drysalted. The preservative effect of the smoke will inhibit some spoilage microorganisms. so the most probable spoilage organisms will be spore-formers or post-smoking contaminants. including A.. Table 5. but this will not prevent the growth of spoilage bacteria. The brining to 3 . which does not create the salty conditions required for spoilage by those halophilic microorganisms. will have a different residual post-smoking flora from 'hot-smoked' fish. Refrigeration is necessary.IV. around the water activity of saturated sodium chloride. and microbial spoilage is more likely.CURED. with a lower aw limit for growth of 0. LAB. It has been reported that the type of salting (dry. The heat during hot-smoking will kill vegetative microorganisms but not all spores. but. Refrigeration of hot-smoked fish will greatly reduce the number of organisms able to grow and will lengthen the shelf life. has been isolated from dried fish (35). botulinum type E.or wet-salting) in the cold-smoked salmon process leads to different microflora growth at the onset of spoilage. For more information on spoilage see Table 5. Reports for vacuum packed cold-smoked salmon have identified more than one organism associated with spoilage. Smoked fish products tend to have a light brine treatment.V gives details of pH ranges and aw minima for the growth of some important spoilage microorganisms and pathogens (38). able to grow on dried salted fish. but often colour changes due to chemical oxidation of the smoked product will limit the shelf life. not the growth of spoilage organisms. so the bacteria spoiling these products will be psychrotrophic or psychrophilic.5% sodium chloride in the water phase will inhibit some bacteria. however. The salt in the water phase must be high enough to inhibit the growth of C. flavus. most marine bacteria are moderately halophilic and are not effectively inhibited by these low salt concentrations.
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. and Halobacterium spp.variable. marinated and smoked fish Spoilage controlled by pH Heat Ant1 Ref2 Process Product Aw Probable spoilage organisms None Fresh fish spoilage organisms Halophilic spore formers and PPC4 Halophilic spore formers and PPC Halophilic organisms: Pink-Halococcus spp. Matjes v3 + ++ v - +++ ++ ++ - v +/- ++ ++ ++/++/- Pindang Bacalao + +++ Gaspé cure Anchovie ++ Anchovies ++ - - - ++ ++ Saturated brined fish Salted then boiled Salted.IV Spoilage of salted. halophila Halophilic spoilage organisms Halophilic spoilage organisms. Histamine producers Halophilic spoilage organisms Spore formers and PPC None Acid-tolerant spoilage organisms Xerophilic moulds Fresh fish spoilage organisms. + to +++ strength of preservative effect 4 Post-process contaminants 112 . . sebi and B.W. boiled. Enterobacteria and PPC Spore formers and PPC Variable Salt marinades Salted then boiled Brined then boiled Salted then dried Canned Fish Gravad.FISH AND SEAFOOD TABLE 5.not used/unchanged. LAB. Dun.
3% sodium chloride in the water phase is enough to inhibit growth of type E for at least 30 days (39). Since neither the smoking nor the drying parts of this curing process are particularly severe. botulinum. the importance of cold-chain integrity must be emphasised. the fermented products being the most dangerous group (7). A number of surveys have shown that fish from temperate waters is mainly contaminated with the psychrotrophic C. However.75 0. the non-proteolytic type E strains can grow in up to 5% sodium chloride.5% of salt in the water phase and great care must be taken to avoid any temperatures >10 °C (13).8 1. salted and pickled products accounted for 10.9 0. SMOKED AND DRIED FISH TABLE 5. respectively. and as botulism is a serious hazard in foods.1 Pathogens: Growth and Survival Clostridium botulinum In the last few decades.5 5.5. The presence of salt in cured products has a great effect on the growth of the bacteria. and C.5 4. the products were heavily salted. 9 and 8 of the outbreaks. In the United States. temperature ≤5 °C and salt ≥3% in the water phase is sufficient to prevent any growth of type E. Examination of 165 outbreaks of botulism caused by fish products showed that smoked.5 0-11 3.61 Organisms Most Pseudomonadaceae C. It has been shown that. botulinum type E Staph.5-8. although the combination of the two factors. under optimal conditions. sodium 113 . botulinum Staph.5-10 4.97 0. but the concentration of salt in smoked salmon or trout (usually varying from 1 4% salt) is not high enough to prevent growth. could not grow and produce toxin.5-10. Before. pressure on the traditional methods of curing. aureus Yeasts Moulds Lactic acid bacteria pH 5. smoked and dried. The concentration required to prevent growth at room temperature can vary from as low as 3. although approximately 10% salt in the water phase is necessary to fully inhibit the mesophilic types A and B strains (14). but the presence of the mesophilic types A and B is of concern in fish from warmer waters (13). botulinum type E. aureus Most spoilage bacteria Most spoilage moulds Halophilic bacteria Xerophilic moulds Aw 0. with this change.5-8. types A and B are not affected by 3 . botulinum type E and production of toxin. when present in the raw material.5% to 5%.0-9.5 5.CURED. popular demand for healthy foods has led to a change in the tastes preferred by the public and. but if the fish products are stored at temperatures <10 °C.97 0.8 0. Sodium nitrite in conjunction with sodium chloride is known for its ability to inhibit C.86 0.V pH ranges and aw minima for growth of microorganisms (38) Organisms Most Pseudomonadaceae C.
as they require more handling. if recontamination occurs and storage conditions are favourable. 5. the organism can survive most of the light curing processes.86 and 10% sodium chloride.4 Listeria monocytogenes The psychrotrophic L.5. The human carrier rate can be as high as 60% of healthy individuals with an average of 25 .4.94 (7).5.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. growth and toxin formation may occur. coli all occur on fish products as a result of contamination from animal/human reservoirs. As with other organisms. Once the product is contaminated. the risk of infection can be eliminated by proper cooking. Usually. The organism is mesophilic with a minimum growth temperature of 7 °C. aureus growth. if it contains sodium nitrite ≥100 ppm.or cold-smoked fish should have at least 3. The degree of concern in coldsmoked salmon is low unless temperature abuse >10 °C occurs (13). they do not grow well in the raw material. but requires higher temperatures for toxin production (>10 °C). Infection by Shigella sonnei has increased in the UK. Thus. An outbreak of Shigella in an Italian cruise ship. the organism may contaminate the product. if the person handling the product is a carrier of Staph. has been reported (42). 5. are particularly susceptible to contamination and. aureus. they can survive lightly cured processes when the sodium chloride content is less than 5% and water activities are higher than 0. if storage temperatures are over 10 °C. its occurrence in a final product is due to contamination during processing by an infected asymptomatic carrier with poor personal hygiene. It is a halotolerant bacterium able to grow at water activities as low as 0.5.30% of the population positive for enterotoxin-producing strains (41). As they are poor competitors. but. monocytogenes is a ubiquitous organism that has been detected in fresh fish skin. related to the consumption of a raw smoked fish meal. 5.2 Staphylococcus aureus This is a ubiquitous organism but its main reservoir and habitat is animals and. 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. but.5.0 . Shigella and E. they will grow rapidly (7). when the bacterial load of the product is lower. the human nose.5% salt in the water phase.0% salt in the water phase (30). Sliced smoked fish products. throat and skin. after processing. and the minimum pH for growth is 4. gills and guts as well as in several lightly preserved fish 114 . However. The FDA established that hot. it should contain not less than 3. more importantly. In foodstuffs.3 Enterobacteriaceae The pathogens Salmonella.
which means that the process must conform to a level of ‘no detectable L. do not survive 1 min at 60 °C. most of them are of little concern to public health. a number of fish products.0. Some LAB. Of microbiological concern in the northern hemisphere are two parasites .45 °C. when an outbreak of nine cases of listeriosis was suspected to have been caused by gravad or cold-smoked trout (47). Until 1997. has also been detected in cod (7).9. from seawater fish.95 (45). which. in the EU Fisheries R&D-FLAIR program. researchers are looking for other ways of preventing the pathogen’s occurrence or growth. monocytogenes pathogenic for only specific segments at risk. a short period of freezing either of the raw material or of 115 . As freezing to -20 °C and maintaining this temperature for at least 24 h will kill all the nematodes. SMOKED AND DRIED FISH products such as cold-smoked fish. Unlike some European countries.6 and water activities below 0.the round worm Anisakis spp. which considered L. Marine LAB is part of the natural flora of chill-stored vacuum packed salmon and cod. As its presence has been reported in lightly preserved fish products whose processing chain cannot fully assure the absence of the organism. Pseudoterranova dicipiens. 5. which means that cooking a fillet 3 cm thick for 10 min at 60 °C will kill any worms present (48). pH 4. a round worm. and in CT95-1207FAIR). cold-smoked fish. such as ‘lightly preserved’ fish products.5. Thus. salt levels of 8 .5 Parasites Despite the presence of parasites in fish being very common. favours growth of psychrotrophic LAB. widely used as starter cultures in food processes. They have complicated life cycles. or nematodes. and the tape worm Diphyllobothrium spp.2. the CT 3162-FAIR program. gravad fish. This includes matjes herring.92 . both being detected in cold-smoked salmon. As they are heat-sensitive. 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. monocytogenes in levels at or above 102 per gram (46). are considered unsafe.8 . lightly salted caviar. the only concern to public health is when eating raw or uncooked fish products. there were no reported outbreaks related to cured fish. which include intermediate hosts. have been shown to have an anti-listerial activity in many food products. and other local traditional products (7).12%. Vacuum packaging (VP) of sliced smoked fish creates a microaerophilic atmosphere. The use of LAB in fish preservation is a topic that has been studied in projects (UP. Its limiting conditions for growth are 0 . marinades (ceviche) (43) and gravad fish (44). Two outbreaks in the United States have been reported to be linked to salmon (10). the USA established ‘zero tolerance’ for the organism. The round worms. monocytogenes’ in 25 g of the finished product. in combination with curing agents and refrigerated storage. from freshwater fish.514.CURED.
<104 104 .<107 ≥107 N/A <100 <20 100 . ‡ On occasions some strains may be pathogenic. texture.<103 N/A 100 . E. coli O157 & other VTEC Vibrio. monocytogenes Staph.<100 20 . monocytogenes) (cfu/g unless stated) <106 106 . not detected in 25 g Campylobacter spp. for example.<100 200 .100 >100 N/A Pathogens Salmonella spp. These foods fall into category 5. smell.VI Guidelines for the microbiological quality of ready-to-eat smoked fish.<104 20 . Acceptability is based on appearance.<104 present in 25 g ≥103 >100 ≥104 ≥104 <103 103 . cereus and Bacillus subtilis# <20 <20** <20 <10 20 .<100 10 . perfringens B. 5. and unpasteurised yoghurt. 48h ± 2h) Indicator organisms‡ Enterobacteriaceae E.<105 ≥105 * Prosecution based solely on high colony counts and/or indicator organisms in the absence of other criteria of unacceptability is unlikely to be successful. (not L. soft cheese. salami. † Guidelines for aerobic colony counts may not apply to certain fermented foods. the organism should be identified # N/A denotes not applicable 116 . and the levels or absence of indicator organisms or pathogens.FISH AND SEAFOOD the final product should therefore be included in the processing as a means of controlling parasites (7). 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.<104 100 . coli (total) Listeria spp.6 Published Microbiological Criteria TABLE 5. If the Bacillus counts exceed 104/g. cholerae Vibrio parahaemolyticus L.<100 20 . aureus C.<100 >104 >100 N/A N/A <20 20 .
Introduction to Fish Technology. 56. USA. 3273.M. Huss H. Torry Advisory Note No. Regenstein C. 254-82 5. 1997. Eds. Wareing P. 6. Smoked. 117 . Assurance of Seafood Quality.H.7 1. Ed. 4. Flick G.. London. New York. salting and smoking). 9. Leatherhead Publishing. New York.W. Birnie K. Flick G.M. Martin R. Horne J. 1993. 2007. Doré I. 8. in Fish Processing Technology. FAO Fisheries Technical Paper. where 30 ppm is permitted Paralytic shellfish poisoning* Amnesic shellfish poisoning * *These criteria are applied to all fish 5. Food spoilage fungi. 1979. Fernandes R. Torry Advisory Note No..E.A. monocytogenes Salmonella spp. 82 (revised). Hall G. in The Seafood Industry. McLay R. 120-38..8 ppm (saxitoxin equivalent) 20 ppm domoic acid. SMOKED AND DRIED FISH TABLE 5.F. 1994. The Smoked and Cured Seafood Guide. Moody M. References Regenstein J. in Micro-Facts: The Working Companion for Food Microbiologists. or presence of toxin 0. coli (ETEC) L. Horner W. Fernandes R. Bannerman A. Chapman & Hall. 1984. aureus * Vibrio cholerae V. 1982. 3. Hot Smoking of Fish. Handling and Processing Eels. Marinades. 334.. Torry Advisory Note No.CURED. Van Nostrand Reinhold. cured and dried fish. 1991. Catching. 1990. Eds. 2. Van Nostrand Reinhold. Preservation of fish by curing (drying. 381-406. 7. Urner Barry Publications Inc.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. Leatherhead... 37(revised).F. 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. parahaemolyticus Vibrio vulnificus C. * Staph. except in the viscera of dungeness crab. Wareing P. botulinum * Limit of tolerance 1x 103 ETEC/g.
C..H.. 1981. 56 (6). 1992. Effect of dried bonito (Katauobushi) and some of its components on GABA-A receptors. in Fisheries Processing. What’s in a name? Are MSG and umami the same? Chemical Senses. Hayashi K. Kinoshita Y. Teeny F. 13. Oxidation of acetophenone by Aspergillus species and their possible contribution to Katsuobushi flavour. 118 . 840-6. 17. 2324-5. Inhibition of Clostridium botulinum types A and E toxin production by liquid smoke and NaCl in hot-process smoke-flavoured fish. Biotechnology and Biochemistry.. Recommendations for the Preparation of Smoked Salmon.. 1995. Ninomiya Y.M. 88. 1980. 45 (10). Jeppesen V.. 51-81.. 1977.K. Aoshima H. Shuto Y.FISH AND SEAFOOD 10. Huss H.E. 5 (revised). 22. 23. Doi M. Food Reviews International. 18. Biotechnology and Biochemistry. Yajima I. Druschitz A. Mitou M... Yokayama S. Degradation and omethylation of phenols among volatile flavour components of dried bonito (Katsuobushi) by Aspergillus species. Doi M. Ogiwara Y. Matsui M. Bioscience. The discovery of umami.E. Eklund M. Torry Advisory Note No. 59 (12). Kanayama T. Kinoshita Y. 22 (revised). and Biochemistry.C.M. Food Chemistry. Embarek P.. Fish Drying and Smoking: Production and Quality. 1995. London. Shiemori Y. 61 (8). Bannerman A.. 1386-7. Bioscience. Control of biological hazards in cold smoked salmon production. Lancaster. Ed. 1051-5. 108 (3). Ninomiya M. Cann D. Perusing Asian soup options.A. Bioscience. 843. Martin A. 935-41.. Dillon R.F. 6 (4). 20. 25. Halpern B.. O-methylation of 2.. Doe P. Yamauchi H.P.. 17 (9). Horne J. Martin A. Microbiological control for fish smoking operations. 958-60. Chapman & Hall. 6-dimethoxy-4-methylphenol by Aspergillus glaucus and their possible contribution to Katsuobushi flavour. Paranjpye R. Lindemann B. Pelroy G. Packing Fish in a Modified Atmosphere. Peterson M. 1990.. Matsui M. Food Control. 1994. 16. 1998.. 19. 18.. 15. Bannerman A. 6 (6).. Cann D.R. Biotechnological Applications.B. 553-72. Botulism and Fishery Products.. 53 (4). 1982.. Food Product Design. Doi M... 2002. 1997. 21. Hosokawa M. Shuto Y. Torry Advisory Note No. Agricultural and Biological Chemistry.. Processing Cod Roes. 14.. 11. Patel T. Shuto Y... 24. Torry Advisory Note No.M. Doi M. 27 (9). 2008.. Biotechnology. Sakakibara H. Torry Advisory Note No. 508.. Journal of Food Protection. Asymmetric reduction of acetophenone by Aspergillus species and their possible contribution to Katsuobushi flavor. Technomic Publishers.W. Flavour constituents of dried bonito (Katsuobushi). 1984. 1989. 2007. 12. 335-40.
M.. Ed.A. in Fish as Food. 1996.. 1997. Hess E. 311-7..D. Jay J. Tour highlights production and uses of smoke-based flavours.H. New species of fungi from Indonesian dried fish.E. Interactions among xerophilic fungi associated with dried salted fish. ceviche. Studies on salt fish. Journal of the Fisheries Research Board of Canada. Shewan J. Diyaolu S. Hocking A.D.V. 36. Pszczola D. 1995. 5. 34. 49 (1). 29. Academic Press. Fungi associated with Indonesian dried fish. London. Food and Drug Administration. 70-2. 119 . 107-11. Maga J.R.M.C. Commission of the European Communities.. 218-25. in Fish and Fishery Products Hazards and Controls Guide. 1986. Press Inc. 29 (2). The control of botulism in hot-smoked trout and mackerel. 742. 41. Hocking A..F. Mycotaxon.. National Academic Press.E. Taylor L. 1991. 98100. 1995. Lancet. Wheeler K.I. Sprouse R. Lucas F.. Sirvas S. 3 (4). Townsend J. Smoked and smoke-flavoured fishery products. Committee on Evaluation of the Safety of Fishery Products. 1996. 27686. in Reports of the Scientific Committee for Food (34th Series). 1971. 351-7.I.A. Cann D. Anggawati A. Letters in Applied Microbiology. 164-9. 1979.K. Borgstrøm G. Antimicrobial properties of smoke. 1994. 1961. Seafood Safety. 28. Frank M.O. Golden D. SMOKED AND DRIED FISH 26..A. Hocking A. 1991. 27.D. 74. Pitt J. Scientific Committee for Food. Importance of autolysis and microbiological activity on quality of cold-smoked salmon. Simini B. Gill T. Blackie. 74 (2)... Incidence of Listeria monocytogenes in an acidified product. Pitt J.. Fuchs R.CURED... in Smoke in Food Processing. Effects of sodium chloride and relative humidity on growth and sporulation of moulds isolated from cured fish. Fungi and Food Spoilage. 2005. Food Technology. An outbreak of Shigella. 43.. Luxembourg. 40. Nahrung. 32.I. 42.A.. Hocking A. CRC.B..M. Smoke flavourings. Springer Science. Hansen L. 1994. 1985.. Fungal flora of South Vietnamese fish and rice. 74. US Food and Drug Administration. Oxford. Ed. Modern Food Microbiology. Food and Drug Administration. 38. Studies on Sporendonema epizoum from ‘dun’ salt fish. 348. Røntved S. Ed.. Rockville. 31. 35. 37. Journal of Food Technology. 123-9. Cox J. V. 22.. Food Microbiology. 38 (3). Huss H. Loessner M. 14. Boca Raton. 181-8. Ahmed F.T.1988. New York. 1941.S. 1993. Wheeler K.F. 39. 12. Ed. The microbiology of sea-water fish.D. Journal of Applied Bacteriology. Washington DC. 197-208. Journal of Tropical Medicine and Hygiene. Adebajo L.J. 88-90. 33. Pitt J.Y. CEC. Food Research International. Institute of Medicine. 30.
Washington D. Canadian Journal of Microbiology. An outbreak of listeriosis suspected to have been caused by rainbow trout. Mentzing L-O. 80. The effect of NaCl on the upper temperature limit for growth and enterotoxin synthesis by Staphylococcus aureus.M. salted. Danielsson-Tham M-L.. Communicable Disease and Public Health. Davies A. 48...L. Pelaez C.. 45.. 3. Richards J. 2001. US FDA Centre for Food Safety & Applied Nutrition. Incidence and growth of psychrotrophic Clostridium botulinum in food.. Food and Drug Administration. Appendix 5: FDA and EPA Guidance Levels. Amsterdam. Danielsson-Tham M. 1995. 1992.. Inhibition of Clostridium botulinum types A and E toxin formation by sodium nitrite and sodium chloride in hot-process (smoked) salmon.. Cured. 46.. Eds.C.S. 1980. 45 (9).. 507-10. Eklund M. Harrison M. Garren D. Peterson M.. Liston J. Microbiological guidelines for some ready-to-eat foods sampled at the point of sale.fda..FISH AND SEAFOOD 44. 1991. Ed. 120 . 1982. 1994.. 26 (4)..M. Tham W.A... Prevalence of Listeria monocytogenes and other Listeria spp. Beltran A.. 197-209. Huss H.H. Journal of Clinical Microbiology.. Hughes A. US FDA Centre for Food Safety & Applied Nutrition.V. 2000. Zeitschrift für Lebensmittel-Untersuchung und –Forschung. Pelroy G.. and smoked fish establishments good manufacturing practices..html Gilbert R...A. Elsevier Science Publishers B.cfsan.. Paranjpye R. Donovan T. 863-6. in Quality Assurance in the Fish Industry. Moral A. 37 (1).C. 5-7. Acta Veterinaria Scandinavia. 232-6. Unnerstad H. 163-7. Hurst A. Torry Advisory Note No. 35 (11) 2904-7. 49-59.W. 5.N. Jacobsen M.J. Hobbs G. Huang Y..J.8 Further Reading Gibson D. Pathogenic microorganisms of importance in seafood. Little C. Keeping quality of vacuum packed smoked sardine fillets: microbiological aspects. Ribeiro C. in smoked and "gravad" fish. 1980. 55 (4).. Susuki E. de Louvois J.R. 188 (3). 1996. Ericsson H. Loncarevic S. Clostridium botulinum and its importance in fishery products. Nye K. 22.gov/~comm/haccp4x5. 1989. Journal of the Association of Food and Drug Officials. Journal of Food Protection. 135-85. Round Worms in Fish. Journal of Food Protection. Cann D. 5 (1).. 13-8. Fletcher R. 58 (8). Tham W.. Bolton F. Growth and production of toxin of Clostridium botulinum type E in rainbow trout under various storage conditions.W. Roberts D.E. Advances in Food Research.. Food Control. Loncarevic S.D. in Fish and Fishery Products Hazards and Controls Guide. 1997. Eklow A.. 47. Wooten R. 1976. Persson I.. 833-41. Association of Food and Drug Officials.A. http://www. Gibbs P.
309-18. Jemmi T. 1994. Pelroy G. temperature.. Eklund M. 844-6.S. 61 (12). Ben Embarek P. in a salmon slaughterhouse and smoked salmon processing plant. 21 (3). and salt concentration on growth and survival of Vibrio parahaemolyticus. 1994. Paranjpye R. 58 (5).. 1974.R.N.R. Bayliss A. Peterson M.C. Hitchins A.CURED. Nicolaides L. Roberts T. Interacting effects of pH. Rørvik L.A. 14. 1994.. Hudson J. Tran T. Occurrence of Listeria monocytogenes in freshwater farms and fishsmoking plants. Combined effects of water activity.. Journal of Food Safety.178-80.. 309-16. 4389-95. 394-6. Beuchat L.. Journal of Food Protection.. Journal of Food Protection. 1075-80. 11. 293301..J. Elsevier. Davies A. Marine toxins of microbial origin. Beuchat L. 1992. International Journal of Food Microbiology. Yndestad M. 1973. and temperature on the growth of Vibrio parahaemolyticus.E. Letters in Applied Microbiology. Fuchs R.R. Liston J. International Journal of Food Microbiology. detection and growth of Listeria monocytogenes in seafoods: a review. Histamine production by psychrophilic flora. 42.. Caugant D. Dillon R. 25 (5).. 121 . 25.. 1734. Slade A. Efficacy of high sodium chloride concentrations for the destruction of Listeria monocytogenes. Ratnam S. 1988. 502-8. Lashbrook L. 1995. van Spreckens K. SMOKED AND DRIED FISH Sutherland J. 217-36. 94-8. 27 (6). Eds. Fate of Aeromonas and Yersinia on modified-atmosphere packaged (MAP) cod and trout.P. Poysky F. Applied Microbiology. Poindexter C. Patel T. Letters in Applied Microbiology. 1994. Taylor S. International Journal of Food Microbiology. 1987.A. Contamination pattern of Listeria monocytogenes and other Listeria spp. 1995..A. 866-70.. Presence. Kramer D.W.. 55 (11).. Predictive modelling of growth of Staphylococcus aureus: the effects of temperature. 19.T.. Competition of food bacteria with Listeria monocytogenes during enrichment culture. 1992. Food Technology. Prevalence of Listeria in smoked fish.. New York. Keusch A. 19-27. Applied Microbiology. Incidence and sources of Listeria monocytogenes in cold-smoked fishery products and processing plants. Letters in Applied Microbiology. Dallas H. Incidence of Listeria in hot and cold-smoked fish. 1991.. 1995. in Seafood Quality Determination. Food Microbiology. solute. 11. pH and sodium chloride.K. 23 (1).
koji. Sugar can also be added to the Indonesian fish sauce bakasang and the Thai product nam-budu. 123 . the most important of all fermented fish products. Those to which only salt is added tend to be mainly autolytic.1.I. Essentially. there are numerous variants of some common themes and a host of local names used to describe them. in economic terms. producing the fish sauces and pastes. they are exported to European.1 Fish/salt products such as the fish pastes and sauces tend to contain relatively high levels of salt. though some are produced elsewhere. 6. The principal fish sauces and pastes of south-east Asia and their local names are listed in Table 6. Martin Adams Faculty of Health and Medical Sciences University of Surrey Guildford Surrey GU2 7XH United Kingdom 6. the term ‘fermented’ describes a spectrum of processes that ranges from the largely autolytic degradation of fish protein. they can be divided into two categories: fish/salt products and fish/salt/carbohydrate products. FERMENTED FISH Prof. whereas in those where a carbohydrate source such as rice or sugar is also added. to those processes in which the activity of lactic acid bacteria (LAB) plays an important part. added during preparation. Fermented fish products are largely confined to east and south-east Asia. North American and other markets and are. typically in the range 15 . Carbohydrates are added to some fish sauces during their preparation. the best-known of the fish sauces (gyoshoyu) of Japan is shottsuru. The fish sauces are produced on the largest scale.6. lactic fermentation can play a significant role. It is produced from the sandfish Arctoscopus japonicus and has the rice-based enzyme preparation. For example.25% and are used mainly as a condiment. As with most traditional products that are still produced principally on a cottage industry or domestic scale.1 Definitions As applied to fish products. The relative importance of these two activities depends on the formulation of the product.
6. may be direct descendants. and the more widely known Worcestershire sauce a very distant relative. 124 . to products analogous to other lactic fermented foods such as salami. This is well exemplified by some of the fish/salt products of Thailand described in Table 6.2 Fish/salt/carbohydrate products range from those that resemble the fish sauces and pastes in the sense that extensive autolysis has occurred. These have long since fallen from use. Fermented fish products are of very minor importance outside Asia.FISH AND SEAFOOD TABLE 6. In Europe.II.I Fish sauces and pastes of south-east Asia Country Sauce Amber/brown liquid. fish sauces known as liquamen or garum were ubiquitous condiments for the Romans. where the bacterial production of lactic acid is a major feature. made from fish livers in Greece. 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. 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.1.III. and pissala. salty taste. a variety of products can be produced within a single country. who had adopted them from the Greeks. A selection of Asian products is presented in Table 6. made from whole fish in France. although garos.
pla-chao.FERMENTED FISH TABLE 6. funa-suchi. cencalok burong-isda. 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.III Fish/salt/carbohydrate products of south-east and east Asia Country Japan Kampuchea Korea Laos Products I-sushi. pla-som. ayu-sushi. e. tai-suchi phaak. crustaceans Variety of fish Variety of brackish or marine fish 1:4 3 .4 months Condiment and main dish Whole 1:3 18 months Condiment Whole 1:3 After 3 . e.3 months Main dish Tai-pla Bowels 1:3 6 .g. pa-khem. mam-pa-kor. shrimps.12 months 10% raw sugar added. burong-bangus pla-ra. som-pa-keng pekasam. som-fak/ som-fug 125 Malaysia Philippines Thailand . burong-ayungi. som-pa. pa-chao.II Fermented fish/salt products common in Thailand Name Type of fish Form Approx. crustaceans Mackerel eviscerated fish Variety of fish Whole 1:4 Condiment Pla-thu-khem Whole 1:3 2 . the mixture is then boiled and bottled - Condiment Nam-khoei Shrimps. mam-chao. burong-dalag. mam-seeing sikhae som-kay-pa-eun.8 months Main dish TABLE 6.g.
Penaeus indicua Many of the fish used in fermentation are uneviscerated. Table 6. and their microflora tends to reflect their local environment more than that of marine species. in Sweden. for example. relatively mild-tasting product. however. and old descriptions of their production often refer to the addition of whey. made from herring. A modern account of their production. some fermented fish products remain popular. For example. freshwater fish are the usual raw material.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. paddy ponds beside rice fields used for collecting fish when the field has dried up.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. 6. which has been increasing in popularity over the last 50 years. 126 . which employ lower levels of salt than Asian products. Lactic fermented fish products are often associated with inland areas such as the Central Luzon region of the Philippines and the north-east of Thailand. states that sugar is often included with the salt. in a survey of fermented fish products in north-east Thailand. These are often described as fish/salt products. in Norway and surstrømming. it was found that several different sources were used for the fish . TABLE 6. rakefisk or surfisk. the most popular varieties of which are rakørret. There are also more heavily fermented products. malt or flour (1). Norwegian gravlaks. and a large local freshwater reservoir (2). and counts ranging from below 104 /g to in excess of 107 have been reported.IV) and in the way in which they are handled prior to processing means that the initial microbial levels are far from uniform. Here. is a traditional. This could lead to quite marked variations since the same fish can be obtained from a variety of sources. The diversity of fish used in fermented products (see. or buried salmon.FISH AND SEAFOOD In Northern Europe.the flooded rice fields. and the initial microflora is likely to be considerably larger in these cases. fermented trout. especially in fish/salt products.
3. and the anaerobic conditions that prevail in the bulk of the material. These preparations contain a mixed microflora of moulds and yeasts. probably reflecting their ability as spore-formers to survive rather than any capacity to multiply under the prevailing conditions (5). where the total counts dropped from above 107 cfu/g to below 103 cfu/g after 14 days and below 102 after 40 days. after one month. however. either directly or as the partially saccharified rice they produce. This. Bacillus spp. traditional starter culture preparations such as koji (Japan). Hansenula. Endomycopsis and Saccharomyces .1 – 6.4. In another microbiological study of four fish sauces. Similar results were seen with patis. More recently. which included nam-pla and patis. Studies have shown that the number of bacteria declines rapidly during the production of the Thai fish sauce nam pla and that. Other halophilic species have been identified from fish/salt products. angkak (Philippines and Thailand) and look-pang (Thailand) are used. and studies 127 . 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. in many cases. 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.74 or below. Generally. the Philippine fish sauce. sufficient to saturate the water phase present and produce an aw of 0. It has also been shown that a perfectly acceptable patis could be produced from fish that had been sterilised with ionising radiation (6). however. its contribution is likely to be insignificant compared with that of the fish microflora in terms of both spoilage potential and public health concerns. suggesting that halophilic bacteria do play an important role in the production of fish sauce (8). work with nam-pla in Thailand has isolated Halobacterium and Halococcus spp. In some cases. mean that microorganisms have been generally held to play little or no role in the process that follows. 6. It also demonstrated their significant proteolytic activity during the first month of fermentation.3 6.FERMENTED FISH Where other ingredients are used. in numbers reaching 108 /ml after 3 weeks of fermentation but declining thereafter. principally species of the genera Rhizopus. were found to be the predominant isolates. they can contribute to the initial microflora. When this is a simple carbohydrate source such as sugar. 4). 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. Mucor. the product contained about 500 cfu/g comprising mainly Micrococcus and Bacillus spp.organisms that play an important part in the production process but can also contribute to the ultimate spoilage of the product. Similarly microbial inactivation by irradiation has not been found to adversely affect the properties of Korean fermented squid (7). (3.
Patis (Condiment) 128 . Belacan (Condiment) Fig.2.12).1. 6.FISH AND SEAFOOD have demonstrated the production of halophilic proteinases from fish sauce isolates (9 . 6. Fig.
4. 6.6. Myulchijeot (Staple food) The extensive proteolysis and liquefaction that occur during production are. Ika-Shiokara (side dish) Fig.FERMENTED FISH Fig. This is more rapid when whole fish are used since the head and viscera contain higher concentrations of proteolytic enzymes than other tissues. To produce semi-solid fish pastes as opposed to fish sauces. however. 14).3. In fish sauce production. the 129 . thought to be largely the result of autolytic breakdown of the fish tissues. eviscerated fish are sometimes used to slow the autolytic process. and muscle cathepsins have been shown to be inhibited at relatively low salt concentrations (13.
6 – 6. particularly glutamate. Some examples of products and their formulation are presented in Table 6. which is eventually separated from the residual fish tissues.FISH AND SEAFOOD high salt level used also assists in the osmotic extraction of a supernatant rich in amino acids.5. Their main feature is that the provision of a source of carbohydrate allows a lactic acid bacterial fermentation to occur.8. 6. snack) 130 .2 Fish/salt/carbohydrate products of Asia This category encompasses an even greater diversity of products than the fish sauces and pastes.V and Figures 6. volatile fatty acids and nucleotides.5). filtered and bottled. for example. In contrast to free-swimming fish. Hoi-Malangpu-Dong (side dish.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. 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. 6. Fig.
5-1. N.12 months Shelf life Use 1 . N. NE 10 . north-east.7 days 5 .4 are also produced using shrimps. C. N S 5 . Fig. side dish) 131 . Ang-kak: red mould rice obtained by fermenting rice with Monascus purpureus.3 years Condiment and main dish 2-3 months 3 weeks 2 weeks 2 weeks 6 .5:2-3:1 Fish:salt:boiled rice: ang-kak 3:1:3:0.20 days 3 Pla-som 4 Pla-jom 5 Som-fak 6 Pla-paengdaeng NE. C NE.03 Preparation time 6 . S. 3:1:1-3 Fish:salt:boiled rice: garlic 10:2:1:0. C NE. NE Formulation Fish (whole or pieces): salt: roasted rice.12 months Main dish 2 Pla-jao C.V Fish/salt carbohydrate products of Thailand Name 1 Pla-ra Region C.12 days 3 . N. In these cases.25-1 Fish:salt:roasted rice: garlic 10:1:3:1 Fish (minced):salt:rice: garlic 10:0. central.2-4 Fish (pieces): salt:khaomark (fermented rice). the prefix 'pla' is replaced by 'kung'.6. Kung-Chom (staple food.FERMENTED FISH TABLE 6. north.6.10 days 5 days Main dish Main dish Main dish Condiment and main dish Note: Products 1 . 3:1:0. south of Thailand. NE.
It also indicates that.. 2.10%.2. the salt is of far greater importance in determining the product’s long-term stability. 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. Generally. then this will take longer to develop than if lower salt levels are used.VI). Phillipines Journal of Food Science and Technology.1 The use of higher salt levels results in a longer production phase but a better keeping quality product. of the two factors contributing to shelf life .salt and lactic acid. giving a reduced rate of acid generation.FISH AND SEAFOOD The extent to which lactic fermentation occurs depends largely on the salt content. their growth and metabolism are nonetheless slowed. (from Arroyo et al. 106) Fig. If acidity is a desired sensory property of the product.3.6. The LAB isolated from these products are those with more marked salt tolerance but.7. these products are salted at lower levels than the fish sauces and pastes. but there is considerable variation in this. typically in the range 2 . Red burong isda 132 . A survey of fish/salt/carbohydrate products in Thailand (15) noted that there were two empirical rules governing formulation:6. while they are able to grow at high salt levels. This is readily explained by the susceptibility of LAB to elevated salt concentrations.
9.2 4.0 Shelf life 3 weeks 6 .8 .5. 2.5 .FERMENTED FISH (from Arroyo et al.9 . 133 .5.8 pH 4.1 .9 4.5 .17.8 2.0 . 106) Fig.3 5.6.9 4.7 .2 7. White balao-balao TABLE 6.5 3.2 4.1 4.6 220.127.116.11 years 2 weeks 2 weeks Source: A Concise Handbook of Indigenous Fermented Foods in the ASCA Countries (16).0 . Philippines Journal of Food Science and Technology.18.104.22.168 months 1 .6.4..VI Shelf lives reported for fish salt carbohydrate products Product Pla-som Pla-paeng-daeng Pla-ra Pla-chao Pla-chom Som-fak Salt 2.8 .4.0 .4 .3 years 1 .8.3 .
TABLE 6. although the final population of LAB may be similar. 19). also serves as a valuable source of the more readily fermented carbohydrate.4.VII).3.1 . Where they are used.FISH AND SEAFOOD 6. 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 . of course. Although the ability to ferment starch is not widespread in the LAB. When added as boiled or roasted rice alone. For example. being similar (18. also lower the unit cost of the product and this is probably a significant factor in many cases. 21).5) despite other factors. Garlic. The effect of this will be to decrease the product’s final pH and increase the amount of total acidity. In several products. such as salt content. Rice is the most common carbohydrate source used.0 . the rice may be added in a partially saccharified form such as kao-mark (Thailand).3. a survey of burong-isda in the Philippines reported consistently lower pH values for red burong-isda where ang-kak was used (pH 3. 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. they increase the amount of soluble sugars produced and thus the range of LAB that can grow. This results in a more active lactic fermentation and a lower pH. efficient lactic acid production will depend upon the presence of amylolytic LAB or saccharification by other microorganisms or enzymes that may be present. a saccharifying agent such as koji (Japan) or ang-kak (the Philippines) may be added separately (Table 6. a common ingredient in these products. alternatively.VII Fish/salt/carbohydate products in which saccharified rice or a saccharifying starter culture is used Product red burong-isda i-sushi. inulin.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. 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).9) compared with the white product (pH 4.2. Addition of carbohydrate will.
will not reliably exclude air. There is little published information on the factors that limit the shelf life of the low-salt products. A combination of irradiation and chill storage has been shown to prevent both over-acidification and fungal growth in som-fak. Generally.5 in the first 2 .8 . One study found that any effective measure to exclude air from a fermenting fish/carbohydrate mix prevented yeast growth (23). markedly extending the shelf life (25). and there is a marked regional variation in the preferred type. around 12%. is something of an exception since millet is used along with garlic as the carbohydrate source.3 weeks at around 20 °C. such as wrapping in banana leaves.2 at the end of fermentation. the salt levels in the latter are sufficient to rule out microbiological spoilage in all but the most exceptional of cases.FERMENTED FISH Sikhae.2. lead to an increase in pH.5:9:3. Excessive growth and acid production by the LAB could render products too sour for acceptability. The Thai product pla-ra exemplifies these basic rules as well as the difficulties associated with attempts to classify such products. reporting the product’s pH to range from 4. the pH drops from 6. thereafter declining to about 4. The growth of yeasts and moulds could. Results of a product survey were consistent with this.3 days. In a successful fermentation. the Korean fermented fish product.5. 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). Cleaned pieces of eviscerated fish flesh are salted with 6% salt overnight before mixing with cooked millet.17.7 to below 4. Clearly. This suggests that use of oxygen-impermeable wrapping films could extend the shelf life of such products. 135 .7 up to 6. but in most cases it is probable that yeast and mould growth would be an important cause of spoilage. 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. Traditional packing techniques used with lactic fermented fish products. 6. which is sufficient to prevent lactic fermentation by all but the most halotolerant LAB. but they are likely to behave similarly to lactic fermented products produced from other commodities. It seems that a lactic fermentation takes place in many types of pla-ra but is not significant in others. Any loss of acceptability must arise from chemical or physical changes occurring over prolonged storage. allowing other spoilage organisms to grow. which ranged from 7. the level of salt employed is on the high side for fish/salt/carbohydrate products. in turn.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). 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).9%. The mixture is allowed to ferment for 2 . red pepper powder and garlic in the ratio fish:millet:pepper:garlic of 80:7.
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. Because of the severity of the illness it causes. which accounted for 88% of outbreaks in the period 1952 . The combinations of pH and salt in many fermented products are likely to preclude growth of the organism completely and. During fermentation.8 and at salt levels as high 10%. under conditions inimical to growth and high ambient storage temperatures.87. and failure to achieve a sufficient combination of inhibitory factors could lead to their growth or survival. 6. V. The Scandinavian fermented fish product rakefisk was responsible for 16 (84%) of the outbreaks of botulism recorded in Norway in the period 1961 90. Their safety therefore is critically dependent on the quality of the raw materials used and the inhibitory effects of the fermentation process.5 Pathogens: Growth and Survival Many fermented fish products are stored at ambient temperatures. parahaemolyticus has been reported growing down to a pH of 4. and the most frequently implicated product is the fish-salt-carbohydrate fermented product Izushi. outbreaks of botulism are the only well documented instances of illness associated with fermented fish products. As in Norway. however. but we are unaware of any reports of this happening. typically 30 °C. viable numbers will probably decline quite rapidly.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. In Japan. the pH decreases below 5 and safe processing is recognised to depend on proper salt levels and temperature control until the pH drops. Several measures have been suggested for reducing the risk of botulism. The bacterial pathogens most commonly associated with the living environment of fish. fish products are the most common source of botulism. 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. The epidemiological evidence points to greater awareness of the importance of these factors since outbreaks have declined over recent years (27). are Vibrio parahaemolyticus and Clostridium botulinum. although these limits apply when all other factors are optimal (26). fish can potentially be contaminated with almost any foodborne pathogen as a result of improper handling and storage. outbreaks of 136 . Improper storage of fish prior to processing could result in the production of toxins capable of persisting through to the product. and consumed without cooking. In the fish sauces and pastes. The fish/salt/carbohydrate products depend on the combination of salt and pH/lactic acid for their safety. There is little in the way of documented information associating fermented fish products with foodborne illness. After capture.
Dhanamitta S. Novus Press. botulinum and the production of toxin. Lutefisk. 105-10. and salmon heads and eggs. Greenland and Alaska. With this particular organism. affecting an estimated 40 million people. Foodborne trematode infections are emerging as a major public health problem. Thailand. Where these products are consumed without final cooking. They are packed into containers such as buckets or pits in the ground and left at ambient temperature to putrefy. viverrini were a strong risk indicator and that consumption of fermented fish products was an independent risk factor in the contraction of cholangiocarcinoma (31). Infection with the trematode Opisthorchis viverrini is associated with cholangiocarcinoma. Liston J. Kasemsarn B. Many of these products are described as fermented. 1989. beaver tail and paw. probably reflecting greater awareness of the critical factors that determine the product’s safety (27). An investigation of survival of the fish parasitic nematode Anisakis in marinated herring has given some indication of what these studies might find. they could act as an important vehicle of transmission. particularly in south-east Asia and the western Pacific regions (30).. 6. The more widespread current use of non-traditional containers such as glass or plastic jars is thought to be one contributing factor (28). botulinum. Saisithi P. A case-control study conducted in northeast Thailand showed that elevated levels of antibodies against O.. which is generally consumed raw (29). Baseline Study (1987-1988). Freshwater fish are the most important intermediate reservoir hosts of foodborne trematodes. References Riddervold A. One study with the trematode Haplorchis taichui. giving an alkaline fermentation. although the absence of any source of fermentable carbohydrate means that the process is putrefactive. 1966. Fermented Fish Products Study (Thailand) Phase 1. Dollar A. Microbiology and chemistry of fermented fish.FERMENTED FISH botulism have become less frequent in recent years. Journal of Food Science. 1990.6 1. Botulism has also been associated with a number of traditional foods of the peoples of northern Canada. and warm weather can result in rapid growth of C. Salting levels are often insufficiently low to prevent growth of C. salt content was found to have the greatest impact on the viability of larvae. 3. 31.. 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). Rakefisk and Herring in the Norwegian Tradition. Raw materials subject to this process include seal flipper. while increasing the acetic acid content of the formulation had little or no effect (32). Puwastien P. 2..O. whale meat. which will persist into the product. though 27% survival was still recorded (33). Institute of Nutrition. Mahidol University. Oslo. and these are often used in the production of fermented fish products. walrus nose. 137 . showed that inclusion of fermented fish in the Thai dish lab-pla improved inactivation of the metacercariae. Yhoung-Aree J..M.
SK33 isolated from fish sauce fermentation.. Ahn H.V..R. Pakdeeto A. 70... 6338. Aslam Z. 15. 124-30. Adams M.M. Tanasupawat S. 43.. International Journal of Systematic and Evolutionary Microbiology.. 63. Velankar N.. Crisan E. 57. 138 . Thawai C. Suntinanalert P. New York....R. from fermented shrimp paste in Thailand.J..T. A Concise Handbook of Indigenous Fermented Foods in the ASCA Countries. 11.. Backhoff P. 185-92. Reddi P. 6.T.K. Journal of Scientific and Industrial Research (India). Ono H. Dhamcharee B.. 14.H.W. Yongsawatdigul J. Kyong-Haeng L. Takano M. 2008. 3649. Kaneko Y. Liston J.FISH AND SEAFOOD 4. 8. 12..R. Sands A.. Kudo T. SR5-3 isolated from fish sauce: purification and characterisation. 5. Hull R. Biotechnology and Biochemistry. 2006.. 16A. Some chemical changes in fish silage. Applied Microbiology.. Orejana F... isolated from jeotgal. Fukuda H. 1985.. 3.. International Journal of Systematic and Evolutionary Microbiology. 1976. 643-7. Nov. 80. Journal of Food Technology.A. 1395-401. 13. 1979. Cooke R. 25. 1975. 57. 1957..M. Production and characterisation of NaCl-activated proteinases from Virgibacillus spp. Process Biochemistry. 11. Journal of Fermentation and Bioengineering... Namwong S. Halophiles in Thai fish sauce (Nam pla). Itoh T. Hiraga K. Microflora of four fermented fish sauces. 1995.. Im W. Moonmangmee S. Tanasupawat S. 2007. 1986. Rodriguez-Valera F. 106-8.K. 381-8. 934-9. 29. 7.. Protein hydrolysate from fish. Tsunemi M. Phil.. 17. Bioscience. Lim J. 16. microbiological and chemical properties of salted and fermented squid. Olympia M. Sinsuwan S. 141-2. Journal of Food Science. Effects of gamma radiation on sensory qualities. Saono S.. Dong-Ho K. a traditional Korean fermented seafood... Yasir M. Fermented fish products of south east Asia. Byun M. Rattagool P. Agents of proteolysis and its inhibition in patis (fish sauce). 17-30.. 2007. 61-73. Constantinides S. Government of Australia.D. Chung Y. Journal of Food Science and Technology. Lee S. A halophilic serine proteinase from Halobacillus spp.H. 2000. 1991.. Dymsza H. 10. Rodtong S. Ed. Yook H. Nov. 1972. Salinococcus jeotgali spp. 353-63. Lentibacillus kapialis spp. Tropical Science. Catheptic activity of fish muscle.. Journal of Food Protection. in General and Applied Aspects of Halophilic Microorganisms.S. 37. 9. Canberra.. Characterisation of starch-hydrolysing bacteria isolated from a fermented fish and rice food “burong isda” and its amylolytic enzyme. Plenum Press. Jae-Hun K.. Takada K. Oda K. Thongthai C...
.. 2002. 1983. New York. 21. Kozaki M. Two outbreaks of botulism associated with fermented salmon roe – British Columbia. Volume 5: Microbiological Specifications of Food Pathogens.H. 29. Yeast-flora in red burong-isda a fermented fish food from the Philippines. ASEAN Food Journal. 307-14.. 3. Characterisation of lactic acid bacteria isolated from a Thai low-salt fermented fish product and the role of garlic as substrate for fermentation. Martin J.. 1983. 1996. Traditional fermented fish products with special reference to Thai products.. Genotypic and phenotypic characterisation of garlic-fermenting lactic acid bacteria isolated from som-fak.H. 20.W. The influence of oxygen accessibility on the growth of yeast in fish/rice fermentation. World Health Organisation Technical Report Series.D. 22.. Dodds K.. Caldo G. 28. Huss H..L.. 19. Wedell-Neergaard C. 24. Sakai H. 30. Journal of Agricultural Science (Tokyo).. Sakai H.H. 138-44. Marcel Dekker. 27. 28.W. Control of foodborne trematode infections. 2002. 28.. Fermentation and spoilage of som fak.. Food Chemistry.H. Benjakul S. 426-35.B. Journal of Applied Microbiology. a Thai low-salt fermented fish product. Ben Embarek P. in Microorganisms in Foods... in Clostridium botulinum: Ecology and Control in Foods. Paludan-Müller C. Riebroy S. 3-10. International Journal of Food Microbiology. Journal of Agricultural Science (Tokyo). Journal of Food Science and Technology. 103. in the Philippines. Erikson U.. 1990. Hauschild A. 1987. Dodds K. Dawar M.A. 69-104. 1999. Eds. a Thai low-salt fish product. 92. 219-29. Vibrio parahaemolyticus. Wainwright R. Fung C. Owens J. burong-isda.B. Gram L. Avurhi J. 46. Tropical Science. WHO Study Group.. 27. 26. Blackie. 23. in Clostridium botulinum: Ecology and Control in Foods.. 274-86. Eds. Effect of irradiation on properties and storage stability of Som-fug produced from bigeye snapper.. Visessanguan W. 1998. The fermented fish food. International Commission on Microbiological Specifications for Foods..H. 1-4. Saisithi P..L. 38. New York.. 28 (6).W.M.A. Valyasevi R. 305-22. Hauschild A. Paludan-Müller C.. 1992. Canada Communicable Disease Report. Gram L. 139 . International Commission on Microbiological Specifications for Foods. Huss H.. Epidemiology of human foodborne botulism. 1995. Moody L. Marcel Dekker. 1815. Hazards from northern native foods.FERMENTED FISH 18. Gram L.H.D. Ed. 2007. Huss H. Hauschild A. Tanaka M. Ostergaard A..K. 104-6. Rustad T. London. Isaac-Renton J.. 25.. Kozaki M. 105-12.. Patrick D. 1992. Caldo G. Yamprayoon J.
. 2005.J. Ruamsuk A. Miwa M. Jedpiyawongse A. 29. 110-1... Fermented flesh foods. Uchida K. Hanai S.. 36. Aspen Publishers Inc. Sriplung H. 1995.. Survival of Haplorchis taichui metacercariae in lab-pla.... 854-60. 6. 33. Horst K. northeast Thailand. Honjo S. Chuboon S. Wongsawad C..H.. 661-70. Kittiwatanachot P. Bloemsma B.R. Adams M. Todoroki T.R. Gaithersburg. 2001. Deerasamee S. 15998. 117. International Journal of Cancer. Srivatanakul P..R. 23. 2005..7 Further Reading Adams M.... 140 . Progress in Industrial Microbiology. International Journal of Food Science & Technology. Kikukawa H. Sripa B. Genetic and environmental determinants of risk for cholangiosarcoma via Opisthorchis viverrini in a densely infested area in Nakhon Phanom. 1986. Nithikathkul C.. Roepstorff A.FISH AND SEAFOOD 31. Fermentation and Food Safety. Southeast Asian Journal of Tropical Medicine and Public Health.. Nout M. 32. Survival of Anisakis larvae in marinated herring fillets. Huss H. Thai traditional food preparation..
M. the export of exotic seafood that may contain such toxins can lead to unexpected public health issues.1 Introduction Shellfish and finfish toxins continue to be a threat to public health and local fisheries around the world. a number of new toxins and toxin families have been identified. Another book dealing with the clinical effects of a variety of poisons and toxins. Increasingly sophisticated instrumentation (e. several scientific reports have described their efficacy and potential. The manual on harmful marine algae and their products. FISH AND SHELLFISH TOXINS Jeffrey L. and is often necessary in light of the complex toxin profiles that can occur in shellfish tissue. physiology and detection of marine and freshwater toxins has also appeared (8). In the last decade seafood monitoring has intensified and detection methods have continued to develop in selectivity and sensitivity. known and unknown. Nevertheless. While some of these toxins only occur in seafood from a restricted geographical region.3) and some practical applications for detecting shellfish toxins have been reported (4. Liquid Chromatography/ Mass Spectrometry/ Mass Spectrometry -LC/MS/MS) is employed to identify and confirm the presence of known toxins.. produced by the Intergovernmental Oceanographic Commission (IOC) of UNESCO (United Nations Educational. Although most of these antibody-based test kits have yet to receive international regulatory approval. and a second updated edition dealing with the pharmacology. Another important development is the introduction of sensitive and selective antibody-based detection methods for a number of toxins. C. 5). most notably the azaspiracids and the cyclic imines as well as some additional polyethers.g. more traditional methods such as the mouse bioassay continue to be used in many countries to warn of the presence of toxins. Ph.7. Wright. FCIC UNC Wilmington Center for Marine Science Marvin Moss Lane Wilmington NC 28409 United States of America 7.C.. It remains to be seen if this approach gains momentum in the future. Scientific and Cultural Organisation) has been updated (7). Since the original chapter in this series was published (6). An emerging technology is the use of immunosensors (1 . including the well known marine toxins 141 .D.
142 .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. Gymnodinium catenatum. In addition. even crustaceans such as lobsters. 7. temperate. and sub-tropical waters and habitats (15).2. An account of this meeting detailing recommendations and scientific advice on marine biotoxins has been published (10). A brief report on the status of rapid detection methods for phycotoxins in the European Union (EU) has appeared (11). and a tropical species Pyrodinium bahamense (16). and a review on the risk assessment of marine toxins has also been produced (14). PSPs are watersoluble and heat stable so cooking has little or no effect on PSP toxicity.FISH AND SEAFOOD has been produced (9). Around twenty PSP toxins are known.g. Saxitoxin Group instead of the Paralytic Shellfish Poisons). all of which contain a tetrahydropurine skeleton (Figure 7. Saxitoxin (STX) and neosaxitoxin (neoSTX) are the parent members of the group and are the most toxic. dealing with the detection methods and regulatory requirements for many marine toxins has been published (12). a great variety of shellfish from all latitudes. The Ncarbamoyl sulfates are readily hydrolysed to STX. neoSTX and Gonyautoxins (GTX) 1-4. 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. There is a recommendation from a joint FAO (Food and Agriculture Organisation of the United Nations)/IOC/WHO (World Health Organisation) report that marine biotoxins be classified according to their structural chemistry group rather than the symptoms they induce (e. and another. can become contaminated with these toxins. with intoxications occurring in many countries even today (15). All PSPs degrade rapidly in alkaline conditions. 7. some are the result of biotransformation processes by the filter-feeding host. The remarkable frequency of neurotoxins from marine dinoflagellates has been noted (13).1). Consequently.1 Origins and distribution The PSP toxins are produced by several dinoflagellate species belonging to the genera Alexandrium. a number of useful and comprehensive reviews and book chapters have appeared dealing with various topics related to individual toxin groups and these are referred to in each appropriate section.
However. Human symptoms begin with tingling and numbness of the lips. tongue. Cooking does not destroy these compounds and although some of the toxins may be extracted into the cooking water. and fingers within 5 . Structure of saxitoxin and neosaxitoxin and their sulphated derivatives 7.2 Toxicology PSP toxins affect the central nervous system of mammals. 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.30 minutes of consumption. 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).1. the less potent C toxins are readily hydrolysed by acid to the more potent GTX toxins. 143 . this is never sufficient to render the seafood safe to consume.FISH AND SHELLFISH TOXINS STX GTX2 GTX3 GTX5 (B1) C1 C2 NEO GTX1 GTX4 GTX6 (B2) C3 C4 R1 H H H H H H OH OH OH OH OH OH R2 H H OSO3 H H OSO3 H H OSO3 H H OSO3 - R3 H OSO3 H H OSO3 H H OSO3 H H OSO3 H R4 H H H SO3 SO3 SO3 H H H SO3 SO3 SO3 - Fig. and humans seem to be particularly susceptible. 7.2. leading to paralysis within a few hours and in extreme cases death by asphyxiation due to respiratory paralysis.
7. including Japan. Alternative HPLC methods have subsequently reduced this to a single HPLC run. 7. An antibody-based kit that can detect STX in shellfish is commercially available (RIDASCREEN®). While the chemical detection methods are most promising for the future.1 Origins and distribution Unlike many other marine toxins there is no temporal distribution of TTX episodes. 15. TTX most commonly occurs in many species of puffer fish. and over ten analogs are known (22. Early post-column methods were somewhat time-consuming and required three separate HPLC runs to complete the analysis. 17). There continues to be a significant effort applied to the development of alternative chemical and biochemical detection methods for these toxins. and a second rapid dipstick kit (MIST Alert™) is available for the qualitative detection of positive and negative samples (19). 24) (Figure 7. After some development and various trials. Thailand. and has also been evaluated by a number of EU laboratories. The mouse bioassay has been in place for decades and the method has been standardised. 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. One approach has been to take advantage of the ready conversion of these toxins to fluorescent derivatives. and islands of the South Pacific. and more standards of individual PSP toxins are required to validate the complete profile to toxins that could occur in contaminated shellfish tissue. as well as other parts. 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 Tetrodotoxin Tetrodotoxin (TTX) possesses a unique chemical structure and was the first marine toxin to be chemically characterised. Taiwan.2). or conversion before HPLC separation (pre-column derivatisation) and these have recently been thoroughly reviewed (12. 23. specifically 144 . Two approaches have been taken here. Since its discovery.FISH AND SEAFOOD 7. This kit was found effective in all the samples tested in Europe (20) and was applied recently in a survey of Scottish shellfish (21).3 Detection methods The safe limit set by most countries for PSP toxins is 800 µg/kg (0. and while simple and relatively inexpensive to operate. other derivatives have been reported.3. a pre-column method has received Association of Analytical Communities (AOAC) approval (18). China.2.8 µg/g) of shellfish or seafood tissue. Most intoxication events have occurred in South East Asia. However it remains as the monitoring method of choice in many countries. a criticism has been that such methods are time consuming. namely oxidative conversion to fluorescent derivatives after High Performance Liquid Chromatography (HPLC) analysis (post-column derivatisation).
25. Since then.3. gonads and roe (24).2. this has expanded to include bacteria within the Alteromonas and Pseudomonas genera. Typically this has been accomplished using HPLC followed by post 145 . The onset of symptoms usually occurs rapidly. TTX acts on the central nervous system by binding extracellularly to site 1 of voltage-gated sodium channels. where puffer fish of the genus Fugu are a delicacy. and in mild cases results in distal muscle weakness. it is served in speciality restaurants with qualified chefs to properly prepare the fish. and ingestion of larger amounts leads to cardiovascular effects including bradycardia. Fig.3. However TTX has been found to occur in a remarkable variety of genetically unrelated species including gastropods. nausea and vomiting. respiratory failure and coma. 26). 23. and once again the mouse bioassay has been used to detect the presence of the toxin. A review of TTX detection methods has been published (12.3 Detection methods TTX can be extracted from suspect tissue using dilute acid conditions. preventing depolarisation and propagation of action potentials in nerve cells. and even newts. 7. Structure of tetrodotoxin 7. raising speculation that TTX is produced by a bacterial source. and blocks diffusion of sodium through the sodium channel. Severe poisoning causes paralysis and respiratory failure. 27). This was indeed confirmed and a number of marine Vibrios have been shown to be capable of TTX production (24. 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.FISH AND SHELLFISH TOXINS in the liver. and more recently by an actinomycete Norcardiopsis dassonvillei (26).2 Toxicology Like the PSP toxins. All of the observed toxicity is secondary to the action potential blockade.2 mg of purified toxin can be lethal. 7. the blue-ringed octopus. molluscs. An oral dose of 1 . horseshoe crabs. In Japan. hypotension.
and while promising. Since the original episode of human poisoning following consumption of DA-contaminated shellfish in Canada in 1987 (28). need further development (24).4 Amnesic Shellfish Poisoning (ASP) Toxins The parent compound of this group is domoic acid (DA). Structure of domoic acid and three naturally occurring photoisomers 7. but it is known that such congeners are less toxic than DA itself (30).1 Origins and distribution The cosmopolitan diatom Pseudonitzschia multiseries was identified as the culprit organism in the 1987 poisoning event (31). 7. However this method has been criticised for being complex. but all of them are considerably less toxic than DA itself (29). Fig. but 146 . 7. Currently 12 isomers of DA have been reported (see Figure 7. both the toxin and the diatoms capable of producing it have been found to be distributed worldwide. Immunoassay methods have also been investigated.3 for examples).3. Surveys by a number of groups have identified over a dozen species of Pseudonitzschia capable of producing DA. These isomers arise as a result of isomerisation and location of the double bonds in the side chain of the molecule. 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).FISH AND SEAFOOD column oxidation to the fluorescent derivatives.4.
seizures. 7. Recently. and in locations around New Zealand (32. but any impairment of the blood brain barrier results in additional risk. herbivorous fish become toxic after ingesting strains of Pseudonitzschia spp. cormorants and pelicans (34). Four deaths were attributed to the Canadian incident and years later a few seriously affected individuals still display chronic neurological deficits. 33). Levels of DA in the other strains are generally <1 pg/cell. The effects of DA may be augmented by the excitotoxic effects of glutamate and aspartate in the sample. For example. 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. has been reported as a medium-level producer of DA (33). and Pseudonitzschia seriata) are considered to be high producers of DA (>10 pg/cell). disorientation and coma. as does impaired renal function which increases serum concentration and residence time of the toxin. and review of the toxicologic pathology of DA has been published (36). Penetration of the blood brain barrier is poor in healthy subjects. 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.2 Toxicology DA is an acidic amino acid and potent neurotoxin unaffected by cooking or steaming. which has been well documented in studies along the west coast of the US.39). Another factor is the movement of the toxin through the food chain.. scallops. and this has led to ASP toxicity in birds. and clams. southern and western portions of Ireland. 147 . multiseries.10% of the administered dose. Indeed.4. found in the Pacific around Vietnam. the Philippines. 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.FISH AND SHELLFISH TOXINS only three strains (P. The original 1987 event was followed some years later by a more widespread episode along the western coast of the US and today toxin-producing strains of Pseudonitzschia have been found in the Gulf coast of the US and along the western shores of the US and Canada. the Mediterranean. Okinawa and more northerly parts of Japan. Victims display a variety of symptoms including headache. including persistent short term memory deficits (36 . nausea. and sea animals such as sea lions and even whales (35). Pseudonitzschia australis. To a great extent. and is mainly distributed in the blood. The oral absorption of DA is around 5 . 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. another diatom species Nitzschia navisvaringica.
are amenable to laboratory culture and consequently are used in many experiments involving studies of DSP toxins. An EnzymeLinked ImmunoSorbent Assay (ELISA) kit (Biosense) for DA has been approved by the AOAC. but just over 15 years ago a third toxin. In the context of this chapter. 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). Since then. DTX-2 has been found to be extremely widespread throughout many European maritime countries and is often the major DSP toxin present. 41). DSP toxins are produced by dinoflagellates of the genera Prorocentrum and Dinophysis (47. and this is typically the method of choice. the heterotrophic dinoflagellates Dinophysis spp. and dinophysistoxin-2 (DTX-2) (Figure 7. Provided the concentrations in shellfish are high enough (>50 µg/g shellfish tissue). The benthic Prorocentrum spp. 49).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.FISH AND SEAFOOD 7.4). DTX-2 was discovered in contaminated Irish shellfish (46).30 ng/g) with a rapid clean-up step using a strong anion exchange resin. australis. in Monterey Bay. though this method is most commonly used to analyse plankton and seawater (42). 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. Since their first characterisation it was believed that the only causative agents of this group were OA and DTX-1 (45). and these methods have been reviewed (40..5 Diarrhetic Shellfish Poisoning (DSP) Toxins The active agents responsible for DSP incidents are a group of polyethers that include okadaic acid (OA). shellfish tissue can be efficiently extracted with aqueous methanol (1:1). 48. However.4. and consequently were often overlooked as the real source of the toxins. Fortunately DA is conveniently detected by a variety of LC-linked analytical methods and this is the recommended approach for DA monitoring in EU countries. To further complicate matters. 7. Dinophysis spp. produce a variety of other 148 . dinophysistoxin-1 (DTX-1).. Of additional interest is the use of molecular probes originally developed to detect the presence of cells of potentially toxic Pseudonitzschia spp. DA can be detected by the mouse bioassay but the regulatory limit for shellfish is set below this level at 20 µg/g (20 mg/kg) shellfish tissue. The sensitivity of the LC/UV (ultra violet) method (detection limit ~ 1 µg/g) can be improved (20 . Alternatively. 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 an immunosensor method has also been reported (4). Pseudonitzschia fraudulenta and Pseudonitzschia delicatissima were found to be less effective. are the more potent source of toxins – often very low cells/ml of seawater can result in human poisoning. California. although in the latter study the probes for Pseudonitzschia pungens.
Today over ten Dinophysis spp.. Structure of okadaic acid and the two related DSP toxins 7. Fig. which include the pectenotoxins and the yessotoxins. 7. 149 . are recognised as DSP toxin producers. and include Dinophysis acuminata. and Prorocentrum belizeanum.FISH AND SHELLFISH TOXINS lipophilic compounds that are toxic in the mouse bioassay. 49). and DTX-6) all of which are rapidly converted to OA when the plankton cells are ruptured (48). but do not have the same mechanism of action as DSP toxins nor induce the same symptoms. Dinophysis fortii. such as Prorocentrum lima. 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. In this case Dinophysis was suspected as the culprit organism. 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 acuta. DTX-5. While these esters are not toxic in vitro. Norway and Japan were likely due to DSP toxins produced by a Dinophysis spp. In comparison. This was probably not the first DSP episode. are also recognised as a threat to human health. they can be converted to the parent toxins by nonspecific esterases or lipases either in the producing organism or in the host shellfish. DSP toxins produced by benthic Prorocentrum spp. analysis of Prorocentrum cultures have uncovered an additional series of diol esters of OA and sulphated diesters (called DTX-4. and Dinophysis norvegica among others (47. Lately these compounds. and anecdotal evidence now suggests that several seafood poisoning episodes during the 1960s and 1970s in the Netherlands.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. The parent acids are the only DSP congeners to be found in shellfish tissue.5.4. Dinophysis caudata.
various European countries. DSP episodes have been reported worldwide and are included in many monitoring programs (47). All the DSP toxins have around a thousandfold greater affinity for PP2A than PP1. The onset of symptoms can occur within 30 minutes of consuming contaminated shellfish and generally last about 2 .3 days.2 Toxicology The DSP toxins are potent inhibitors of the serine/threonine phosphatases PP1 and PP2A. Immunoassays have been developed for OA detection but an inherent problem of these methods has been the low cross reactivity with DTX-1. and while this provides critical seafood safety information. which can lead to an underestimation of the total DSP content in a sample. Since the late 1970s. Due to the lipophilic nature of the toxins. New Zealand and Chile. DSP episodes have been reported from Japan. and abdominal pain. and a detection limit of 100 µg/kg has been reported (12). even with the minimal of clean up steps (48). 7. some careful clean-up steps are required to avoid such matrix interferences and to simplify data interpretation (48). although a comparative study of in vitro phosphatase inhibition with mouse bioassay data has shown that OA is about twice as potent as DTX-2 (51). nausea.160 µg OA equivalents/kg of whole shellfish tissue has been implemented in some countries. DSP-Check is a commercially 150 . A regulatory level of 80 . 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.5.3 Detection methods Once again the mouse or rat bioassay is relied upon to detect DSP toxins in contaminated seafood. vomiting. In recent years.FISH AND SEAFOOD Perhaps as a consequence of these multiple sources of DSP toxins. Human symptoms include diarrhoea. The 7-O acyl derivatives (DTX-3) of OA that can form in shellfish also show poor cross reactivity and this further exacerbates the problem. or indeed on the presence of other associated LSTs that can occur with DSP toxins and which result in a positive mouse bioassay result (vide infra). and DTX-2.5. it was agreed that the established mouse bioassay (52) with an observation time of 24 hours is currently the preferred method for the detection of the acute toxicity of acetone soluble DSP toxins. it does not provide information on the DSP toxin profile. This regulatory level includes the possibility of LSPs in the DSP fraction being tested. 7. both critical enzymes involved in many key metabolic pathways in mammalian cells (50) and consequently affect a host of other biological processes in a cell including tumor promotion. but the method is complex and matrix effects as well as interferences with other lipophilic compounds such as the pectenotoxins compromise the method. 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.
Some representative examples of both groups of toxins are shown in Figures 7. and another rapid antibodybased test kit for DSP toxins has also been reported (53). 58).5. As a result these compounds are now being referred to as lipophilic shellfish toxins (LSTs) (49). In yet another approach.6. Within this group. with good agreement with the mouse bioassay and LC–based methods (54. Structure of the principal pectenotoxins and one of the seco-acid derivatives 151 . Indeed.5 and 7. with a claimed detection limit of 20 µg/kg. 7. they are toxic in the mouse bioassay and can result in false positives and hence are included here for completeness.6 Lipophilic Shellfish Toxins (LST) Lipophilic extraction of toxic shellfish for DSP toxins often results in the simultaneous extraction of other lipophilic polyethers such as the pectenotoxins (PTXs) and yessotoxins (YTXs). over 20 PTXs are known (56) and more than 30 YTXs have been reported (57. for many years these compounds were included in the DSP toxin complex. 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. Fig. However. 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.FISH AND SHELLFISH TOXINS available ELISA test kit that can detect both OA and DTX-1 with comparable sensitivity. 55). 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. and the latter group are usually found to be responsible for any reported shellfish toxicity (59). 7.
Structure of the two parent yessotoxins. As is typical for many algal species. eastern Canada. YTX and homo-YTX 7.6.6. 7. While 152 .2 Toxicology PTXs show varied toxicity in the mouse bioassay (59). 7. 57. New Zealand and South America (49.1 Origins and distribution It is now well recognised that PTXs are co-produced with DSP toxins by various strains of the dinoflagellate Dinophysis and hence the distribution of both families of toxins in shellfish is similar (49).FISH AND SEAFOOD Fig. 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. 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). The sources of YTXs have now been identified as the dinoflagellates Protoceratium reticulatum and Lingulodinium polyedrum.6. many parts of Europe and the Mediterranean. but nevertheless the toxins are extremely widespread and have been found in various shellfish in Norway and Scandinavia. the toxin content and profile may vary among different strains. which may co-occur in blooms with other organisms. 58). Japan.
6. some of which are found as glycosides and sulfates.7 Neurotoxin Shellfish Poisoning (NSP) Toxins NSP episodes are caused by a family of fused polyether compounds known as the brevetoxins (63. The crude extract may be used directly in the mouse bioassay. 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. Their toxicology and mechanism of action remain uncertain. it may be compromised by other biological materials in the matrix. but less so following oral dosing (61). the PTXs are susceptible to bioconversion in shellfish tissue and consequently it is often desirable to determine the profile of toxins present. but a fluorescent derivative has been prepared for others. Some of the PTXs are potent cytotoxins in vitro (60). For many years it was believed that 153 . Acetone extracts of shellfish tissue are dried and re-suspended in 60% aqueous methanol before partitioning against dichloromethane. the PTXs show considerably less toxicity when administered orally. a regulatory level of 1 mg/kg shellfish tissue has been set.7). but various congeners are acutely toxic in the mouse bioassay following IP injection. chemical analysis is best carried out by LC/MS and several approaches have been described (57. 64). 58). Because of the number and complexity of the YTX group. though its cell effects may be linked to modulation of calcium channels (62).3 Detection methods Typically these toxins are co-extracted with DSP toxins following acetone or aqueous methanol extraction of the shellfish tissue. There are two skeletal types known as PbTx-1 (or brevetoxin A) and PbTx-2 (brevetoxin B) (Figure 7. Estimates of acute toxicity of the parent compound following IP injection vary considerably. There are no clear cases of human illness resulting from ingestion of PTXs (59). Although there is no evidence that yessotoxins are responsible for human illness. and this is believed to be due to hydrolysis of PTX-2 to the seco acid derivative. However. 7. The EU has recommended an extraction and solvent partition process to separate the polar YTX compounds from the more lipophilic PTX and DSP compounds. which aids detection by LC/UV. which is further compounded by differences in the experimental design. 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). While LC/UV detection is possible for PTXs. 7.FISH AND SHELLFISH TOXINS demonstrating considerable toxicity following IP injection. Many YTX toxins contain a conjugated diene system. The YTX group has not been reported to cause human illness.
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”. Fibrocapsa japonica. namely Chatonella marina. PbTx-1 and PbTx-2. Many of these species are associated with massive fish kills. brevetoxins are found in mussels. was chemically characterised as PbTx-2 (67). 7. and Heretosigma akashiwo.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 off the coast of New Zealand (65). the same toxins have been reported to be produced by three genera of Raphidophytes.S. verruculosa off the mid Atlantic coast of the U. and cockles (65. These are biotransformation products of principally PbTx-2 by addition of certain amino acids to the conjugated aldehyde function (66). Interestingly. which are found in Japanese waters (66). 154 . In the Gulf of Mexico. oysters.FISH AND SEAFOOD NSP toxins only occurred in the Gulf of Mexico and along the western coast of Florida. and recently the major toxin associated with an ichthyotoxic unialgal bloom of Chatonella cf. but in 1992/1993 they were identified as the culprits in a human poisoning event in New Zealand (65. whelks. They have also been associated with fish kills along the eastern mid-Atlantic coast of the US. Structure of the two parent brevetoxins.7. Fig. 66). and one of the brevetoxin metabolites BTX-B2 7. 66).7.
In a comprehensive multi laboratory evaluation of five different detection methods as possible replacements for the mouse bioassay. 66. the competitive ELISA method correlated most closely with the mouse bioassay results.7. beginning with herbivores that feed on benthic dinoflagellates associated with reefs and which are 155 . pre-treatment or clean-up of the shellfish sample is essential in order to obtain reproducible results. which can result in a cascade of associated events in a cell (66). and in severe cases seizure. and it has also been used in studies to quantify occupational and recreational exposure to aerosolised brevetoxin exposure (73). 7. Ciguatera toxins (CTXs) are extremely potent and occur in finfish rather than shellfish. with hot spots in the coral reef areas of the Caribbean and French Polynesia. shellfish homogenates. this family of toxins has proven to be a challenge both to identify and monitor. 7. Exposure to brevetoxins can also occur through inhalation of brevetoxin aerosols resulting in irritation of the upper respiratory tract (65. and mammalian body fluids without pretreatment or dilution (72). While the mouse bioassay continues to be used. 69). The brevetoxin metabolites are reported to be less toxic than the parent compound (70). nausea.3 Detection methods Three basic approaches for the detection of NSP toxins have been recently reviewed (71). 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. Ingestion of brevetoxins results in a variety of symptoms including parathesia. The mean time to onset of symptoms is around 3 hours. with a mean duration of symptoms of around 1 day. considerable progress has been made in the development of LC-MS methods to routinely monitor for brevetoxins in shellfish.2 Toxicology Brevetoxins bind to site 5 of voltage-gated sodium channels. The mouse bioassay is used as the primary detection method. The human health implications following exposure to NSPs have been reviewed (69).7. but this method is now used routinely in New Zealand and the US to monitor for the presence of brevetoxins (68). and diarrhoea.FISH AND SHELLFISH TOXINS although recently it has been shown that herbivorous fish can store brevetoxins in their flesh (68). vertigo. A second generation competitive ELISA assay has been developed that has successfully quantified brevetoxins in a variety of biological and environmental matrices including seawater. malaise.8 Ciguatoxins Known for centuries and originally referred to as ciguatera by Spanish explorers in the Indo-Pacific ocean and Caribbean. In this application. 7. Outbreaks of ciguatera poisoning are confined to tropical and sub-tropical regions of the ocean.
and only a few structures have been identified. These include species belonging to the genera Ostreopsis and Coolia. a group of closely related polyethers that are considered the source of ciguatoxins. A detailed comprehensive review of ciguatoxins has recently been published (74). Distribution through the food chain begins with 156 . Structure of the Pacific variant of ciguatera toxin (P-CTX-1) and the Caribbean variant (C-CTX-1) 7. 7.1 Origins and distribution It is recognised that several species of benthic dinoflagellates can produce CTXs and the gambiertoxins. lipid soluble polyethers.8.FISH AND SEAFOOD transferred through the food chain to the larger carnivores. but more are suspected to exist. The diversity of benthic phytoplankton that exist on reefs further complicates the picture.8. green and brown seaweeds and also occur in sediments and coral debris. Indeed even the chemistry of CTXs from the Pacific differs subtly from those found in the Caribbean (Figure 7. CTXs are large. most likely a consequence of biotransformation of the parent toxins as they pass up the food chain. and other genera of neighbouring reef dinoflagellates can further contribute to an extremely complex profile of toxins in finfish. complex. Fig. Culprit species producing CTXs may simultaneously produce unrelated toxins such as maitotoxin. Such microalgae contribute to the primary production of tropical and sub-tropical reefs and live in epiphytic association with bushy red. although the dinoflagellate Gambierdiscus toxicus is considered to be the major source of these toxins.8).
2 Toxicology Ciguatoxins (and maitotoxin which is often co-produced with CTXs) are extremely poisonous. The pharmacological effects following CTX exposure are numerous and include severe gastrointestinal distress and neurological symptoms. A good knowledge and careful observation of the symptoms in the bioassay (Minimal 157 . 7. and there are suggestions that such incidents are increasing and spreading worldwide (74). and the sensation of loose teeth (77). 7. and moray eels. and it is estimated that ingestion of as little as 0.1 µg of ciguatoxin can result in human illness (74. These include paresthesias (numbness and tingling of the extremities). and inducing membrane depolarisation resulting in prolonged symptoms indicative of damage to nerve tissue. Patients have reported symptoms lasting weeks and even months. though the mouse bioassay continues to be relied upon as the most common method of detecting CTXs. and temperature sensation reversal. myalgia. the absence of a strong chromophore means that simple detection methods based on UV or fluorescence-based detection methods are ineffective.3 Detection methods As a consequence of their potency. Even when suspect samples are available. all of which can be linked to the effect of CTX on excitable membranes. 75. Nevertheless CTX poisoning continues to be a threat to local and visiting populations. arthralgia. Differences in potency among the CTXs correlate directly with their binding affinity for the sodium channel.8. The onset of symptoms can occur rapidly within 24 hours. 75). but fish toxicity can vary widely depending on where the fish are caught and how long they have been feeding in that area.8. Another symptom that can develop over time is extreme fatigue. only trace amounts of CTXs can cause illness following consumption of contaminated fish. 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. which in turn are consumed by carnivores such as snappers. there are neither standards nor an official testing program. sharks. Like the brevetoxins. 78). pruritus. Other symptoms include a metallic taste. although not all CTXs are equipotent. groupers. they bind to voltage-dependant sodium channels in the cell membrane causing them to open. Globally. which can return following the consumption of alcohol and certain foods such as oily fish (74. the neurological symptoms can persist for days.FISH AND SHELLFISH TOXINS grazing by herbivorous fish including the sturgeon fish and parrotfish. and although the gastronomical problems may dissipate within a day or so.76). which can be confused with chronic fatigue syndrome. Local island populations attempt to avoid poisoning by catching only smaller fish. barracudas. where cold objects feel hot and vice versa. weeks or even months (74 . and in fact CTX poisoning is often only identified after consumption of contaminated fish.
A sodium channel-specific assay using mouse neuroblastoma cells has been described although this approach has still to be developed in a routine. Since the original discovery and identification of the toxin. Immunoassays may be an alternative detection method (79). which features characteristic fragment ions corresponding to sequential losses of three molecules of water.FISH AND SEAFOOD Lethal Dose (MLD) 0. The majority of these have been proposed based on LC/MS analytical data. 158 . This is then subjected to LCMS/MS analysis. high-throughput screen (74).5) have been fully characterised by Nuclear Magnetic Resonance (NMR) and MS analysis. only a few laboratories are able to undertake such analyses. Chemical detection methods are severely compromised by the lack of standards.20%) present in lesser amounts (Figure 7. following consumption of contaminated mussels Mytelius edulis (82). and another dealing with the chemistry and detection methods has been published (85). at C-3 and C-23).25 µg/kg) can provide useful clues as to the toxins present but more specific and informative assays are required. 7. Usually the additional AZA derivatives found in shellfish tissue are hydroxylated derivatives (e. and only a few structures (AZA 1 . minor revisions of a few stereocenters were made.9 Azaspiracids Azaspiracid (AZA1) was originally identified in 1995 as the causative agent for a poisoning episode in Ireland. Acetone extracts of fish tissue are subjected to solvent partitioning. and following synthesis of the molecule some time later.9). which are based on an LC-MS/MS approach. followed by clean-up on silica and aminopropyl cartridges.30%) and AZA3 (5 . Consequently. another in vitro sodium channel-based assay. Recently. almost 20 additional derivatives have been reported (83 . has been developed. and the separation of these polyethers from the lipid-soluble fraction of fish provides an additional challenge. and the corrected structure was published in 2004 (86) (Figure 7.80%). with AZA2 (10 . In this way Caribbean and Pacific CTX-1 compounds can be distinguished (74).9). specific for CTX activity. although specificity and sensitivity remain major hurdles (74) and the presence of DSP toxins affects the test (80).g.85). 84). and it is likely that most of these compounds are biotransformation products formed in the shellfish tissue. The original structure proposed revealed an entirely new class of shellfish toxin. Usually the major AZA derivatives found in shellfish are AZA1 (50 . Recent reviews of this new group of toxins have appeared (83. but this only has application in a clinical setting (81).
Following the original episode. Scandinavia. AZA toxins do 159 . While this result is not necessarily in debate. and to date this has been the only source from which these toxins have been found at toxic levels. cockles (Cerastoderma edule). scallops (Pecten maximus). Ostrea edulis). There has been considerable debate over the source of azaspiracids.9. However. and some time after the original episode in Ireland. Structure of the three principal azaspiracids found in shellfish 7. and off the coast of Morocco (83. and most recently in a crustacean (Cancer pagurus). clams (Tapes philipinarum.9. 90). Portugal.FISH AND SHELLFISH TOXINS Fig. these molluscs were never considered to be the biogenetic source. but current data indicates that unlike the DSP compounds. AZA occurrence has been reported by several European countries including France. 89). 7. 88. Ensis siliqua. which are ingested by P.2 Toxicology Consumption of AZA contaminated shellfish can result in acute symptoms that include nausea. cassipes (83). In fact the routes of trophic transfer of AZAs through the food chain ending in contaminated shellfish remain unknown. 7. Mytilus galloprovincialis).1 Origins and distribution The azaspiracids were first isolated from contaminated shellfish. it has been impossible to establish that P. The bivalves affected include mussels (Mytilus edulis. These symptoms are not unlike those experienced following the consumption of DSP toxins. 84. cassipes produces AZAs in culture. which can persist for 2 .3 days. Donax spp.9. as in other cases of shellfish toxins. diarrhoea and stomach cramps. it was reported that the heterotrophic dinoflagellate Protoperidinium cassipes was the planktonic source of the toxins (87).). and variable field results have led to suggestions that the actual producing organisms are Dinophysis spp. oysters (Crassostrea gigas. and to date no long-term effects have been recorded (83. vomiting.
This common cyclic imine feature is believed to be a common pharmacophore of this group of toxins which comprises the gymnodimines. and ASP. Using the same approach. but unlike DSP monitoring methods which use only the hepatopancreas of the shellfish for extraction. PSP. the spirolides.10 Cyclic imines These toxins are represented by several distinct structural types.3 Detection methods The regulatory level for AZAs in shellfish has been set by the EU at 160 µg AZA equivalents/kg shellfish tissue. 7. interferences can arise when analysing a matrix as complex as a crude shellfish extract and it is recommended that a simple solid phase extraction (SPE) step be introduced prior to LC/MS analysis (93). 92). but all contain a spirocyclic or cyclic imine function within their structure.FISH AND SEAFOOD not inhibit the phosphatases PP1 and PP2. However. 85). 91. The mouse bioassay has been used to monitor shellfish extracts.10 – 7. it is believed that the hydroxylated derivatives AZA4 (470 mg/kg) and AZA5 (<1000 mg/kg) are considerably less toxic.12). 7. AZAs also display cytotoxicity towards various mammalian cell lines (83. although autopsies conducted after 4 and 8 hours revealed various gastrointestinal (GI) abnormalities including accumulation of fluid in the ileum. though the possibility exists that they may inhibit other phosphatases. and erosion and bleeding in the stomach. it is suggested that the order of toxicity is AZA2 (110 µg/kg) >AZA3 (140 µg/kg) >AZA (150 µg/kg). Key fragment ions are useful in identifying the various AZA compounds that can occur in shellfish tissue (83. and closely related ptierotoxins (Figures 7. parenchymal cell pyknosis in the pancreas. Injection of a lethal dose of AZA1 in mice (>150 µg/kg) caused vacuole formation and fatty acid accumulation in hepatocytes. The limited amounts of AZA toxins available continue to hinder a complete understanding of their mechanism of action. Due to their relatively recent discovery and limited knowledge of their toxicological properties and impact on human 160 . Based on IP minimum lethal dose data.9. Oral administration of AZAs at substantially higher doses than those used in IP injection studies did not result in any mouse mortalities after 24 hours. abnormalities in the thymus and spleen. 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. which are particularly amenable to detection by positive ion electrospray mass spectrometry. Importantly these pathological outcomes were reported as being different from those caused by DSP. and necrosis of epithelial cells on the microvilli (91). the pinnatoxins. Currently LC/MS and LC-MS/MS analysis is the preferred chemical analytical detection method for AZAs. Once again. 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.
. 161 . Structure of the cyclic imine gymnodimine 7. 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. but they are included here for completeness.FISH AND SHELLFISH TOXINS health. 7. selliformis (97). surf clam and New Zealand abalone. led to the identification of a new marine toxin with an unprecedented structure named gymnodimine (96) (Figure 7.520 Da (94. scallops. gymnodimine B and C were isolated from the dinoflagellate Karenia selliformis (formerly Gymnodinium selliformis).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. it was proposed that a dinoflagellate identified as Gymnodinium spp. two additional congeners. In only two cases (gymnodimines and spirolides) has a planktonic source been identified.10). cockles. Fig. Some time later. 95).10. Pacific oyster. Subsequent taxonomic studies revised this again to a new dinoflagellate K. they are sometimes referred to as emerging toxins of uncertain human health concern. a species closely related to Gymnodinium mikimoitoi was the planktonic source of the toxin. 95). 7. At the time of the initial observation of gymnodimine.10. Reviews of the current status of the cyclic imine group were published by Molgo and Cembella (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.10.1. The structure is novel in that it contains a spirocylic imine group and a single tetrahydrofuran ring embedded within a macrocyclic structure.
95). A later study using purified toxin established an LD50 dose (Lethal Dose. Reduction of the imine group eradicated the toxicity. extracts exhibited the fast acting symptoms that became associated with cyclic imines following IP injection.11). The original observation of toxicity in New Zealand followed from IP injection of lipophilic extracts in the mouse bioassay. finishing with 100% methanol. together with a lack of evidence of human intoxication. Fractions are then dried and re-suspended in methanol or acetonitrile in preparation for LC/MS analysis (94. The structure of the spirolides bears a strong resemblance to both gymnodimine and 162 . cyclic imines are readily observed by electrospray mass spectrometry in the positive ion mode. almost a dozen derivatives in total have been identified to date (94.1. 95) (Figure 7. and so spectroscopic detection by UV methods is not favoured. Most cyclic imines lack an extended chromophore. edulis) and scallops (Placopectin magellanicus) from eastern Canada that displayed abnormal symptoms in the mouse bioassay. In particular. 50%) of 96 µg/kg.100). Oral administration of the toxin to mice by gavage resulted in about an 8.900 µg/kg). Based on this diminished oral toxicity. and this is the detection method of choice. However. and there was a further ten-fold reduction in toxicity when the toxin was supplied with food. This led to the suggestion that gymnodimine acts by blocking nicotinic acetylcholine receptors at the neuromuscular junction.10. 7. The time lapse between ingestion and death is between 5 and 15 minutes. and lack of responsiveness to stimuli. Since their original discovery which resulted in the characterisation of spirolides A-F (99. They were originally discovered in extracts of mussels (M.10.2 Toxicology A review of the toxicology of all cyclic imines has been published (98).1. abdominal breathing. and short-acting cholinesterase inhibitors such as neostigmine or physostigmine protected mice against the toxic effects of gymnodimine. 7. the first step in the process is typically an initial clean-up step using a short C18 cartridge and elution with water containing increasing amounts of methanol.FISH AND SEAFOOD 7.3 Detection methods The characteristic “fast-acting.10. all or nothing” mouse bioassay response to gymnodimines and cyclic imines in general continues to be a useful monitoring approach. paralysis of the hind quarters and eventually complete immobility. Whether isolated from dinoflagellate cells directly or from shellfish tissue.to10-fold reduction of toxicity (LD50 600 . There is a rapid onset of symptoms including respiratory distress.2 Spirolides The spirolides comprise the largest group of cyclic imine compounds. it has been stated that regulation against gymnodimine is not required (98).
7. southern Ireland. The LD50 values vary considerably depending upon whether the compound is administered by IP injection (5 .707 µg/kg. in fjords in Denmark and Norway. these are the toxins that are also most resistant to acid hydrolysis of the imine function 163 .10. fasted mice). the western coast of Scotland.FISH AND SHELLFISH TOXINS the pinnatoxins (vide infra). 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.11. by gavage (87 . Fig.2. Interestingly. fasted mice). or by oral ingestion (381 . This proved to be the case and eventually the dinoflagellate Alexandrium ostenfeldii was identified as the culprit organism (101). the gulf of Maine. All data reported here are for desmethyl spirolide C. 95).8 µg/kg). They have also been reported in the southern hemisphere in Chilean mussels and in New Zealand shellfish and plankton (94. their localisation in the digestive gland of mussels and scallops led to the suspicion that plankton were the real source of the toxins. While oral toxicity is considerably less than by IP injection. Since then the spirolides have been found to be globally distributed and are found in shellfish or plankton fractions along eastern Canada.1 Origins and distribution At the time of the original discovery of the spirolides in shellfish from the coastal waters of eastern Nova Scotia.166 µg/kg.2. Structure of four spirolide toxins A-D 7.10. those spirolides possessing the vicinal dimethyl grouping (see Figure 7.2 Toxicology The spirolides are the most studied group of the cyclic imines. the Bay of Biscay and most recently from Mediterranean shellfish and plankton in the Adriatic (102). R1 = CH3) of the cyclic imine ring are the most toxic of the group (98). 7.11.
which includes two diastereomers. 102). the mouse bioassay can be used to detect these fast acting toxins. 95. more useful information on the amount and distribution of toxins present in a shellfish extract is obtained by LC-MS analysis usually following a simple clean-up step. Fig.10.10. However. Structure of the pinnatoxin group and the closely related pteriatoxins 164 . Three members of this group have been reported.12. The compounds are zwitterionic and therefore somewhat more polar than the spirolides. While the mechanism of action remains to be clearly defined.12). 7.3 Pinnatoxins and pteriatoxins Pinnatoxins A .FISH AND SEAFOOD (100). Once again this relies on the sensitivity of these cyclic imines to detection by positive ion electrospray mass spectrometry. The pteriatoxins were isolated from a different mollusc but bear a strong structural resemblance to the pinnatoxins (Figure 7.B contain a spiro imine moiety similar to that found in the spirolides. and in fact these compounds share a number of other structural features.2. A number of methods following this procedure have been described (94. four members of the group have been identified – two of them being the C-34 diastereomers.3 Detection methods As discussed for gymnodimine. 7. To date. 7. consistent with modulation of acetyl cholinergic receptors (103). experiments have shown upregulation of muscarinic and nicotinic receptors in mice following dosing with spirolide C.
while pinnatoxin D was essentially inactive. 7.180 µg/kg).10.10. to date action or 165 . The LD99 dose for the pteriatoxins is reported to be in the range (8 .3 Detection methods No formal detection methods for these compounds have been reported.FISH AND SHELLFISH TOXINS 7. which occupy tropical waters around Okinawa and the south east coast of China.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.2 Toxicology There is little information describing the toxicology of these compounds (98). has come the realisation that a “toxin profile” can be a complex mixture of many different classes of toxins. 7.most likely a dinoflagellate.10. have long been suspected of causing human illness after ingestion. Coupled with this. 7.11 Conclusion There has been considerable progress in the detection of known marine toxins. 99%) values for pinnatoxins A-C are in the range (22 . The LD99 (Lethal Dose. With increasing knowledge of the chemistry of toxins. This is generally a result of increased monitoring and continually improving detection technologies and methods. This prompted a chemical investigation of Pinna muricata that ultimately led to the identification of the pinnatoxins in 1995 (104). and more advanced means of detection. new toxin groups such as the azaspiracids and cyclic imines have also been reported. which in many cases are now found to be more widespread than previously believed.3. While this is recognised as a real possibility for a wide range of toxins.3. and in some cases it is known that one toxin can potentiate or augment the toxicity of the other. there is no compelling evidence to suggest that they have resulted in human toxicity (98).3. There is strong circumstantial evidence to suspect that these toxins. but this remains to be conclusively established (106). like the gymnodimines and spirolides. arise from an algal source . Only the acute toxicity in mice following IP injection has been reported. and later the closely related pteriatoxins were isolated from the digestive glands of another Okinawan bivalve Pteria penguin (105). once again reflecting improvements in purification and spectroscopic methods permitting structural identification of new and complex structures.100 µg/kg). However. research and certified standards of the key toxins are often unavailable or in short supply and in the future considerable logistical effort will be required to address this issue. Despite this progress.
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.G.. Boca Raton. 131.. Abraham W. Food and Agriculture Organisation.. in Seafood and Freshwater Toxins. Ed.. Dickey R. Botana L..C. Brevetoxins. 70. Neurologic Shellfish Poisoning (NSP). Lenzi A. Poli M.. Marine Biotoxins.M. Flewelling L. 825-34. Boca Raton. Competitive ELISA to detect brevetoxins from Karenia brevis (formerly Gymnodinium breve) in seawater.. 2007. Bienfang P.. Johnson D. 2000.. Botana L. Abbott J. Fleming L.. Ramsdell J.D..R. shellfish and mammalian body fluid.B. Prockop L. Wang Z. CRC Press. Botana L. 67. CRC Press. Ciguatera fish poisoning: treatment prevention and management. 2008. Squicciarini D.. 74. and detection. 465-70. Benson J. 110.M.E. 2002. Watkins S. Kubanek J. Kubanek J. Pierce R..J. 71. Kirkpatrick B. Dickey R. Granade R. Locatelli C.. Harmful Algae. Fleming L..G. 171 . 2004.. 179-85..S. 6. Lancaster J...A. Toxicon. 2004. Naar J. Detection of brevetoxins in the twenty-first century.. Toxicon.H. West Indian Medical Journal. Baden D. Bourdelais A.. Boca Raton. 23. Bean J...S. like ciguatoxins. Buonocore M. 53. CRC Press.A. Fleming L.P.P. Food and Agriculture Organisation of the United Nations. Poon-King T. Ayyar R. Ciguatera toxins: Chemistry.. Steidinger K. 117-71. Ed. 187-94. Bourdelais A. 2004. Mild ciguatera poisoning: case reports with neurophysiological evaluations.M. 69.. toxicology. Backer L.... Zaias J. Butera R. Ciguatera fish poisoning in industrial ship crew members: a retrospective study in a seaport general practice in Trinidad and Tobago. New fish-killing alga in coastal Delaware produces neurotoxins. Environmental Health Perspectives.G. Cheng Y. 2008. 519-50..M. 2008.E. Food and Agriculture Organisation. Freidman M.. Baden D. Reich A. Dechraoui M-Y.. 68.. 50.. 75.E. 66. Ramsdell J. Ed... Flewelling L. Whitney P.. 72. Environmental Health Perspectives... Weisman R. Gannon D. Muscle Nerve.S. Ed. 73. Naar J. Marine Drugs. Clark R. Abraham W. Wanner A. in FAO Food and Nutrition Paper 80.. 1598-603. Manzo L.S.G.. Pierce R..E. 2008... Baden D... Landsberg J... Zaias J.A. Tomas C. 456-79. Schrank K. Baden D. Bottein M-Y.E... Fernandez M. Granholm A. Baden D...A.C. Naar J...G..N.J. 3. Chen A..M. 551-8. 479-516. 2007. Bossart G. in Seafood and Freshwater Toxins.. 99-115. The molecular and integrative basis to brevetoxin toxicity. Rome. Kirkpatrick B. Chest. Wolny J. 77. Pierce R. 2002.... Tomas C. 50.FISH AND SHELLFISH TOXINS 65.P. Reich A. in Seafood and Freshwater Toxins.. Gandini C.. Henry M. Intrinsic potency for synthetically prepared brevetoxin cysteine metabolites BTX-B2 and desoxy-BTX-B2. 76. Naar J. Jacocks H. Backer L. Aerosolised red tide toxins (Brevetoxins) and asthma.. 2007. Cheng Y. 110. 220-6..D.. Backer L.. 707-23. are potent ichthyotoxic neurotoxins that accumulate in fish. Poon-King C.W. Literature review of Florida red tide: implications for human health..
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.
Establish documentation concerning all procedures and records appropriate to these principles and their application. ingredient and packaging buyers. Establish a system to monitor control of the CCP. Establish the critical limit(s). The team should include members familiar with all aspects of the production process as well as specialists with expertise in particular areas such as production. 3.3 Stages of a HACCP Study The HACCP system consists of the following seven basic principles: 1. 4. Figure 8. quality assurance or control. An effective HACCP plan is best carried out as a multidisciplinary team exercise to ensure that the appropriate product-specific expertise is available. 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.A point. in addition to monitoring to determine compliance with the HACCP plan.Obtaining evidence that the elements of the HACCP plan are effective. tests and other evaluations. Determine the CCPs. Verification .1 is a flow diagram illustrating this 12-stage logic sequence. procedure. 7. Conduct a hazard analysis. Each stage is discussed below in detail. food microbiology. 5. Establish the corrective action to be taken when monitoring shows that a CCP is not under control. 8.The application of methods. operation or stage in the food chain including raw materials.HACCP IN FISH AND SEAFOOD PRODUCT MANUFACTURE Step .3. hygiene managers.1 Assemble HACCP team HACCP requires management commitment of resources to the process. procedures. food chemistry or engineering. from primary production to final consumption. Establish procedures for verification to confirm that the HACCP system is working effectively. The team should also include 177 . 6. Validation . 2. 8.
chemical and/or physical). The scope of the plan should be determined by defining the extent of the production process to be considered and the categories of hazard to be addressed (e.g. it may be obtained from external sources. Logic sequence for application of HACCP 178 . If expert advice is not available on-site.1. Fig. biological.FISH AND SEAFOOD personnel who are involved with the variability and limitations of the operations. 8.
The description should include information such as the product name. shelf life.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. naturally present on fish from warm waters (> 15 °C).).3. contaminated fish and shellfish should not be used. storage and distribution conditions and instructions for use. marine toxins associated with particular fish and shellfish species. If the processing does not include a kill step.3. physical and chemical structure (including water activity (aw). aquaculture drug residues and potentially pathogenic Vibrio spp. fermentation. there may be species-specific hazards related to certain fish and shellfish. These factors are likely to have a significant effect on some potential hazards.1 Fish and seafood The HACCP team should ideally have access to expertise on the appropriate fisheries.1. the only CCP that can render the product safe from pathogenic organisms is adequate heat treatment during preparation. It should provide an accurate representation of all steps in the production process from raw materials to the end179 .2 Describe the product It is important to have a complete understanding of the product.3. including practices onboard fishing vessels. or in aquaculture operations. such as children or the elderly.g. packaging.g.). However. such as enteric viruses in shellfish. is a cooking process required?). etc. Vulnerable groups of the population. 8. may need to be considered specifically. 8. processing conditions (e. etc. composition.3. In addition. for example contamination by foodborne pathogens as a result of time/temperature abuse or unsatisfactory handling practice. 8.1 Fish and seafood Certain products may be contaminated or carry pathogenic organisms as a part of the natural flora. It is also important to identify the consumer target groups.3.3 Identify intended use The intended use should be based on the expected uses of the product by the enduser or consumer (e. it must be noted here that certain toxins (algal and shellfish) are not destroyed by heat treatments. 8. freezing.HACCP IN FISH AND SEAFOOD PRODUCT MANUFACTURE 8.3. which should be described in detail. pH. Expertise on such inherent hazards. challenge testing and inoculation studies for evaluation of safety aspects are therefore essential for an effective HACCP team. heat treatment.
The operation should be observed at all stages.FISH AND SEAFOOD product. or circumstances of any reclaim or rework activity. Many fish-processing operations have relatively few steps and the flow diagrams appear simple.6 List all potential hazards associated with each step. time/temperature data.3. cleaning and hygiene procedures and storage conditions. conduct a hazard analysis. survival or multiplication of pathogenic microorganisms. The analysis is likely to include consideration of: ● the likely occurrence of hazards and the severity of their adverse health effects. once determined. Ideally. and identify any measures to control identified hazards The HACCP team should list all hazards that may reasonably be expected to occur at each step in the production process. and therefore a detailed description of the procedure applied must be available.1 Fish and seafood Examples of flow diagrams for specific fish products may be found in the appropriate product chapters. it should also include details of CCP steps. 180 ● ● .4. It is also important to include observation of production outside normal working hours such as shift patterns and weekend working. adequate chilling of finfish immediately after capture is vital for the control of several potential hazards. 8. it is essential that the details of each step are fully appreciated and recorded. It may include details of the factory and equipment layout. features of equipment design. It is essential that the diagram is accurate. For example. 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. the qualitative and/or quantitative evaluation of the presence of hazards. then any pre-drawn HACCP plans must be reviewed once the lines/products are finalised. ingredient specifications. 8.3.3. and any discrepancies between the flow diagram and normal practice must be recorded and the diagram amended accordingly. because the hazard analysis and decisions regarding CCPs are based on these data. However. If HACCP studies are applied to proposed new process lines or products.
or poor control of batter mixes in frozen battered fish portions may allow growth of Staphylococcus aureus and production of enterotoxin. Note: it is important at this stage that no attempt is made to identify CCPs. One control measure may be relevant to several process steps. Many of the microbiological hazards associated with fish and seafood products are derived from the raw materials. The incidence of psychrotrophic.).3. modified-atmosphere packaging (MAP) of chilled raw fish may provide conditions suitable for the growth of psychrotrophic C. Pathogens may be part of the resident microflora of the living animal (e. since this may interfere with the analysis. and viruses). For example. Hazards introduced during processing of fish products depend very much on the characteristics of the process. Salmonella spp.1 Fish and seafood The term 'fish and seafood' includes an extremely varied group of products. or may originate from polluted water or from post-capture contamination (e. there are particular hazards associated with contamination of shellfish by microorganisms from human sewage. The HACCP team should then determine what control measures exist that can be applied for each hazard.g. Vibrio spp. or desirable. Some hazards may require more than one control measure for adequate control and a single control measure may act to control more than one hazard.g. Therefore it is not possible.6. and there are an equally varied range of potential hazards associated with them. the hurdle effect. any food safety objectives or manufacturer’s food safety requirements.HACCP IN FISH AND SEAFOOD PRODUCT MANUFACTURE ● ● ● ● production or persistence of toxins. botulinum. 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. where a hazard is repeated. the number of consumers potentially exposed and their vulnerability. and the potential growth of Listeria monocytogenes on smoked fish. Hazards specific to certain types of product are detailed in the appropriate chapters of this manual. 8. to generalise about expected hazards and the reader is once again referred to the appropriate product chapter for additional advice on specific hazards. For example. non-proteolytic Clostridium botulinum on fresh fish is also sufficiently high that its presence may be assumed. 181 .
critical limits should be specified and validated for each CCP. structured approach to decision making.FISH AND SEAFOOD 8. and its use may not always be appropriate. it can be said that effective control measures are likely to include the following: • • • • • 8.7. Where possible. 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. 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 . then the process must be modified to include a control measure.7 Determine CCPs The determination of CCPs in the HACCP system is facilitated by a decision tree (Figure 8. It is prudent to set stricter limits (often-called target or process limits/levels) to ensure that any trend towards a loss of control is noted before the critical limit is exceeded.3. but for which no control exists at that step or any other. More than one critical limit may be defined for a single step.1 Fish and seafood Specific product chapters provide information on criteria that may be used to set critical limits. application of the decision tree should be flexible. 8. Criteria used to set critical limits must be measurable and may include physical.3. However. biological or sensory parameters. it is necessary to specify both time and temperature for a thermal process. However. It is also essential that the HACCP team has access to sufficient technical data to determine the CCPs effectively.2) to provide a logical.8.3.3. For example. If a significant hazard has been identified at a step where control is required for safety. and the reader is referred to the appropriate product chapter. chemical. given the enormous variety of fish products and processes in use.1 Fish and seafood Again. 8.
HACCP IN FISH AND SEAFOOD PRODUCT MANUFACTURE
• • • 8.3.9
Water activity (aw) values for dried fish pH values in fermented fish Microbiological quality of water in shellfish production areas Establish a monitoring system for each CCP
Monitoring involves planned measurement or observation of a CCP relative to its critical limits. Monitoring procedures for target levels must be able to detect loss of control of the CCP, and should provide this information with sufficient speed to allow adjustments to be made to the control of the process before the critical limits are violated. Monitoring at critical limits should be able to rapidly detect when the critical limit has been exceeded. Monitoring should therefore either be continuous, or carried out sufficiently frequently to ensure control at the CCP. Therefore, physical and chemical on-line measurements are usually preferred to lengthy microbiological testing. However, certain rapid methods, such as ATP assay by bioluminescence, may be useful for assessment of adequate cleaning, which could be a critical limit for some CCPs, for example pre-start-up hygiene. Persons engaged in monitoring activities must have sufficient knowledge, training and authority to act effectively on the basis of the data collected. These data should also be properly recorded. 8.3.10 Establish corrective actions
For each CCP in the HACCP plan, there must be specified corrective actions to be applied if the CCP is not under control. If monitoring indicates a deviation from the critical limit for a CCP, action must be taken that will bring it back under control. Actions taken should also include proper isolation of the affected product, repair of defective equipment, and an investigation into why the deviation occurred. A further set of corrective actions should relate to the target levels, if process drift is occurring. In this case, only repair of the process defect and investigation of the fault are required. All corrective actions should be properly recorded. 8.3.11 Establish verification procedures
Verification usually involves auditing and testing procedures. Auditing methods, procedures and tests should be used frequently enough to determine whether the HACCP system is being followed, and is effective at controlling the hazards. These may include random sampling and analysis, including microbiological testing. Although microbiological analysis is generally too slow for monitoring purposes, it can be of great value in verification, since many of the identified hazards are likely to be microbiological. In addition, reviews of HACCP records are important for verification purposes. These should confirm that CCPs are under control and should indicate the nature
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of any deviations and the actions that were taken in each case. It is also useful to review customer returns and complaints regularly. 8.3.12 Establish documentation and record keeping
Efficient and accurate record keeping is an essential element of a HACCP system. The procedures in the HACCP system should be documented. ● ● Determination of critical limits The completed HACCP plan
Examples of documented procedures include: ● ● The hazard analysis Determination of CCPs
Examples of recorded data include: ● ● ● Results of monitoring procedures Deviations from critical limits and corrective actions Records of certain verification activities, e.g. observations of monitoring activities, and calibration of equipment.
The degree of documentation required will depend partly on the size and complexity of the operation, but it is unlikely to be possible to demonstrate that an effective HACCP system is present without adequate documentation and records. The length of time that records are kept will be as per company policy, but should not be less than one year beyond the shelf life of the product. Three to five years is typical for many food companies. 8.4 Implementation and review of the HACCP plan
The completed plan can only be implemented successfully with the full support and co-operation of management and the workforce. Adequate training is essential and the responsibilities and tasks of the operating personnel at each CCP must be clearly defined.
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Fig. 8.2. CCP Decision Tree (Adapted from Codex Alimentarius Commission, 2003) Finally, it is essential that the HACCP plan be reviewed following any changes to the process, including changes to raw materials, processing conditions or equipment, packaging, cleaning procedures and any other factor that may have an effect on product safety. Even a small alteration to the product or process may invalidate the HACCP plan and introduce potential hazards. Therefore, the implications of any changes to the overall HACCP system must be fully
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considered and documented, and adjustments made to the procedures as necessary. Triggered reviews/audits should occur as a result of changes, whereas a scheduled review/audit should be annual, as a minimum. 8.5 References
Wareing P.W., Carnell A.C. HACCP – A Toolkit for Implementation. Leatherhead, Leatherhead Food International. 2007. Bernard D., Scott V. Hazard Analysis and Critical Control Point System: use in controlling microbiological hazards, in Food Microbiology: Fundamentals and Frontiers. Eds. Doyle M.P., Beuchat L.R. Washington D.C., ASM Press. 2007, 971-86. Su Y-C., Daeschel M.A. Sanitation and HACCP, in Surimi and Surimi Seafood. Ed. Park J.W. Boca Raton, CRC Press. 2005, 141-62. Huss H.H. Considerations in the application of the HACCP principles to seafood production, in Assessment and Management of Seafood Safety and Quality. Eds. Food and Agricultural Organisation, Huss H.H., Ababouch L., Gram L. Rome, FAO. 2004, 153-77. Chen H-C., Chang P.C-M. Frozen seafood safety and HACCP, in Handbook of Frozen Foods. Ed. Hui Y.H., Cornillon P., Legaretta I.G., Lim M.H., Murrell K.D., Nip WK. New York, Marcel Dekker. 2004, 339-63. Mortimore S., Mayes T. The effective implementation of HACCP systems in food processing, in Foodborne Pathogens: Hazards, Risk Analysis and Control. Eds. Blackburn C.deW., McClure P.J. Cambridge, Woodhead Publishing Ltd. 2002, 229-56. Mayes T., Mortimore C.A. Making the Most of HACCP: Learning from Other’s Experience. Cambridge, Woodhead Publishing. 2001. Mortimore S.E., Wallace C., Cassianos C. HACCP (Executive Briefing). London, Blackwell Science Ltd. 2001. Chartered Institute of Environmental Health. HACCP in Practice. London, Chadwick House Group Ltd. 2000. Jouve J.L. Good manufacturing practice, HACCP, and quality systems, in The Microbiological Safety and Quality of Food, Volume 2. Eds. Lund B.M., BairdParker T.C., Gould G.W. Gaithersburg, Aspen Publishers. 2000, 1627-55. Stevenson K.E., Bernard D.T. HACCP: A Systematic Approach to Food Safety. Washington D.C., Food Processors Institute. 1999. National Seafood HACCP Alliance. IQF cooked shrimp processing HACCP model, in HACCP User's Manual. Ed. Corlett D.A. Gaithersburg, Aspen Publishers. 1998, 454-73. Mortimore S., Wallace C. HACCP: A Practical Approach. Gaithersburg, Aspen Publishers. 1998.
HACCP IN FISH AND SEAFOOD PRODUCT MANUFACTURE Khandke S.S., Mayes T. HACCP implementation: a practical guide to the implementation of the HACCP plan. Food Control, 1998, 9 (2-3), 103-9. Forsythe S.J., Hayes P.R. Food Hygiene, Microbiology and HACCP. Gaithersburg. Aspen Publishers. 1998. Food and Agriculture Organisation. Food Quality and Safety Systems: A Training Manual on Food Hygiene and the Hazard Analysis and Critical Control Point (HACCP) System. Rome, FAO. 1998. US Food and Drug Administration. Fish and Fishery Products Hazards and Controls Guide. Rockville, Food Drug Administration. 1998. National Advisory Committee on Microbiological Criteria for Foods. Hazard Analysis and Critical Control Point Principles and Application Guidelines. 1997. Gardner L.A. Testing to fulfil HACCP (Hazard Analysis Critical Control Points) requirements: principles and examples. Journal of Dairy Science, 1997, 80 (12), 3453-7. Martin R.E., Collette R.L, Slavin J.W. Fish Inspection, Quality Control and HACCP: A Global Focus. Proceedings of a Conference, Arlington, May 1996. Lancaster P.A., Technomic Publishers. 1997. Codex Alimentarius Commission. Hazard Analysis Critical Control Point (HACCP) System and guidelines for its application, in Food Hygiene: Basic Texts. Ed. Codex Alimentarius Commission. Rome, FAO. 2003, 21-31. Garrett E.S., Jahncke M.l., Tennyson J.M. Microbiological hazards and emerging food safety issues associated with seafoods. Journal of Food Protection, 1997, 60 (11), 1409-15. National Seafood HACCP Alliance. HACCP: Hazard Analysis and Critical Control Point Training Curriculum. Raleigh, North Carolina State University. 1997. Dillon M., McEachern V. HACCP and quality assurance of seafood, in Fish Processing Technology. Ed. Hall G.M. London, Blackie. 1997, 224-48. Savage R.A. Hazard Analysis Critical Control Point: a review. Food Reviews International, 1995, 11 (4), 575-95. Pearson A.M., Dutson T.R. HACCP in Meat, Poultry and Fish Processing. Glasgow, Blackie. 1995. Huss H.H., Food and Agriculture Organisation. Assurance of Seafood Quality. FAO Fisheries Technical Paper No. 334. Rome, FAO. 1994. Bonnell A.D. Process control, in Quality Assurance in Seafood Processing: A Practical Guide. Ed. Bonnell A.D. London, Chapman & Hall. 1994, 39-53. Pierson M.D., Corlett D.A., Institute of Food Technologists. HACCP: Principles and Applications. New York, Van Nostrand Reinhold. 1992. Hudak-Roos M., Garrett E.S. Monitoring, corrective actions and record keeping in HACCP, in Quality Assurance in the Fish Industry: Proceedings of an International Conference, Copenhagen, August 1991. Eds. Huss H.H., Jakobsen M., Liston J. Amsterdam, Elsevier. 1992, 521-31.
1988. 33-44. Geneva. World Health Organisation. Microorganisms in Foods. International Commission on Microbiological Specifications for Foods. WHO.. Volume 4: Application of the Hazard Analysis Critical Control Point (HACCP) System to Ensure Microbiological Safety and Quality. 1992. International Journal of Food Microbiology. Mayes T. Oxford. 3 (1). 14-9. 1992. Development and use of the HACCP concept in fish processing.L. Hazard Analysis Critical Control Point Evaluations: A Guide to Identifying Hazards and Assessing Risks Associated with Food Preparation and Storage. 188 . Bryan F. Simple users' guide to the hazard analysis critical control point concept for the control of food microbiological safety. Blackwell Scientific Publications. Food Control. 15 (1/2).H. 1992.HACCP IN FISH AND SEAFOOD PRODUCT MANUFACTURE Huss H.
however. final product specifications. from structural conditions in the factory or process facility.9. Hygiene is a general concept that covers a wide subject area. and conditions of raw materials. The new legislation was designed to establish conditions. Microbiological standards have a useful role and help establish requirements for the microbiological safety and quality of food and raw materials. transport and delivery vehicles requirements. with the intention of introducing consistency and clarity throughout the food production chain from primary production. the existence of microbiological standards alone cannot protect consumer health. to sale or supply to the final consumer. eliminate or acceptably control pathogen contamination of food. and one that plays an important role in most countries’ food legislation. Jenny Pfleger Leatherhead Food International Randalls Road Leatherhead Surrey KT22 7RY United Kingdom 9. A number of standards are provided in food legislation. have been replaced by several linked measures on food safety rules and associated animal health controls. 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. 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. under which food is produced to optimise public health and to prevent. Procedures under the new legislation are based on risk assessment and management and follow a 'farm to fork' approach to food 189 . to personnel requirements.1 Introduction Hygiene is an important aspect of ensuring food safety. including microbiological criteria. EC FOOD HYGIENE LEGISLATION Eugenia Choi and Dr. A new package of European Commission (EC) hygiene measures became applicable on 1 January 2006 to update and consolidate the earlier 17 hygiene directives.
A number of more detailed implementing and transitional measures have been adopted at EC level. 24.1).9. 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. 7. p. European Communities L 268. p. Prescribed are detailed measures to ensure the safety and wholesomeness of food during preparation. handling and offering for sale or supply to the consumer. manufacturing. 9. Regulation 854/2004 relates to the organisation of official controls on products of animal origin. 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. 853/2004 of the European Parliament and of the Council laying down specific hygiene rules for food of animal origin Regulation (EC) No. 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. packaging.9.1992. 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. processing.7. 852/2004 of the European Parliament and of the Council on the hygiene of foodstuffs Regulation (EC) No. p. transportation.1991. the following European Union (EU) hygiene regulations have applied: • • • Regulation (EC) No. and sets out what those enforcing the provisions have to do. 41).1991. distribution. Subsequently. storing. 24. J. 190 • • . N.2 Legislative Structure From 1 January 2006.15). as last amended by Regulation (EC) No 806/2003. 2073/2005 on microbiological criteria for foodstuffs • The general hygiene requirements for all food business operators are laid down in Regulation 852/2004. as last amended by Regulation (EC) No 806/2003.FISH AND SEAFOOD safety with the inclusion of primary production in food hygiene legislation. Regulation 853/2004 supplements Regulation 852/2004 in that it lays down specific requirements for food businesses dealing with foods of animal origin.B.
processing and distribution of food under their control satisfy the relevant hygiene requirements laid down in Regulation (EC) No. Although EU food hygiene regulations are directly applicable in the individual Member States there are some aspects where Member States are required or allowed to adopt certain provisions into their national laws. 852/2004 on the General Hygiene of Foodstuffs Food business operators must ensure that all stages of production. particularly taking into account a number of factors ranging from ensuring food safety throughout the food chain to begin with primary production.I. food storage that is for private or domestic consumption. of stock farming. 852/2004. 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. Although the regulations have the force of law. As regulations.I. 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. 9. 191 .7. revocation of existing implementing legislation. Primary products are defined as products of primary production including products of the soil.14. penalties and powers of entry. and equivalent legislation in Scotland.) in England. and also if the producer supplies small amounts of primary product to the final consumer or local retail establishments supplying the final consumer. with primary production. 1 .3 Regulation (EC) No. and of hunting and fishing. 19. 2073/2005.1993. and binding in its entirety on all member states from the date of entry into force. Additionally the requirements of EC Regulation 853/2004 must be complied with. p. This Regulation lays down general rules for food business operators on the hygiene of foodstuffs. the legislation is directly applicable law.g.LEGISLATION • Council Directive 93/43/EEC of 14 June 1993 on the hygiene of foodstuffs (OJ L 175. The Food Hygiene (England) Regulations 2006 (S. which will be covered later in this chapter. Wales and Northern Ireland). 2006 No. Wales and Northern Ireland. right through to the implementation of procedures based on HACCP principles. setting offences. domestic preparation or handling. as amended) came into force on 11 January 2006 (separate but similar national legislation also came into force on that day in Scotland. national legislation in the form of a Statutory Instrument (S. etc. is required to give effect to the EU regulations. e.11) The EU hygiene regulations apply to all stages of food production including primary production. The national legislation in all four UK countries also applied the provisions of the EU Microbiological Criteria Regulation No. for example.
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Annex I – Primary production
Annex I (Part A) relates to general hygiene provisions for primary production and associated operations covering: (a) the transport, storage and handling of primary products at the place of production, (b) the transport of live animals, (c) for fishery products - transport operations to deliver primary products (which have not been substantially altered) from the place of production to an establishment Food business operators have the responsibility to ensure primary products are protected against contamination. Any community and national legislation relating to the control of hazards in primary production such as measures to control contamination resulting from surroundings e.g. air, soil, water etc., and measures relating to animal health and welfare, and plant health that may impact on human health should be complied with. Requirements for record keeping are also laid down. This relates to animal feed (nature and origin), veterinary medicines administered to animals (date given and withdrawal periods), any diseases, analysis of samples from other animals that might impact on human health as well as reports on animal checks performed. Part B of Annex I contains recommendations for guides to good hygiene practice. 9.3.2 Annex II – Stages other than primary production
Annex II of the regulation lays down additional general hygiene requirements that must be met by food business operators carrying out production, processing and distribution of food at all stages following primary production. A summary of Chapters I to IV of Annex II is provided as follows: 22.214.171.124 Chapter I Chapter I applies to all food premises, except premises to which Chapter III applies. Food premises must be kept clean and maintained in good repair and condition. Their layout should allow for this. The environment should allow good hygiene practices and provide temperature-controlled handling and storage conditions where necessary ,and enable foods to be kept at correct temperatures and be monitored.
Additionally, there are requirements for provision of adequate lavatories, basins, ventilation, lighting and drainage.
126.96.36.199 Chapter II Chapter II applies to all rooms where food is prepared, treated or processed, except dining areas and premises to which Chapter III applies. The design and layout of rooms should allow for good hygiene practices between and during operations. Therefore floor and wall surfaces, ceilings and windows should be constructed to prevent dirt accumulating. Surfaces where food is handled must be well-maintained and allow easy cleaning and disinfection, preferably using smooth, washable corrosionresistant and non-toxic materials. There should be facilities for cleaning or disinfecting, and for storing working utensils or equipment. Clean potable water and adequate provision for washing food is needed.
188.8.131.52 Chapter III Chapter III applies to temporary premises (e.g. marquees, market stalls, and mobile sales vehicles), premises used primarily as a private dwelling-house but where foods are regularly prepared for placing on the market, and vending machines. Premises and vending machines should be practically sited, designed, constructed and kept clean and maintained in good repair and condition so as to avoid the risk of contamination, in particular by animals and pests. Facilities should allow adequate personal hygiene, and surfaces in contact with food should be easy to clean. Enough potable water and storage arrangements for hazardous or inedible substances are required as well as adherence to food safety requirements.
184.108.40.206 Chapter IV Chapter IV applies to all transportation. Conveyances and/or containers used for transporting foodstuffs are to be kept clean and maintained in good repair and condition to protect foodstuffs from contamination and are, where necessary, to be designed and constructed to permit adequate cleaning and/or disinfection. Food should be maintained at appropriate temperatures.
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220.127.116.11 Chapter V Chapter V refers to equipment requirements. Adequate cleaning and disinfection is to be carried out frequently for articles, fittings and equipment in contact with food where contamination needs to be avoided. Equipment should be installed to allow adequate cleaning, and be fitted with the required control device.
18.104.22.168 Chapter VI Chapter VI refers to food waste. Food waste, non-edible by-products and other refuse is to be removed from rooms where food is present as quickly as possible to avoid accumulation. Such waste is to be deposited in closable containers, to allow easy cleaning. Refuse stores should allow easy cleaning and be free of pests. Waste must be eliminated hygienically in accordance with European Community legislation.
22.214.171.124 Chapter VII Chapter VII refers to the water supply. There must be an adequate supply of potable water, which should be used whenever necessary to ensure foodstuffs are not contaminated. There are also requirements for recycled water, ice in contact with food, use of steam and for the water used in the cooling process for heat-treated foods in hermetically sealed containers. Specific requirements are that clean water may be used with whole fishery products. Clean seawater, may be used for live bivalve molluscs, echinoderms, tunicates and marine gastropods; and for external washing. If it is used, there must be adequate facilities to ensure the supply does not create a source of contamination for the foodstuff. Recycled water used in processing or as an ingredient must not present a contamination risk. It must be of the same standard as potable water, unless the competent authority is satisfied that the quality of water cannot affect the wholesomeness of food in the finished form.
Additionally, ice that comes into contact with food or which could possibly contaminate it needs to be made from potable water or, in the case of chilling whole fishery products using clean water. Ice must be made, handled and stored under conditions that protect it from contamination.
126.96.36.199 Chapter VIII Chapter VIII sets out personal hygiene requirements for those working in a food handling area including clean protective clothing and the requirement that those carrying or suffering from a disease are not permitted to handle food. 188.8.131.52 Chapter IX Chapter IX covers provisions applicable to foodstuffs. A food business operator should not accept raw materials or ingredients, other than live animals, or any other material used in processing products, if they are known to be contaminated with parasites, pathogenic microorganisms or foreign substances, to be toxic or decomposed to such an extent that, even after the business operator applied normal hygienic processing , the product would be inedible. Raw materials must be kept under appropriate conditions throughout production, processing and distribution. In particular, temperature control (i.e. cold chain and food thawing) requirements are laid down.
184.108.40.206 Chapter X Chapter X lays down provisions applicable to the wrapping and packaging of foodstuffs to avoid contamination of any form. 220.127.116.11 Chapter XI Chapter XI lays down heat treatment requirements for food that is placed on the market in hermetically sealed containers. The main relevant process parameters (particularly temperature, pressure, sealing and microbiology) must be checked by the food business operator to ensure that the required heat treatment has been achieved. Also, the process used should comply with an internationally recognised standard (e.g. pasteurisation, ultra high temperature or sterilisation). 18.104.22.168 Chapter XII Chapter XII states training requirements for food business operators to ensure that food handlers are trained in food hygiene matters and in the application of HACCP principles.
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The Regulation requires that food business operators must notify the relevant competent authority of each establishment under their control that carries out any of the stages of production, processing and distribution of food, with a view to the registration of each establishment. Food business operators must also ensure that the competent authority always has up-to-date information on establishments, including the notification of significant changes in activity, and closure of an existing establishment. Food business operators must ensure that establishments are approved by the competent authority, following at least one on-site visit, when approval is required by the national law of the Member State in which the establishment is located, or under Regulation (EC) No. 853/2004, or by a separate decision adopted. Separate rules apply for businesses producing products of animal origin. 9.3.4 HACCP
Food business operators, other than at the level of primary production, and associated operations must put in place, implement and maintain a permanent procedure or procedures based on principles of the system of hazard analysis and critical control points (HACCP). Emphasis is placed on risk-related control, with responsibility placed on the proprietor of the food business to ensure that potential hazards are identified and systems are developed to control them. Under HACCP, food business operators must amongst other considerations, identify hazards to be prevented or eliminated or reduced to acceptable levels, identify critical control points (CCP) at which control is essential to prevent, eliminate or reduce hazards and establish critical limits at these points, implement effective monitoring procedures, and establish corrective actions in the case where a CCP is out of control. Procedures must be followed to confirm the above is in place and up-todate, as well as providing the competent authority with documents and records as evidence, when required. 9.4 Regulation (EC) No. 853/2004 Laying Down Specific Hygiene Rules for Food of Animal Origin
Regulation (EC) No. 853/2004 lays down hygiene rules for products of animal origin that apply in addition to the general hygiene rules of Regulation (EC) No. 852/2004. 9.4.1 Definitions
The following definitions apply for fishery products and live bivalve molluscs:
skinning. slime or sand. 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. shelling. slicing. purification centres or dispatch centres in tanks or any other installation containing clean seawater. and all mammals. reptiles and frogs) whether wild or farmed and including all edible forms. including products packaged under vacuum or in a modifiedatmosphere that have not undergone any treatment to ensure preservation other than chilling. Prepared fishery products means unprocessed fishery products that have undergone an operation affecting their anatomical wholeness. followed by wrapping or packaging. Bivalve molluscs means filter-feeding lamellibranch molluscs Marine biotoxins means poisonous substances accumulated by bivalve molluscs. Freezer vessel means any vessel on board in which freezing of fishery products is carried out. heading. to preserve or to improve organoleptic qualities and to ensure they are in a good state of vitality before wrapping or packing.LEGISLATION Fishery products means all seawater or freshwater animals (except for live bivalve molluscs. where necessary. or in natural sites. Mechanically separated fishery product means any product obtained by removing flesh from fishery products using mechanical means resulting in the loss or modification of the flesh structure. in particular as a result of feeding on plankton containing toxins. and chopping. after preparatory work such as bleeding. mincing or processing. to remove mud. slicing. live echinoderms. shucking. heading. if necessary: chilling or freezing: filleting. Fresh fishery products means unprocessed fishery products. parts and products of such animals. live tunicates and live marine gastropods. 197 . gutting and removal of fins and. filleting. Factory vessel means any vessel on board which fishery products undergo one or more of the following operations followed by wrapping or packaging and. such as gutting. Conditioning means the storage of live bivalve molluscs coming from class A production areas. whether whole or prepared. where appropriate.
(ii) However.4. EG. EB or WE. (iii) Food business operators must have in place systems and procedures to identify food business operators from whom they have received and to whom they have delivered products of animal origin. EO. EK. If an establishment manufactures both food to which Regulation 853/2004 applies and food to which it does not. ES.1 Identification marking An identification mark needs to be applied following the requirements of Annex II. EY. 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. EK. the mark must show the approval number of the establishment. the code would be UK. Extracts of the requirements of Annex II. In this case.1.2. a new mark must be applied to the product.1. and the characters easily decipherable and be clearly displayed. the food business operator may apply the same identification mark to both types of food.2 Fishery product requirements 9.2 Form of the identification mark (i) The mark must be legible and indelible.4. please refer to Regulation (EC) 853/2004.2. 9. (iii) Additionally. the mark must be oval shaped and include the abbreviation CE. the new mark must show the approval number of the establishment where this occurred. for other codes.4. 198 . (iv) If the identification mark is applied in an establishment within the Community. (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. EF.FISH AND SEAFOOD 9. EÜ. Section I of Regulation (EC) 853/2004 are given as follows: 9. For the UK. Section I of Regulation (EC) 853/2004. when a product's packaging and/or wrapping is removed or is further processed in another establishment.4.2. EC.1 Application of the identification mark (i) The identification mark must be applied before the product leaves the establishment of production.
Section VIII of Regulation 853/2004. they would need to be obtained following Section VII on live bivalve molluscs. This section does not apply to bivalve molluscs.2. and are intended for further handling. the requirements accompany those of Annex II of Regulation (EC) 852/2004 199 . reference should be made to the actual Regulation. and fishery products carried in bulk. With the exception of Chapters I and II. 9. the approval number of the establishment and the abbreviation for the community where appropriate. the wrapping or the packaging. (v) When the mark is applied directly to products of animal origin.1.LEGISLATION 9. as amended. including vessels. specifically relating to fishery products are given below. the mark may be applied to the outer surface of the container or packaging. tunicates and marine gastropods when placed on the market live. (iii) In the case of liquid. echinoderms. or be printed on a label affixed to the product. Chapter III. processing. It may also be a tag made of a resistant material affixed in such a way that it cannot be removed. it is sufficient to apply the mark to the exterior of that package only. The requirements of this section accompany those in Regulation (EC) 852/2004. the colourings used must be EC authorised colours. wrapping or packaging in another establishment.2. it applies to those animals when not placed on the market live. Parts A. Chapter IV. an identification mark is not necessary if accompanying documentation contains the name of the country or its two-letter code. Part A and Chapter V apply to retail. a) requirements for establishments. i. C and D. its wrapping or the packaging.4.4. (ii) For products of animal origin that are placed in transport containers or large packages. involved in primary production and such operations accompany the needs of Annex I of Regulation (EC) 852/2004 b) for other establishments including vessels.e. for full requirements. Therefore. (iv) If products of animal origin are placed in a package destined for direct supply to the final consumer. Extracts of the requirements of Annex III.3 Method of marking (i) The mark is to be applied directly to the product. granulate and powdered products of animal origin carried in bulk.2 Hygiene requirements for fishery products The regulation details specific hygiene requirements for fishery products.
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. There is a derogation from point a. gutting. 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.e. from the place of production to the first establishment of destination. it must be placed somewhere that will not contaminate the water supply. they also include (i) the transport and storage of fishery products that have not been substantially altered (including live fishery products.) 9.4. Chapter VII of Regulation 852/2004. The definition of fishery products covers primary production as well as its associated operations. i. removing fins. surfaces that contact fishery products need to be made from cleanable corrosion-resistant material.2. heading. 2.. Requirements for all vessels Vessels should be made in a way that does not cause contamination with substances such as fuel and oil etc. or to handle or process them after harvesting. Requirements for vessels designed and equipped to preserve fresh fishery products for more than 24 hours 200 .2. bleeding. fishing and collection of live fishery products with a view to their being placed on the market. and (b) related operations include any of the following operations if done on board fishing vessels: slaughter.FISH AND SEAFOOD c) for the water supply.1 Chapter I . if a vessel has water coming in that is used with fishery products. Structural and equipment requirements refer to the following points 1. A. the production of ice used to chill fishery products and the rapid cooling of crustaceans and molluscs after their cooking. (a) primary production covers the farming. this supplements the requirements of Annex II. clean seawater may be used for the handling and washing of fishery products. within fish farms on land) (ii) the transport of fishery products that have not been substantially altered (including live fishery products.
201 . and not more than 0 °C after 16 hours. Storage areas for the finished products also need to be sufficiently large and cleanable. 3. (ii) Refrigeration equipment needs to keep fishery products in storage holds at not more than -18 °C.LEGISLATION (i) Such vessels need to have holds. Also. Holds and containers used for storing fishery products must allow its preservation under adequate hygiene conditions and. 4. (iii) In vessels equipped for chilling fishery products in cooled clean seawater. where necessary. Such devices must achieve a chilling rate that ensures that the mix of fish and clean seawater does not exceed 3 °C six hours after loading. tanks or containers for the storage of fishery products at the temperatures laid down in Chapter VII. (iv) There needs to be somewhere for storing packaging materials and this is to be separate from product preparation and processing areas. where necessary. (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. recording of temperatures. (v) Special equipment for disposing of waste/ inedible fishery products into the sea or a watertight tank and allocation of separate areas. tanks must incorporate devices that allow a uniform temperature throughout the tanks. Requirements for factory vessels a) Factory vessels (i) These must have at a receiving area reserved for taking fishery products on board so each catch can be separated (ii) There must be a hygienic system for conveying fishery products from the receiving area to the work area (iii) Large work areas are needed to allow fishery products to be prepared and processed hygienically. and allow the monitoring and. to ensure that melt water does not stay in contact with the products. any waste processing unit needs to have a separate hold to store the waste. (iii) The above requirements for holds in vessels designed and equipped to preserve fresh fishery products for more than 24 hours must be met. Requirements for freezer vessels (i) These need to contain freezing equipment in order to achieve a core temperature of no more than -18 °C. A temperature recording device must be present and placed where the temperature in the hold is the highest.
Chapter II . 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.4. where fishery products are displayed for sale.Requirements during and after landing 9. and products washed thoroughly. (ii) In the case of food business operators responsible for auction and wholesale markets etc. B. b) Factory vessels on board which crustaceans and molluscs are cooked. Livers and roes for human consumption must be preserved under ice (temperature to approach that of melting ice) or be frozen.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. if this is not possible. 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.2. the product must be kept at no more than -9 °C. other than those kept alive. b) Fishery products are to be handled and stored in a way that prevents the flesh bruising. provided no damage occurs to the flesh. Parts that may present a danger to public health are to be removed quickly and kept away from products for human consumption. 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 . landed as soon as possible. chilled and wrapped. e) In cases where whole fish intended for canning is frozen in brine. The brine must not be a source of contamination.2 (i) Unloading and landing equipment that comes into contact with fishery products needs to be made of material that is easily cleanable. c) Factory vessels that freeze fishery products must have equipment meeting the requirements for freezer vessels as stated above.2. must be chilled or. d) Any gutting or heading of fish on board is to be done hygienically and quickly after capture. c) Fishery products. do not have to meet the above requirements if no other form of handling or processing takes place on board such vessels.
854/2004.Requirements for establishments. handling fishery products A. dispatched. 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. Additionally. Requirements for frozen products An establishment on land that freezes fishery products needs to have equipment that meets that for freezer vessels as stated in Section VIII.LEGISLATION consumption.2. B. (iv) Filleting and cutting is to be carried out in a way that avoids contamination or spoilage of fillets or slices. Fillets and slices must then be removed from work tables as soon as possible after preparation.4.2. They should be wrapped. Packaged fresh fishery products must be chilled to a temperature approaching that of melting ice. C. C. 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. part I. Requirements for fresh fishery products (i) Where chilled or unpackaged products are not distributed.3 Chapter III . prepared or processed immediately after arrival at an establishment on land. Requirements for mechanically separated fishery products (i) Raw materials used must satisfy the following 203 . 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. they must be stored under ice in appropriate facilities. packaged and chilled as quickly as possible after preparation. including vessels. (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. Chapter I. 9. (ii) Heading and gutting must be carried out hygienically. (iii) Whole and gutted fresh fishery products can be transported and stored in cooled water on board vessels. and where necessary.
and whole fish must be gutted and washed beforehand. (ii) mackerel. (iv) (wild) Atlantic and Pacific salmon. they must be cooled rapidly. if the processing is insufficient to destroy nematode larvae. Requirements for cooking of crustaceans and molluscs After crustaceans and molluscs are cooked.2.20 °C in all parts of the product for at least 24 hours (this treatment must be applied to the raw product or the finished product): (a) fishery products to be consumed raw or almost raw. Requirements concerning parasites (i) The fishery products below must be frozen at no more than . (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. or a stabilising treatment. (iii) sprat. Once produced. (iii) A document from the manufacturer. except when supplied to the final consumer.Requirements for certain processed fishery products A. cooling must continue until a temperature 204 . must accompany fishery products referred to above when placed on the market. and (c) marinated and/or salted fishery products.FISH AND SEAFOOD Only whole fish and bones after filleting may be used to produce mechanically separated fishery products.4 Chapter IV . stating the type of process they have undergone.4. If no other method of preservation is used. (ii) The manufacturing process must satisfy the following Mechanical separation is to be carried out immediately after filleting. mechanically separated fishery products must be frozen as quickly as possible or incorporated into a product intended for freezing. (b) fishery products from the following species. D. if they are to undergo a cold smoking process in which the internal temperature of the fishery product is not more than 60 °C: (i) herring. raw materials must have no guts.2. 9.
B. refining and purification steps before being placed on the market for the final consumer. centrifugation. B.4. Requirements for fish oil intended for human consumption (i) In relation to the raw materials used in their preparation and their transport. 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. and the raw material is processed within 36 hours after loading. However. cooked products are to be frozen or chilled to the temperatures stated in Chapter VII. the following conditions need to be met: A.Health standards for fishery products In addition to compliance with microbiological criteria laid down in Regulation (EC) No. 9. separation. (ii) The production process for fish oil must ensure that all raw material intended for the production of crude fish oil is subject to a treatment including (where relevant) heating.2. 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. 2074/2005).2. 205 . 2073/2005). raw materials need to be chilled as soon as possible and should meet the temperature requirements in Chapter VII. 2073/2005. 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. C. and after this. 2074/2005. Organoleptic properties of fishery products An organoleptic examination of fishery products must be carried out to ensure that fishery products comply with any freshness criteria (such criteria have yet to be set).LEGISLATION approaching that of melting ice is reached. pressing.5 Chapter V . Histamine Histamine limits must not be exceeded (limits are set in Regulation (EC) No. Shelling/shucking is to be done hygienically. processing.
E.FISH AND SEAFOOD D.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. Molidae. The scientific name must accompany the common name on the label. and cooked and chilled products from crustaceans and molluscs are to be kept at a temperature close to that of melting ice.6 Chapter VI .7 Chapter VII .2. 206 . In the case of whole frozen fish in brine intended for the manufacture of canned food. 9. prepared and processed fishery products belonging to the family Gempylidae. Toxins harmful to human health Fishery products derived from poisonous fish of the following families must not be placed on the market: Tetraodontidae. it should be kept at no more than -9 °C. 9. (ii) Frozen blocks prepared onboard vessels must be adequately wrapped before landing.2. Parasites Fishery products are to be visually examined so that visible parasites are identified before being placed on the market. 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. Diodontidae and Canthigasteridae. may only be placed on the market in wrapped/packaged form and must be appropriately labeled to provide information to the consumer on preparation/cooking methods and on the risk related to the presence of substances with adverse gastrointestinal effects. and when fishery products are wrapped on board fishing vessels.Storage of fishery products (i) Fresh fishery products. thawed unprocessed fishery products. (ii) Frozen fishery products must be kept at a temperature of not more than -18 °C in all parts of the product. in particular Ruvettus pretiosus and Lepidocybium flavobrunneum.2.4.2. Fishery products contaminated with parasites may not be placed on the market for human consumption. Fresh.4.
This covers Total volatile basic nitrogen (TVBN) limits and how to determine them.4.2. (ii) If fishery products are kept under ice.).8 Chapter VIII . 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. The temperature of frozen fishery products must not fluctuate above -15°C.4. Sebastichthys capensis. (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 . species belonging to the Merlucciidae family. 2074/2005. species belonging to the Gadidae family. (b) 30 mg of nitrogen/100 g of flesh for the species belonging to the Pleuronectidae family (with the exception of halibut: Hippoglossus spp. (c) 35 mg of nitrogen/100 g of flesh for the species: Salmo salar. Helicolenus dactylopterus. 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. 9.Transport of fishery products The above points for Chapter VII apply in relation to transport. The required temperature for whole frozen fish in brine intended for the manufacture of canned food during transport is not stated. provided that the journey is short and the competent authority allows this.3 Other information: Rules relating to visual inspections for the detection of parasites in fishery products are stated in Annex II (obligations on the competent authorities) of Regulation (EC) No. melt water must not remain in contact with the products. 9.2.LEGISLATION (iii) Fishery products kept alive are to be kept at a temperature which will not adversely affect food safety or its viability.2.
4. 9. tunicates and marine gastropods.FISH AND SEAFOOD Regulation (EC) No 853/2004. Member States may set limits at a higher level for certain species pending the establishment of specific Community legislation.3 The regulation details specific hygiene requirements for bivalve molluscs. they supplement the requirements of Annex II to that Regulation. Apart from the provisions on purification. however. Extracts of the requirements of Annex III. (b) and (c) of Part B(1) of that Chapter.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.1 Chapter I . Live bivalve mollusc requirements - 9. The sample must consist of about 100 g of flesh. (b) In the case of other operations. reference should be made to the actual regulation. specifically relating to bivalve molluscs are given below. they supplement the requirements of Annex I to that Regulation. The information required in the registration document is given in point 4 and includes the health status of the production area. where the raw material complies with points (a). 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. this Section also applies to live echinoderms. taken from at least three different points and mixed together by grinding. VI. A food business operator can only accept batch of such bivalve molluscs provided correct documentary requirements are presented.4. Chapters V. Chapter IX applies to pectinidae harvested outside those areas. VIII and IX. The copy of 208 . Section VII of Regulation 853/2004. The section is split into 9 chapters for which a summary is provided as follows: Chapters I to VIII apply to animals harvested from production areas that the competent authority has classified in accordance with Regulation (EC) No 854/2004. as amended. and point 3 of Chapter VII applies to retail. 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.3. please refer directly to the regulation. for full requirements. shellfish species etc.
Transport needs to allow sufficient drainage for best survival conditions. 209 . 9.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.4. Requirements for production areas (i) The competent authority classes (classed A. Other requirements such as conditions for relaying in order to allow optimal conditions for purification and for permanent record keeping are also specified. collection from: .2 Chapter II . (ii) Provided the requirements of chapter V are met. 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 A means they can be placed directly on the market . B.Hygiene requirements for the production and harvesting of live bivalve molluscs A.Class B means treatment in a purification centre or after relaying is done before it can be placed on the market .LEGISLATION document must be kept for at least 12 months post dispatch or receipt or longer as stated by the competent authority. 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. C.3. 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. poles or other fixed means are to identify the boundaries of the sites. processing or relaying. Requirements for relaying live bivalve molluscs Only areas approved by the competent authority can be used for relaying live bivalve molluscs with a requirement that buoys.
Requirements for purification centres There are requirements for live bivalve molluscs to be washed. 800 micrograms per kilogram. 1 milligram of yessotoxin equivalent per kilogram.Hygiene requirements for purification and dispatch centres A.Structural requirements for dispatch and purification centres Premises on land should not be subject to flooding by tides etc.4.3 Chapter III .4. and there are requirements for tanks and water storage containers. requirements if there are many batches of live bivalve molluscs. containers and a label to identify purification. Before dispatch. Requirements for dispatch centres Handling should not result in contamination to any degree.4 Chapter IV .3.5 Chapter V . the purification system to allow them to resume /maintain filter-feeding activity. B. (ii) amnesic shellfish poison (ASP). 210 . for: (i) paralytic shellfish poison (PSP). 160 micrograms of okadaic acid equivalents per kilogram.FISH AND SEAFOOD 9. the levels of marine biotoxins should not exceed the following limits.4. (iv) yessotoxins. dinophysistoxins and pectenotoxins together.Health standards for live bivalve molluscs In addition to microbiological criteria and organoleptic characteristic requirements.3. 20 milligrams of domoic acid per kilogram (iii) okadaic acid. 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. 9.3.
(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. 9.4.Identification marking and labelling In addition to general identification marking (contained in 9. by way of derogation from directive 2000/13/EC.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. they cannot be re-immersed in or sprayed with water. the label.1). 211 .Wrapping and packaging of live bivalve molluscs (i) Oysters must be wrapped or packaged with the concave shell downwards.3. must be waterproof.2. (Note: testing methods for detecting marine biotoxins are detailed in Annex III of Regulation (EC) 2074/2005) 9.3.8 Chapter VIII .4.7 Chapter VII .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. 160 micrograms of azaspiracid equivalents per kilogram.4. 9.LEGISLATION (v) azaspiracids. Registration document requirements also apply.9 Chapter IX .4. Also. The requirements for production areas as stated earlier apply to pectinidae and these can only be placed on the market through a fish auction. dispatch centre or processing establishment. including the identification mark. (ii) Once the live bivalve molluscs have been packed for retail sale and have left the dispatch centre. 9.3. the date of minimum durability may be replaced by the words ‘these animals must be alive when sold’. Finally.4. the species of bivalve mollusc and date of packaging (at least the date and month) are required.6 Chapter VI .3. Requirements of Chapters III and IV are to be complied with.
9. tunicates or marine gastropods containing marine biotoxins in total quantities exceeding the limits referred to in Regulation (EC) No 853/2004. or - - 212 .5. The official controls include audits of good hygiene practices and hazard analysis and critical control point (HACCP)-based procedures. chemical. 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. 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. The requirements refer to hygienic conditions and regular inspections both on land and at port to ensure compliance of temperature and other requirements. general principles for official controls in respect of all products of animal origin falling within the scope of the regulation are given. Chapter V of Regulation 853/2004 apply. or (iii) bivalve molluscs. Fishery products in particular must comply with the requirements of Annex III of the regulation.FISH AND SEAFOOD 9. In this regulation. 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. This refers to official controls or production and placing on the market of fishery products. physical or microbiological checks or checks for parasites have shown that they are not in compliance with the relevant Community legislation.5 Regulation (EC) No.1 Regulation (EC) 854/2004 Annex III Chapter III states that fishery products are to be declared unfit for human consumption if: organoleptic. they derive from: (i) poisonous fish. (ii) fishery products not complying with the requirement concerning biotoxins. echinoderms. Therefore the health standards for fishery products as previously stated in Annex III. Section VIII.
coli per 100 g of flesh and intravalvular liquid. Classification of production and relaying areas: This section reinforces that specified in Annex III. in 90% of the samples. 4. Annex II 9. The reference method for this analysis is the five-tube. as previously stated.5. chapter II on hygiene requirements for the production and harvesting of live bivalve molluscs in Regulation (EC) 853/2004. The reference method for this analysis is the five-tube. Also. - - - - B. coli per 100 g of flesh and intravalvular liquid.1 Chapter II .2 This states official controls for the production and placing on the market of live bivalve molluscs. Requirements for the competent authority are stated in relation to the classification or a production or relaying area is stated.000 E. Sampling plans and frequency are required. A summary is given as follows: 9. coli per 100 g of flesh and intravalvular liquid. Monitoring of classified relaying and production areas This is to ensure there is no malpractice. live bivalve molluscs must not exceed 46.Official controls concerning live bivalve molluscs from classified production areas A.5. three dilutions Most Probable Number (MPN) test specified in ISO 16649-3.2. Live bivalve molluscs from Class B areas must not exceed. that microbiological criteria and chemical contaminants amongst other requirements are met.600 E. live tunicates and live marine gastropods.LEGISLATION - the competent authority considers that they may constitute a risk to public or animal health or are for any other reason not suitable for human consumption. chapter V is reinforced. Live bivalve molluscs from class A areas may be collected for direct human consumption provided health standards are met. In the remaining 10 % of samples. section VII. live echinoderms. Live bivalve molluscs from Class C areas must not exceed 46. three dilutions MPN test specified in ISO 16649-3. Regulation (EC) 854/2004. 000 E. 213 .
FISH AND SEAFOOD C. 9.5.Official controls concerning pectinidae harvested outside classified production areas These are to be carried out in fish auctions. 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. Food business operators' own checks Controls that food business operators have done are considered by the competent authority when classifying.2 Chapter III . the competent authority is to close the production area. which has applied since 1 January 2006. 9. Procedures for re-opening a closed production area are stated D. 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. 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. E. 214 . 2073/2005.5. establishes microbiological criteria for a range of foods.6 Regulation (EC) No. 2073/2005 on Microbiological Criteria for Foodstuffs Regulation (EC) No. 9.3 Imports By way of derogation from Article 11(1) of Regulation (EC) No 854/2004. boundaries or class of a production area or its closure.2. dispatch centres and processing establishments in order to confirm that health standards and other specific requirements are met. 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. opening or closing a production area. F.
a batch of foodstuffs or a process.4 referring to fishery products.LEGISLATION The aim of this legislation is to complement food hygiene requirements. 9. per unit(s) of mass. and the legislation applies to all businesses involved in food production and handling. 215 . applying to products placed on the market and process hygiene criteria that are applied during the manufacturing process. and/or on the quantity of their toxins or metabolites.2 Process hygiene criteria Chapter 2 focuses on process hygiene criteria.II. The criteria are outlined in Table 9. area or batch. Two kinds of criteria have been established: food safety criteria. ensuring that foods being placed on the market do not pose a risk to human health.6.1 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. volume. 9. The relevant criteria are outlined in Table 9. presence or number of microorganisms. based on the absence. The definition of ‘microbiological criterion’ means a criterion defining the acceptability of a product.I. with chapter 2.6.
who has produced it EN/ISO 11290-2 (6) Products placed on the market during their shelf life 1. other than those intended for infants and for special medical purposes EN/ISO 11290-1 Products placed on the market during their shelf life Before the food has left the immediate control of the food business operator.16 Cooked crustaceans and molluscan shellfish Salmonella EN/ISO 6579 Products placed on the market during their shelf life . monocytogenes 1.1 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. other than those intended for infants and for special medical purposes(4) (8) L.3 Ready-to-eat foods unable to support the growth of L. monocytogenes 5 0 Absence in 25 g (7) FISH AND SEAFOOD 216 5 0 100 cfu/g 5 0 Absence in 25 g 1.2 Ready-to-eat foods able to support the growth of L.I Food safety criteria Sampling plan(1) n 10 5 Absence in 25 g EN/ISO 11290-1 c m M Products placed on the market during their shelf life Limit(2) Analytical reference method(3) Stage where the criterion applies Food category Microorganisms 1. monocytogenes L. monocytogenes.TABLE 9. monocytogenes.
manufactured from fish species associated with a high amount of histidine (17) Histamine . tunicates and gastropods 9 (18) 2 E.25 Live bivalve molluscs and live echinoderms. coli (15) 1.17 Live bivalve molluscs and live echinoderms.Food category Microorganisms Sampling plan(1) n 5 0 Absence in 25 g EN/ISO 6579 c m M Products placed on the market during their shelf life Products placed on the market during their shelf life Products placed on the market during their shelf life HPLC (19) Products placed on the market during their shelf life 400 g mg/kg Limit(2) Analytical reference method(3) Stage where the criterion applies 1.26 Fishery products from fish species associated with a high amount of histidine (17) 9 2 200 mg/kg Histamine LEGISLATION 217 1. tunicates and gastropods 1(16) 0 230 MPN/100 g of flesh and intravalvular liquid 100 mg/kg 200 mg/kg HPLC (19) ISO TS 16649-3 Salmonella 1.27 Fishery products which have undergone enzyme maturation treatment in brine.
spirits and similar products.FISH AND SEAFOOD 1 2 3 4 n = number of units comprising the sample. when he is not able to demonstrate. Engraulidae. 5 This criterion shall apply if the manufacturer is able to demonstrate. Valle M. Scombresosidae. Clupeidae. AOAC Internat. products with a shelf-life of less than five days shall be automatically considered to belong to this category. coli is used here as an indicator of faecal contamination. products with pH ≤ 5. Bouquelet S. E. live bivalve molluscs. Duflos G. References: 1. This criterion shall apply to products before they have left the immediate control of the producing food business operator. Assay of biogenic amines involved in fish decomposition. to the satisfaction of the competent authority. when recontamination is not possible after this treatment (for example. shall not apply. J. excluding sprouted seeds.4 or aw ≤ 0. Relevance of matrix effect in determination of biogenic amines in 6 7 8 15 16 17 18 19 218 . In such a case the presumption laid down in Article 14(6) of Regulation (EC) No 178/2002. Malle P. Other categories of products can also belong to this category. Bouquelet S.0 and aw ≤ 0. including cocoa and chocolate products. cider. m = M.. — fresh. Dervin C.25. wine. Products with pH ≤ 4. 1 ml of inoculum is plated on a Petri dish of 140 mm diameter or on three Petri dishes of 90 mm diameter. according to which the whole batch is to be deemed unsafe. 79. 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.. Single samples may be taken at retail level.. 43-9. Pomatomidae. The most recent edition of the standard shall be used.. Coryfenidae. 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. that the product will not exceed the limit 100 cfu/g throughout the shelf life.92. subject to scientific justification. — bread. Malle P. c = number of sample units giving values between m and M For point 1.94. beer. sugar. that the product will not exceed the limit of 100 cfu/g throughout the shelf life..1 . to the satisfaction of the competent authority. 1996. A pooled sample comprising a minimum of 10 individual animals. uncut and unprocessed vegetables and fruits. soft drinks. honey and confectionery. 2.1. biscuits and similar products. monocytogenes. Particularly fish species of the families: Scombridae. — — bottled or packed waters. products heat treated in their final package).
Salmonella in different food categories: — satisfactory. the mean value observed is ≤ m 2. if any of the values are > the limit. 82. 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. no values observed exceed the limit of M. J. tunicates and gastropods in relation to testing E. if all the values observed are ≤ the limit. if the following requirements are fulfilled: 1. if the presence of the bacterium is detected in any of the sample units.LEGISLATION plaice (Pleuronectes platessa) and whiting (Merlangus merlangus. where the limit refers to a pooled sample. monocytogenes in ready-to-eat foods able to support the growth of L. — unsatisfactory. coli. Interpretation of the test results relating to Table 9. if all the values observed indicate the absence of the bacterium. — unsatisfactory. a maximum of c/n values observed are between m and M 3. excluding live bivalve molluscs and live echinoderms. The test results demonstrate the microbiological quality of the batch tested(1).I The limits given refer to each sample unit tested. (1) 219 . if the presence of the bacterium is detected in any of the sample units. if the presence of the bacterium is detected in any of the sample units. — unsatisfactory. L. 1999. — unsatisfactory. monocytogenes before the food has left the immediate control of the producing food business operator when he is not able to demonstrate that the product will not exceed the limit of 100 cfu/g throughout the shelf life: — satisfactory. 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 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. L. 1097-101. monocytogenes in other ready-to-eat foods: — satisfactory. AOAC Internat. — unsatisfactory. monocytogenes in ready-to-eat foods intended for infants and for special medical purposes: — satisfactory. if all the values observed indicate the absence of the bacterium. L.
1 Shelled and E. coli 5 shucked products of cooked crustaceans and molluscan shellfish 2 100 cfu/g 1000 cfu/g EN/ISO 6888-1 or 2 FISH AND SEAFOOD Coagulasepositive staphylococci 5 Improvements in production hygiene 220 .TABLE 9.4.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.
— unsatisfactory. and the rest of the values observed are ≤ m.1 In the U. 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. The defendant may be required to produce well-founded scientific proof to support his claims. had not been 221 . — acceptable.K) Temperature control requirements 9. and particularly that perishable foodstuffs must not be kept at above the maximum recommended storage temperature. S. if one or more of the values observed are > M or more than c/n values are between m and M. but there is also a general requirement that foods must not be kept at temperatures that would result in a risk to health. if one or more of the values observed are > M or more than c/n values are between m and M. 2 Interpretation of the test results relating to Table 9. For example. tolerance periods for chill-holding of foods and hot-holding variations. The regulations prescribe a chilled food holding temperature of 8 °C or less.LEGISLATION 1 n = number of units comprising the sample. if a maximum of c/n values are between m and M. the defendant will need to prove that the food was on service or display. 2006 No. The regulations provide for defences in relation to upward variations of the 8 °C temperature.I.II The limits given refer to each sample unit tested. — acceptable. Coagulase-positive staphylococci in shelled and cooked crustaceans and molluscan shellfish: — satisfactory.K. if all the values observed are ≤ m. 2006 No.7 Food Hygiene (England) Regulations 2006. — unsatisfactory.I. with chill holding tolerance periods. The test results demonstrate the microbiological quality of the process tested. c = number of sample units giving values between m and M The most recent edition of the standard shall be used. 14 details temperature control requirements for foods in general. coli in shelled and shucked products of cooked crustaceans and molluscan shellfish: — satisfactory. S. 9. and the rest of the values observed are ≤ m. Schedule 4 of the Food Hygiene (England) Regulations 2006.7. E. which overrides the 8 °C requirement. if all the values observed are ≤ m. 14 (Hygiene requirements specific to the U. if a maximum of c/n values are between m and M.
was kept at above 8 °C or the recommended temperature for an unavoidable reason.K.FISH AND SEAFOOD previously put on display at more than 8 °C and had been kept there for less than four hours. 2006 No. 3).I. such as that below.S. 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. 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.8 Guidance In the U. for a limited period consistent with food safety. Schedule 4 of the Food Hygiene (England) Regulations 2006 contains several definitions. Alternatively. Recommended temperature: a specified temperature that has been recommended in accordance with a food business responsible for manufacturing. and was kept at above 8 °C or the recommended temperature. to or from premises (including vehicles) at which the food was to be kept at or below 8 °C or the recommended temperature.I. the period for which it can be expected to remain fit for sale when kept in a manner consistent with food safety. it would need to be proved that the food was being transferred to or from a vehicle used for the activities of a food business.uk/foodindustry/guidancenotes/hygguid/fhlguidance/ 222 . It should be noted that the temperature control requirements as detailed in Schedule 4 of the Food Hygiene (England) Regulations 2006 (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. 2006 No.food. or.gov. S. For other food. preparing or processing the food recommending that it be kept at or below a specified temperature between 8 ºC and ambient temperatures. or at a temperature above 63 °C and to reheat food to a temperature of at least 82 °C (The Food Hygiene (Scotland) Regulations 2006. The permitted reasons are given below: to facilitate handling during and after processing or preparation the defrosting of equipment. 9.
Regulation (EC) No. The revised text of Regulation (EC) No. 1791/2006 and 1021/2008 and as read with EC Directive 2004/41. 2076/2005.2005. L139. as read with the corrigenda at OJ No. 1791/2006 and 1020/2008 and as read with EC Directive 2004/41. 30. Regulation (EEC) 2136/89. 30. 55). 1688/2005. 62) as amended by Regulation (EC) No. The revised text of Regulation (EC) No.LEGISLATION 9. 854/2004 of the European Parliament and of the Council laying down specific rules for the organisation of official controls on products of animal origin intended for human consumption (OJ No. 2076/2005. Regulation (EC) No. Regulation (EC) No.2004. L338. 22. 25. L226. 1441/2007.9 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. 14.12.2004. 30. 2076/2005. 853/2004 is now set out in a Corrigendum (OJ No. 25. 25. 852/2004 of the European Parliament and of the Council on the hygiene of foodstuffs (OJ No.6.2004. laying down common marketing standards for preserved sardines. L226. Commission Regulation (EC) No. 2075/2005 and Regulation (EC) No.6. 2074/2005. 2074/2005 and Regulation (EC) No. Regulation (EC) No. 1019/2008 and as read with Regulation 2073/2005.2004. 854/2004 is now set out in a Corrigendum (OJ No. 1).2004. 1663/2006. 853/2004 of the European Parliament and of the Council laying down specific hygiene rules for food of animal origin (OJ No. 852/2004 is now set out in a Corrigendum (OJ No.2006. as amended. Regulation (EC) No. 22) as amended by Regulation (EC) Nos. L283. Regulation(EC) No. 223 . 83) as amended by Regulation Nos 882/2004. The revised text of Regulation (EC) No. 2074/2005. L226. 2076/2005.10. 206). these are not be discussed here. 1.4. L22.214.171.124 Further Reading Regulation (EC) No. 1662/2006. 3) as amended by Regulation (EC) No. 2073/2005 on microbiological criteria for foodstuffs (OJ No.4. 2074/2005. 9. and Regulation (EEC) 1536/92 laying down common marketing standards for preserved tuna and bonito. L139. namely.2004.
22.2001. (EC) No 854/2004 and (EC) No 882/2004 of the European Parliament and of the Council and amending Regulations (EC) No 853/2004 and (EC) No 854/2004 ( OJ L 338.12. 23.2005.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.12.1. as amended by Commission Regulation (EC) No 1022/2008 OJ L 338. 22. 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. 6) 224 . 83) Regulation (EC) No 104/2000 on the common organisation of the markets in fishery and aquaculture products (OJ L 17.2005.10. 21. 27 Commission Regulation (EC) No 2076/2005 laying down transitional arrangements for the implementation of Regulations (EC) No 853/2004.2000. 22) Commission regulation (EC) No 2065/2001 laying down detailed rules for the application of Council Regulation (EC) No 104/2000 as regards informing consumers about fishery and aquaculture products as amended by Commission Regulation (EC) No 1792/2006 of 23 October 2006 (OJ L 278.
1. respectively (1).3.2 Oxygen requirements Facultative anaerobe.45 °C (1).5 µm 10.3 .1 Aeromonas spp.4 Heat resistance Aeromonads are sensitive to moderately high temperatures.10.3 Temperature Aeromonas grows at temperatures of 2 . under optimal incubation temperatures.0 .5. 10.000-fold increase in number is observed after one week in food systems (1).1. Some death will occur at this pH if the temperature is 4 °C. Aeromonas has the potential to grow over a pH range of ca 4 .0 x 1. 0. At 5 °C. Morphology Gram-negative rods. and D-values at 45 and 51 °C have been reported as 29.5 pH Under otherwise optimal conditions. The optimum growth temperature is 28 °C (2).1. PATHOGEN PROFILES 10. and 2. 10. 10. The minimum pH value for growth is thought to be pH 5. a 10 to 1.1.1 10.5.1. 225 . no growth is observed (1).10 (3).5 minutes.3 minutes. At pH 4.1.
being capable of slow growth and toxin production at low temperatures .2. 226 . types B and F can be either. botulinum is prevented.even as low as 3 ºC (7).0 µm.6 Aw/Sodium chloride Growth of Aeromonas is optimal in the presence of 1 . which may be inactivated by heating at 80 ºC for 20 . botulinum are not particularly heat resistant. A to G. F are the types that are responsible for most cases of human botulism (5. Types A.2.4 x 1.22.2 Oxygen requirements Although C. forming at least seven different toxins. but the spores of this organism are more so.5 . 10.2 Clostridium botulinum Seven different types of C. spore-forming rods. many foods that are not obviously ‘anaerobic’ can provide adequate conditions for growth.2% sodium chloride (NaCl) (1).2. 10. 10. to a lesser extent. botulinum grow reasonably well in the temperature range of 20 45 ºC. Group I will not grow at temperatures of 10 ºC or less. 0. 6).2. There are four main groupings of the organism. 10.1. All type A strains are proteolytic and type E strains are usually non-proteolytic. and is sensitive to > 4. or at 90 ºC for a few seconds. but Group II strains are psychrotrophic. Thus an aerobically packed product may not support the growth of the organism on the surface. It is also important to note that the inclusion of oxygen as a packaging gas cannot ensure that growth of C. but the interior of the food may do so. E and. at 85 ºC for 5 minutes.5% NaCl (4).3 Temperature All strains of C.7 .1 Morphology Gram-positive.4 Heat resistance The vegetative cells of C. B.30 minutes. 10. but the minimum temperatures required to inhibit Groups I (proteolytic group) and II (non-proteolytic group) are different. botulinum are known.FISH AND SEAFOOD 10.2. botulinum types produce heat labile toxins. botulinum is an obligate anaerobe. and groups I and II are those responsible for the botulism cases. All C.
1.0.2 Oxygen requirements C.2. the minimum aw for growth may be lower.2. but under optimum growth conditions 10% (w/w) NaCl is required to prevent growth of Group I. However. botulinum spores The most heat resistant spores of Group I C. vacuum packed. botulinum depends on solute.10.3 10.is an obligate anaerobe. pH and temperature. 0.2.3. 10. parcooked meals with pH value > 5.g. 10. 11)). perfringens .85 ºC) and their ability to grow at refrigeration temperatures necessitates their control in foods capable of supporting their growth (e. therefore. and 5% (w/w) NaCl is necessary to prevent growth of Group II organisms.94 for Group I and 0.0 µm.97 for Group II (5).0 . botulinum are produced by type A and proteolytic B strains (D121ºC=ca 0. grow on the surface of foods unless they are vacuum or gas-packed. 10.3 . 10. The spores of Group II (non-proteolytic/psychotrophic) strains are less heat resistant than Group I strains. safety margins must be introduced to allow for process variability.5 pH The minimum pH for the growth of proteolytic and non-proteolytic strains is pH 4.21 minutes) (9). 227 . In commercial situations. they may survive mild heat treatments (70 .6 and pH 5.7 Characteristics of C.6 Aw/ Sodium chloride The minimum water activity (aw) for growth of C.1 Clostridium perfringens Morphology Gram-positive spore-forming rods. particularly by cooking. where oxygen levels are reduced. Note also that if humectants other than salt are used. 9). The organism will grow well in the centre of meat or poultry dishes.97) (10.PATHOGEN PROFILES 10. These values have been established under carefully controlled laboratory conditions. It will not. These concentrations correspond to limiting aw of 0.like other clostridia .3.0 and aw > 0.9 x 2. respectively (8.
While some growth can occur at 50 ºC. very limited growth may occur at pH values over the range pH ≤ 5 and ≥ 8. perfringens. The lowest aw recorded to support the growth of C. perfringens in vacuum packed beef (18). Salt concentrations of 6 . death of the vegetative cells of this organism usually occurs rapidly above this temperature (12. At cold temperatures. 10. 16). perfringens is not a tolerant organism with respect to pH. depending on strain and other conditions. perfringens in roast beef at 60 ºC is 14.6 Aw/Sodium chloride C.4 Heat resistance Exposure to a temperature of 60 ºC or more will result in the death of vegetative cells of C. perfringens has the potential ability to grow within the temperature range 15 . the enterotoxin is not heat resistant . 10. will survive greater extremes of pH (and aw) (12.93 and 0. pH.50 ºC.97 (glycerol and sucrose respectively) depending on the solute used to control aw (13. although prior growth at high temperatures or the presence of fat in a food will result in increased heat resistance. As in the case of other factors limiting the growth/survival of this organism. perfringens is not tolerant of low water activities.5 pH C. 13).5 minutes (17).3. Its optimum temperature for growth is 43 . vegetative cells die rapidly (12). 13). although C.7 Characteristics of C. the limits for aw are affected by temperature.3. Under otherwise ideal conditions.8% inhibit growth of most C. In addition. however. Spores.3. (It is unusual for spores to be formed in foods after the growth of this organism (14)).45 ºC. Some studies indicate that the presence of 3% NaCl delays growth of C.3 Temperature The most significant characteristic of C. perfringens strains. type of solute.3.3. It grows best at pH values between 6 and 7 (the same pH as most meats). and their heat resistance is also affected by the heating substrate. 18). perfringens can vary quite considerably in their heat resistance. perfringens appears to be 0.10 ºC.it is destroyed by heating at 60 ºC for 10 minutes (14.FISH AND SEAFOOD 10. lower concentrations may be effective in combination with other factors. 10. etc. 10. The D-value for C. perfringens in relation to food safety is the organism’s ability to grow extremely rapidly at high temperatures. Recorded heat 228 .3. 0 . 15. perfringens spores The spores of C.
0 minutes for heat resistant spores to 1.4. 0. 229 . 60 and 65 ºC were reported as 10. 10. being potentially capable of growing . pH < 5.4 Heat resistance L.4 ºC is probably a more likely minimum for growth in foods (19). is between 30 and 37 ºC.0 µm. The upper temperature limit for the growth of L.4 10.23. the lowest limit for growth is likely to be considerably higher .8 minutes for heat sensitive spores (12). 10.5 x 0.2 has been suggested as the lowest working limit (22).4.at refrigeration temperatures down to. using hydrochloric acid as an acidulant).4. monocytogenes is not a particularly heat resistant organism.2.19 minutes. Under ideal conditions. D-values for L. However. and temperature. monocytogenes is reported to be 45 ºC (20).especially at refrigeration temperatures.64.6 . so can be destroyed by pasteurisation. and where acetic acid is used as an acidulant.albeit slowly .98 and 0.2 Oxygen requirements Aerobe or microaerophilic. 10. 1.5 . It has been reported to have slightly greater heat resistance than certain other foodborne pathogens.PATHOGEN PROFILES resistance values at 95 ºC (D-values) range from 17.3 Temperature Listeria monocytogenes is unusual amongst foodborne pathogens in that it is psychrotrophic. growth at low temperatures can be very slow. respectively (21). Its optimum growth temperature. 10.3 is the lowest value where growth has been recorded. or even below 0 ºC.0.5 pH The ability of Listeria to grow at different pH values (as with other bacteria) is markedly affected by the type of acid used.4. -0.3 . monocytogenes in crawfish tail meat at 55.1 Listeria spp. 10.4 . Morphology Gram-positive short rods.2. requiring days or weeks to reach maximum numbers. it is not a sporeformer. the organism is able to grow at pH values well below pH 5 (pH 4.4. In foods. however. however.
83 (20). 10.8 . 10.1. It grows best at aw of ≥ 0.0 µm.6 Aw/Sodium chloride L.5. Morphology Gram-negative short rods.5 .5.45 ºC.6 10.0 .0.5 .5. monocytogenes is quite tolerant of high NaCl/low water activities.1 Salmonella spp.FISH AND SEAFOOD 10.5. peritrichous flagella.97.0 x 1 . 25). or even grow.4 Heat resistance Pasteurisation at 60 ºC for 30 minutes has been reported as being effective in killing Plesiomonas shigelloides (24).4. 10. It is likely to survive.7 x 1.8.3. 0. 10.6 Aw/Sodium chloride Plesiomonas can grow well at NaCl concentrations up to 4% (24. 0.1 Plesiomonas Morphology Gram-negative.90. The bacterium may survive for long periods at aw values as low as 0. 10. the optimum being 30 ºC (23).12% NaCl or more). at salt levels found in foods (10 . 10.5 10.5.5 pH Plesiomonas has a pH range for growth of pH 4. but has been shown to be able to grow at an aw level of 0. 25).3 Temperature Plesiomonas can grow in the temperature range of 8 .5.6.2 Oxygen requirements Facultative anaerobe.3 µm. 10.5 (24. 230 .
4% NaCl. However.4 Heat resistance Salmonella is not a spore-forming organism. depending on serotype. depending on the type of acid and temperature. Salmonellae are quite resistant to drying. although salt tolerance increases with increasing temperature (28). and appropriate heat treatments must be determined experimentally for low-aw foods. under otherwise ideal conditions.37 ºC.8 . with a z-value of 4 .PATHOGEN PROFILES 10.5 ºC.6 Aw/Sodium chloride Where all other conditions are favourable.0.2 Oxygen requirements Facultative anaerobe. and with an appropriate acid.945. substrate. However.20 times more heat resistant than the average strain of Salmonella at high aw (27).6. the optimum being 35 . Salmonella has the potential to grow at aw levels as low as 0.5.48 ºC. 231 .93 (as reported in dried meat and dehydrated soup).6.5. It is not. some strains are able to grow at temperatures as low as 4 °C (26). Some death will occur at pH values of less than about 4. 10. a heat resistant organism.6. Salmonellae are quite resistant to freezing. The growth of Salmonella is generally inhibited by the presence of 3 .5 . D60 ºC values normally range from about 1 to 10 minutes. 10. Furthermore. The optimal pH for Salmonella growth is between 6. or possibly 0. pasteurisation and equivalent treatments will destroy the organism under normal circumstances. 10. temperature and pH. Growth is slow at temperatures below about 10 ºC.5 pH Salmonella has a pH range for growth of pH 3. high fat or low moisture levels (low aw) will reduce the effectiveness of heat treatments. therefore.6. 10.9.3 Temperature Salmonellae can grow in the temperature range of 7 . Salmonella senftenberg 775W is about 10 .7.6. strains vary in their ability to withstand heating.
7. the D60 ºC value is 2 .7.7 10. aw and temperature (sensitivity to acid increases with temperature). 10.0. 10.2 Oxygen requirements Facultative anaerobe. with an optimum of 35 . Staph. 10. Freezing and thawing have little effect on Staph.4 Heat resistance Heat resistance depends very much on the food type in which the organism is being heated (conditions relating to pH. The limits for toxin production are also narrower than for growth. In meat macerate. but may cause some cell damage (30). stressed cells can also be less tolerant of heating. 0.5 ºC. aureus viability.FISH AND SEAFOOD 10.7.7.9 µm diameter. the organism is heat sensitive and will be destroyed by pasteurisation.). As is the case with other bacteria.2 to 9.3 (optimum 7.7. aureus can grow within the temperature range 7 – 48. aureus is more limited under anaerobic than aerobic conditions. aw. fat content. Most strains of staphylococci can grow within the pH range 4. Under most circumstances.37 ºC (29).3 Temperature Under otherwise ideal conditions. etc. NB: The growth of Staph. aureus than citric acid). 10. It can survive well at low temperatures. under otherwise ideal conditions (29. 31). however. 232 .7.5).20 minutes.0 .1 Staphylococcus aureus Morphology Gram-positive cocci.5 pH The pH at which a staphylococcal strain will grow is dependent upon the type of acid (acetic acid is more effective at destroying Staph.7 . The following relate to limits for growth only. depending on aw.
>0. 31) between 0.0. an aw of 0.3 Temperature The optimum temperature for growth has been reported as 30 ºC.7 Limits permitting toxin production 10 .87 and > 0.8. Staphylococci are more resistant to salt present in foods than other organisms. An effect of increasing salt concentration is to raise the minimum pH of growth.10% NaCl. strain.1 Vibrio cholerae Morphology Gram-negative curved rods.4 .8. atmosphere.99 aerobically under otherwise optimal conditions. 0.7.2. pH. 233 .0 . * dependent on temperature.PATHOGEN PROFILES 10.7.9. and solute.83 . In general.46 ºC (optimum between 35 and 40 ºC) (very little toxin is produced at the upper and lower extremes) (31) 5. but certain strains can grow in 20% NaCl. especially vacuum packed foods (33) Temperature: pH: *Aw: Atmosphere: Heat Resistance: Enterotoxins are quite heat resistant.8 x 1. In general. aureus can grow in 7 . 10. It can grow over the aw range 0.43 ºC (35). heating at 100 °C for at least 30 minutes may be required to destroy unpurified toxin (31. 10.99 little or no toxin production in anaerobically packed foods.8. 34).86 is the generally recognised minimum in foods (32).8 10.5) (29.6 Aw/Sodium chloride Staph. 10.0 (optimum 7. However. with a temperature range for growth of 10 .6 µm 10. Staph.7.2 Oxygen requirements Facultative anaerobe.2 .5 . aureus is unusual amongst food-poisoning organisms in its ability to tolerate low water activities.
6. under otherwise favourable conditions.9 10. cholerae is sensitive to heat at temperatures of >45 ºC. 234 . but only moderately sensitive to freezing (40).97 .6 Aw/Sodium chloride V.4.984 (35). 38). and it is sensitive to refrigeration temperatures (0 .0% NaCl (35). it has a pH range for growth of pH 5 . declining in number during storage.5 pH This organism is tolerant of high pH values. cholerae may persist on frozen fish for >180 days (36). with optimum at 7. parahaemolyticus is easily destroyed by drying.3 Temperature The minimum and maximum temperatures reported for growth in laboratory media. 37. It can grow in the presence of 0. are 5 and 43 ºC. 10. 10.65 minutes (35).1 .9.5 ºC). cholerae in crab meat homogenate at 60 ºC is 2. 10. respectively. with an optimum of 37 ºC (39).9. V.9. but not acid conditions.8. with an optimum of 0.998.8.2 Oxygen requirements Facultative anaerobe.8.1 Vibrio parahaemolyticus Morphology Gram-negative curved rods. The D-value for V. 10.0.FISH AND SEAFOOD Also. cholerae can grow over the aw range 0. 10.6 (35. there is evidence that V.9. 10.4 Heat resistance V.
and are readily destroyed by cooking.9.6 .4 Heat resistance V.PATHOGEN PROFILES 10.4 Heat resistance V.6 Aw/Sodium chloride V.0.5 pH The pH range for growth is 4. respectively (39).5% NaCl. The D-values at 55 ºC for clam and crab homogenate are 0.4% (39. 235 . The optimum concentration of NaCl equates to aw of approximately 0.2 Oxygen requirements Facultative anaerobe. parahaemolyticus will grow at aw levels as low as 0. 41). 10. it has been reported that cooking oysters for 10 minutes at 50 ºC should ensure destruction of the organism (43). but the optimum concentration is 2 .8. 10.43 ºC (42).10. and 2.9.02 .10.9.8 . 10.11.10. parahaemolyticus is unable to grow in the absence of salt.10. vulnificus is heat sensitive. depending on solute. V. 41). The organism can survive for long periods at freezer temperatures (42).10 Vibrio vulnificus 10. 10. respectively.3 Temperature The optimum temperature for growth has been reported at 37 ºC.6 (41). with an optimum pH of 7. 10. 10.29 minutes. with a temperature range for growth of 8 .1 Morphology Gram-negative curved rods.922.5 minutes. its upper and lower limits for growth are 8 (although some reports suggest 10) and 0.98 (35. parahaemolyticus are not heat resistant.
Ed.L. Caister Academic Press.5.W. Marcel Dekker. 2005. Cliver D.M. Eds.W.. Dodds K. 6. Diseases Caused by Bacteria. in Food Microbiology: Fundamentals and Frontiers. Marcel Dekker.. 1995.FISH AND SEAFOOD 10. in Microorganisms in Foods. Notermans S. New York. ASM Press. Eds.10.10. in Foodborne Disease Handbook. Palumbo S. Kim J. 1-19.5 . Volume 5: Microbiological Specifications of Food Pathogens. 111-89.N.J. Dodds K.K. in Foodborne Bacterial Pathogens..O.. Bhunia A..0% NaCl.R. New York.D.. Marcel Dekker Inc. New York. with an optimum pH of 7.C. in Foodborne Pathogens.W.W. Ed. Aeromonas species. Doyle M. Marcel Dekker. International Commission on Microbiological Specifications for Foods. 10.L. 236 . Great Britain. 1989.H. in Foodborne Disease Handbook. Stelma G.H. Fratamico P. Kovacek K.D. Growth and heat resistance of psychrotrophic Clostridium botulinum in relation to ‘sous vide’.R.M.. Murrell K.L. Volume 1. Marcel Dekker. Clostridium botulinum.L. with an optimum of 2. Microbiology and Molecular Biology.. 329-49. 357-67.10.. Clostridium botulinum.. 2. 1994... 11. Gorman J. Hauschild A.. 1989.8 (42). Eds. Aeromonas hydrophila.. Eds. Doyle M. Volume 1: Diseases Caused by Bacteria. 1993.P. Blackie.. Aeromonas hydrophila group. 10.H. Ed. Clostridium botulinum and Clostridium perfringens. Washington D.E. 3. London. Bier J. 6 (1). 1-27. 383-408. Eds.. Smith J. 1993. Miliotis M. 8.N.A. Montville T. Cliver D. Food Control. in Clostridium botulinum: Ecology and Control in Foods. Betts G. 279-303. Austin J.H.H. References International Commission on Microbiological Specifications for Foods.P.. 9.. Faris A. 1994.. Marcel Dekker. Gorham J.P. Clostridium botulinum. Gaze J. vulnificus grows over the range 0. Stelma G. Dodds K.. Hauschild A. Peck M.. Doyle M. Johnson E.R. New York. Abeyta C. 57-63.6 Aw/Sodium chloride V. 2003. New York. 1996. Principles of control. Murrell K. New York.H. Marcel Dekker. Potential hazards associated with REPFEDS. 5-19. 10. Lund B. 121-76. Eds. Juneja V.D. 5. 7. Aeromonas. Foegeding P. 2001.5% (42)..M.5 pH The pH range for growth is 5 .W. Beuchat L. Hauschild A. in Foodborne Bacterial Pathogens.O.11 1. 4. in Clostridium botulinum: Ecology and Control in Foods. Hui Y.. Novak J. Hui Y. Eds. in International Handbook of Foodborne Pathogens. 97-131.D. New York.
Clostridium perfringens. 1996.C.. Doyle M. 26. 2005.. Montville T.. Clostridium perfringens.P.K. 237 .A. 1990. Journal of Applied Bacteriology.A. 18. Volume 1. in Microorganisms in Foods. Emery D. International Commission on Microbiological Specifications for Foods. Elsevier. Labbe R. New York. in Food Microbiology .P. International Commission on Microbiological Specifications for Foods.M.M. Juneja V.PATHOGEN PROFILES 12. Ed.R. Marcel Dekker. Volume 5: Microbiological Specifications of Food Pathogens.A. 2001. London.R. 19. Doyle M. in Foodborne Pathogens: An Illustrated Text. 14.R. Cliver D. 15. Murrell K. Varnam A. Effect of time and temperature on growth of Salmonella enteritidis in experimentally inoculated eggs. Blackie. 1994. International Commission on Microbiological Specifications for Foods.P. Evans M.V..G. 1990. 2007. Gorham J. Lancet. Eds. Koburger J.D.H. Johnson E. Avian Disease. Plesiomonas. Riemann H. 1996.. New York. London. Wolfe Publishing Ltd.P. Marcel Dekker. Plesiomonas shigelloides. Blackie. Plesiomonas shigelloides. Ed. Listeria monocytogenes. Growth of Listeria monocytogenes at refrigeration temperatures. International Commission on Microbiological Specifications for Foods. 1989.. 21.An Introduction. Washington D.. Archer P. 17.. Clostridium perfringens food poisoning. in Microorganisms in Foods. Marcel Dekker.T. Ed. 735-42..G. in Foodborne Infection and Intoxication. McClane B. London.H. Marcel Dekker.A. Banks J..A. 23. 133-67. in Foodborne Disease Handbook. Diseases Caused by Bacteria. 351-82. Listeriosis and Food Safety. 311-25. 112-25. ASM Press. Matthews K. Blackie. London. Academic Press.. Lund B. Walker S.J. in Foodborne Diseases. in Microorganisms in Foods. London. Clostridium perfringens. Ed. 68 (2). Cliver D. New York. Eds.J. in Foodborne Bacterial Pathogens. Ryser E. Cliver D. 33. Marth E. 191-243. Clostridium perfringens. Clostridium perfringens gastroenteritis. 20. 22. Listeria monocytogenes. Foodborne disease due to Bacillus and Clostridium species.. 208-13. Hui Y. Evans M.O. 336 (8721).C.. Labbe R. Kim C.. Eds. 229-40. Eds. 1990. Listeria.O. London. 982-6.H.. Beuchat L. Halvorson D. 1989. Rinke H. International Commission on Microbiological Specifications for Foods. in Foodborne Bacterial Pathogens. 201-8.H. 2006.G..J. 1991. in Food Microbiology: Fundamentals and Frontiers. Volume 5: Microbiological Specifications of Food Pathogens.. 1989. 1996. Ed. 137-64. 16. New York.. Ed. Volume 5: Microbiological Specifications of Food Pathogens. 13. 157-62. Varnam A. Nagaraja K. Doyle M. 129-38. ASM Press. Eds. Wrigley D.O. International Commission on Microbiological Specifications for Foods. 141-82. Washington D. Montville T. 24.J. 25.
2005.M.S. Staphylococcus aureus. in Foodborne Pathogens: Microbiology and Molecular Biology.. in Microorganisms in Foods. Oliver J. 40. D’Aoust J-Y. Salmonella. Wymondham.62. Ed. 33. 543-68. Food and Environmental Sanitation.M. Caister Academic Press. Ed. International Commission on Microbiological Specifications for Foods. Golden D.P. 1990. 1989. Staphylococcal Food Poisoning. Smith J. Blackie.A. 1930. Volume 5: Microbiological Specifications of Food Pathogens.P. Salmonellae. Vibrio cholerae. in Foodborne Disease. International Commission on Microbiological Specifications for Foods. Bergdoll M. 642. Staphylococcal gastroenteritis.O. 28. in Microorganisms in Foods.H.P. 2005. 545-66. CDR Review.. Woodhead Publishing Ltd. London. 38. Blackie.J. 1993. Jay J.. Wilkinson B. Microbiological Specifications of Food Pathogens. 2005. Microbiological Specifications of Food Pathogens. 45 (3). 35.. Eds. International Commission on Microbiological Specifications for Foods. Volume 5.R. 327-52.. Springer Science. 34. 1983.A. Cliver D. 985-98. London. 85-106. Japanese Journal of Experimental Medicine.FISH AND SEAFOOD 27.O. Ed. International Commission on Microbiological Specifications for Foods.A. Doyle M. Ed.S. 331-57. in Microorganisms in Foods. New York. 426-35.M. Volume 5.S. London. 1996. Cleland D. 217-64.. 1989. in Foodborne Bacterial Pathogens. Dairy. Warner R. London. 2005. Vibrio parahaemolyticus. London. Twedt R. 1994. Gustafson J. Ed.C.J. R15764. Golden D.M. Staphylococcus aureus as a food pathogen: staphylococcal enterotoxins and stress response systems. Foodborne illness (Part 1): Staphylococcal (“Staph”) food poisoning. Ed. in Modern Food Microbiology. Ed. 29. Griffiths M. in Foodborne Pathogens. 523.. 273-84. Bergdoll M. in Foodborne Infections and Intoxications. Stewart G. Lee-Wong A. 4 (13). Marcel Dekker. Doyle M. 32. Vibrio parahaemolyticus. 13 (11). Cholera: current epidemiology. Loessner M. New York. Cliver D. Marcel Dekker. 1996. Academic Press. Fratamico P. Bhunia A.. International Commission on Microbiological Specifications for Foods. 36. Tohyama Y. Loessner M. Cambridge. 414-25. Applied and Environmental Microbiology. Distribution of Vibrio vulnificus and other lactose-fermenting vibrios in the marine environment.L. Reed G. Stress Response and Resistance. Riemann H. Academic Press. 238 . Eds. 1996. 8.C. in Understanding Pathogen Behaviour Virulence. Eds. Blackie. International Commission on Microbiological Specifications for Foods. 39.K. New York. 31. 30. 37. 327-445. Jay J. Staphylococcal intoxications. Experimental investigations concerning the prevention of epidemic diseases due to aquatic products (Fourth Report).D. Crowcroft N... On the resisting power of Vibrio cholerae and typhoid Bacillus against low temperatures.
and Campylobacter species. 55 (12). 436-9. Ed. 239 .. 1996..D.W. Golden D. in Microorganisms in Foods. Cook D. Vibrio vulnificus. International Commission on Microbiological Specifications for Foods. Cold storage and mild heat treatment as processing aids to reduce the numbers of Vibrio vulnificus in raw oysters. Journal of Food Protection.J.A. 657-78.A. Loessner M. Ruple A. 985-9. New York. 43. 1992. 42.. Microbiological Specifications of Food Pathogens. Yersinia.J. Volume 5. 2005. Jay J. in Modern Food Microbiology. Springer Science.PATHOGEN PROFILES 41.M. Blackie. International Commission on Microbiological Specifications for Foods. London. Golden D... Jay J. Loessner M.M. Foodborne gastroenteritis caused by Vibrio.
FISH AND SEAFOOD 240 .
uk/Topics/Fisheries Seafish Industry Authority 18 Logie Mill Logie Green Road Edinburgh EH7 4HS United Kingdom Tel: + 44 (0)131 5583331 Fax: + 44 (0)131 5581442 Web site: www.CONTACTS Addresses of Trade Associations and Professional Bodies Association of British Salted Fish Curers and Exporters c/o Cawoods (Fish Curers) Ltd Estate Rd No 6 South Humberside Industrial Estate Grimsby North East Lincolnshire DN31 2TG United Kingdom Tel: + 44 (0) 1472 342248 Fax: + 44 (0) 1472 353100 Email: firstname.lastname@example.org/ .scotland.gov.defra.uk Web site: www.gov.seafish.ifm.uk/ Department for Environment Food and Rural Affairs (Defra) Nobel House 17 Smith Square London SW1P 3JR United Kingdom Tel: + 44 (0) 207 2386000 Fax: + 44 (0) 207 2382188 241 Email: email@example.com@ifm.uk Web site: www.uk Prepared Fish Products Association (PFPA) Federation House 6 Catherine Street London WC2B 5JJ United Kingdom Tel: + 44 (0) 208 78362460 The Scottish Government Fisheries Group Environment and Rural Affairs Department Pentland House 47 Robb's Loan Edinburgh EH14 1TY United Kingdom Tel: + 44 (0) 8457 741741 or + 44 (0)131 5568400 Fax: + 44 (0)1397 795001 Email: firstname.lastname@example.org Web site: www.uk or v.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.
gov.no Web site: www.uk/index/fisheriesfarming-and-food.gov.no/ Matís ohf (Icelandic Fisheries Laboratories) Skúlagata 4 101 Reykjavík Iceland Tel: + 354 422 5000 Fax: + 354 422 5001 Email: firstname.lastname@example.org.C.ie/templates/homepage.eapo.com/ Danish Institute for Fisheries Technology and Aquaculture (DIFTA) illemoesvej The North Sea Centre DK-9850 Hirtshals Denmark Tel: + 45 98 944300 Fax: + 45 98 942226 Email: email@example.com. Dublin Eire Ireland Tel: + 353 1 2144100 Web site: www.13 Postbox 6122 N-9291 Tromsø Norway Tel: + 47 77 629000 Fax: + 47 77 629100 Email: post@fiskeriforskning. Andersens Boulevard 37.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.fiskeriforskning. 1 1553 København V Denmark Tel: + 45 35 372023 Fax: + 45 35 372033 Email: Info@fiskehandlerne. Northern Ireland Fisheries and Rural Policy Division 4th Floor North Dundonald House Stormont Estate Belfast BT4 3SB United Kingdom Tel: + 44 (0) 28 90524999/ 4991 Fax: + 44 (0) 28 90378323 Email: firstname.lastname@example.org. Baelskaai 25 8400 Oostende Belgium Tel: + 32 59321876 Fax: + 32 59322840 Email: email@example.com/fishery/ European Association of Fish Producers’ Organisations H.org Web site: www.uk Web site: www.com Web site: www.is Web site: www.matis.net Irish Sea Fisheries Board PO Box 12 Crofton Road Dun Laoghaire Co.ifpea@eircom.FISH AND SEAFOOD Department of Agriculture and Rural Development.dk Danmarks Fiskehandlere H.fiskehandlerne.dk Web site: www.dk/s Norwegian Institute of Fisheries and Aquaculture Ltd Muninbakken 9 .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.dk Web site: www.is/ 242 .
uk Web site: www.org.ifst.fdf.nz Web site: www.CONTACTS Fishing Industry Research Institute 15 Lower Hope Street Rosebank 7700 Republic of South Africa National Fisheries Institute (USA) 7918 Jones Branch Drive Suite 700 McLean VA 22102 United States of America Tel: + 1 703 7528880 Email: firstname.lastname@example.org/en/index.uk Department of Health Richmond House 79 Whitehall London SW1A 2NS United Kingdom Tel: + 44 (0) 207 2104850 Email: email@example.com New Zealand Seafood Industry Council Private Bag 24-901 Wellington 6142 New Zealand Tel: + 64 4 3854005 Fax: + 64 4 3852727 Email: firstname.lastname@example.org Web site: www.com/ 243 .uk Web site: www.dh.csiro.com Web site: www.uk Institute of Food Science and Technology (IFST) 5 Cambridge Court 210 Shepherds Bush Road London W6 7NL United Kingdom TeI: + 44 (0) 207 6036316 Fax: + 44 (0) 207 6029936 Email: email@example.com 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 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.au 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.co.cmar.co.gsi.org.bsi-global.org CSIRO Marine Laboratories Castray Esplanade GPO Box 1538 Hobart Tasmania 7001 Australia Tel: + 61 3 62325222 Fax: + 61 3 62325000 Email: email@example.com 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: cservices@bsigroup.
uk Web site: www.1211 Geneva 27 Switzerland Tel: + 41 22 7912111 Fax: + 41 22 7913111 Email: firstname.lastname@example.org/web/indexen.int Web site: http://www.00187 Rome Italy Tel: + 39 06 487751 Fax: + 39 06 4877599 Email: email@example.com/foodsafety World Health Organization (WHO) Headquarters CH .org.euro.hps.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: 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.euro.int Web site: www.int/ SIK .de 244 .sofht.who.codexalimentarius. jsp Food Safety 10 .scot.uk Web site: www.int/en/ WHO Regional Office for Europe Scherfigsvej 8 DK-2100 Copenhagen Ø Denmark Tel: + 45 39 171717 Fax: + 45 39 171818 Email: email@example.com/ Health Protection Scotland Clifton House Clifton Place Glasgow G3 7LN United Kingdom Tel: + 44 (0)141 3001100 Fax: + 44 (0)141 3001170 Email: hpsenquiries@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.uk/index.who.who.int.sfam.who.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/scieh.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: admin@sofht. Web site: www.nhs.co.org Web site: www.
eu/pol/food/index_en.uk/ Society of Food Hygiene and Technology (UK) http://www.int/foodsafety/en/ European Commission: Activities of the European Union food safety http://europa.CONTACTS US FDA .cfsan.cdc.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.FoodSafety.ift.org Web site: http://www.org/ Royal Institute of Public Health (UK) http://www.fda.gov/ World Health Organization (WHO): Food safety programmes and projects http://www.htm Centre for Disease Control and Prevention (CDC) (US) http://www.ift.fpa-food.sofht.gov/foodsafety/ Food Safety Authority of Ireland http://www.riph.org/ The Association of Food.co.who. Beverage and Consumer Products Companies http://www.org.org/ Institute of Food Technologists http://www.gov/ Institute of Food Technologists 525 W.uk/ 245 .fsai.csiro. Van Buren Suite 1000 Chicago IL 60607 United States of America Tel: + 1 3127828424 or + 1 8004383663 Fax: + 1 3127828348 Email: email@example.com/cms/ Useful Web Sites Gateway to Government food safety information (US) http://www.ie/ Food Science Australia http://www.foodscience.au/ International Association for Food Protection http://www.
FISH AND SEAFOOD 246 .
as spoilage organisms in chilled recipe meals 43 growth and survival in chilled and frozen raw fish 17 pathogen profile 225-6 pathogenic microorganisms in molluscan shellfish 59-60. as autochthonous bacterial flora of fish 3 in fish sauce 127 Bacillus cereus. growth and survival in chilled and frozen raw fish 19 of concern in preserved fish 115 Aspergillus spp. in crustacean shellfish 87 Alteromonas spp. as TTX producers 145 Amnesic shellfish poisoning (ASP) toxins 146-8 caused by domoic acid in crustacean shellfish 87 Anchovies. as autochthonous bacterial flora of fish 3 Aeromonas spp. in crustacean shellfish 86-7 ATP. 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 .INDEX Achromobacter spp. as autochthonous bacterial flora of fish 3 as spoilage organisms in oysters 65 as spoilage organisms in temperate crustacean shellfish 80 as spoilage organisms in tropical crustacean shellfish 81 in molluscan shellfish 60 Aerobic plate counts. preserved by dry/wet salting 99 Anisakis spp. planktonic source of spirolides in shellfish 163 Allergens. growth and survival in crustacean shellfish 86 Air drying 101 Alcaligenes. 102 Bacillus spp. growth and survival in crustacean shellfish 86 Aeromonas sobria. causing spoilage of dried fish 110. source of PSP toxins 142 Alexandrium ostenfeldii. to test microbiological criteria 19 Aeromonadaceae. 69-70 Aeromonas hydrophila. as spoilage organism in oysters 65 as spoilage organism in tropical crustacean shellfish 81 in molluscan shellfish 60 Alexandrium. 111 Astroviruses. breakdown of – as indicator of fish freshness 10-1 Azaspiracids 158-9 detection methods 160 origins and distribution 159 toxicology 159-60 Bacalao 97.
68-9 Capture and handling. preserved by marinating 100 Chemical detection methods. growth and survival in crustacean shellfish 86 pathogenic microorganisms in molluscan shellfish 59-60 Campylobacter jejuni. in crustacean shellfish 86-7 Campylobacter coli. description and uses 56-7 effects of processing on microflora 60-4 hot-smoking 107 Clear smoke treatments 108 Clostridium. 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. as cause of spoilage of MAP fish 15 Calciviruses. fish sauce 123 Basipetospora halophila. 226 Breading process 36-8 Brevetoxins. as autochthonous bacterial flora of fish 3 Clostridium botulinum. 136. 137. associated with consumption of fish/fish products 16. 131 Carbon dioxide. 98-9 Brochothrix thermosphacta. of fish – effects on microflora 5-6 Carbohydrate. as gas in modifiedatmosphere packaging 6 CCP Decision Tree (in HACCP plan) (Fig) 185 Ceviche. in crustacean shellfish 87 Chilled prepared fish products 27-51 Chilled raw fish 1-25 Chilled storage. affected by azaspiracids 159 affected by cyclic imines 161 Botulism. of fresh fish spoilage 11 Chemical pollutants. indigenous – in fish (Table) 4 non-indigenous – in fish (Table) 4 Bakasang. cause of neurotoxin shellfish poisoning 153-5 origins and distribution 1545 toxicology 155 importance in wet salting of fish 96 Brining 93. growth and survival in crustacean shellfish 86 growth and survival in molluscan shellfish 59-60. for PSP toxins 144 Chemical indicators. mould causing spoilage of dried fish 110 Battering process 36-8 Bivalves.FISH AND SEAFOOD components of seafood ready meals 40 Bacterial pathogens. importance in lactic acid fermentation in fish products 130. of recipe fish meals 43 Cholera. smoked and dried fish 113-6 hazard in mild-processed marinated products 101 hazard in sun-dried products 102 non-proteolytic – risk of growth in fish recipe meals 44 248 . caused by Vibrio cholerae O1 17 Ciguatoxins 155-8 detection methods 157-8 origins and distribution 1567 toxicology 157 Clams. 113.
of fish – analysis of 10 Diarrhetic shellfish poisoning (DSP) toxins 148-51 detection methods 150-1 origins and distribution 14950 toxicology 150 Dinophysis spp. adverse condition on poorly preserved fish 110 EC grading scheme. 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. to detect DSP toxins 150-1 Enrobing process 36-8 Enterobacteriaceae. description and uses 59 effects of processing on microflora 60-4 Cold-smoking 104-5. of prepared fish products 44 Escherichia coli. for detection of DA 148 ELISA kit (DSP-Check). cooked – effects of processing on microflora 83 forms for purchase 80 freshness indicators 83 Crayfish. smoked and dried fish 114 Enzymic spoilage. hot-smoking 107 ELISA assay. to determine fresh fish spoilage 11 Corynebacteria. growth and survival in crabmeat 84 growth and survival in cured. as autochthonous bacterial flora of fish 3 Crabs. to determine fresh fish spoilage 11 Eels. smoked and dried fish 114 pathogenic microorganisms in molluscan shellfish 59-60 249 . 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. 149-50. as spoilage organisms in tropical crustacean shellfish 81 Coryneforms. growth and survival in chilled and frozen raw fish 19 of concern in cold-smoked salmon 115 Domoic acid (DA). as contaminants in scampi 36. processing 334 Cockles. 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. 152 Diphyllobothrium. forms for purchase 80 Crustacean shellfish 79-92 Cured fish 93-121 Cyclic imines 160-1 detection methods 162 origins and distribution 161 toxicology 162 Degradation. to detect NSP toxins 155 ELISA kit (Biosense). 106 Contacts 241-5 Conway method.INDEX pathogen profile 226-7 Clostridium perfringens. of fish – different processes 94 Dun. source of azaspiracids 159 source of DSP toxins 148-9.
102 Gymnodimines. of fresh fish spoilage 911 Evisceration. of fish at sea 5 Fatty fish. non-B) viruses. causing pink condition on fish surface 110 in fish sauce 127 Halococcus spp. member of cyclic imine group 161 Gymnodinium catenatum. to measure fish skin resistance 10 Flavobacterium. preserved by marinating 100 preserved by dry/wet salting 99 Histamine fish poisoning. fermented fish product 126 Green cured split cod 97. spoilage 9-11 Freshness grading schemes 9 Frozen prepared fish products 27-51 Frozen raw fish 1-25 Frozen storage. in crustacean shellfish 86-7 in molluscan shellfish 59-60 Herring. processing 35 Fish component. 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. specific EU hygiene rules 196-224 Fresh fish. concerning crustacean shellfish 87-8 European Grading Scheme. of fish – effects on microflora 5-6 Hepatitis (non-A. of chilled and frozen prepared fish products – preparation 33-6 Fish sauces 123-4 Fish sauces and pastes. causing pink condition on fish surface 110 in fish sauce 127 Handling. caused by Photobacterium phosphoreum 15 Hon kare bushi 107 Hot-smoking 105-7 250 . preserved by marinating 101 Gravlaks. definition of terms 176-7 implementation and review of plan 184-6 in fish and seafood product manufacture 175-88 seven stages of system 17784 Halobacterium spp.FISH AND SEAFOOD EU legislation. 130-5 Fish-spoilage bacteria 12 Fishtester. for evaluation of fish quality 9 Eurotium. source of PSP toxins 142 Gyoshoyu. of prepared fish products 44 Gambiertoxins. as source of ciguatoxins 156 Garlic. effects of processing 127-30 Fish sauces and pastes of south-east Asia (Table) 124 Fish/salt/carbohydrate products of Asia (Table) 125. mould causing spoilage of dried fish 110 Evaluation. as source of inulin in lactic acid fermentation in fish products 134 Garos. fish sauce 123 HACCP. fermented fish product 124 General Pathogen Surveillance Programme 84 Gravad fish. in crustacean shellfish 86-7 in molluscan shellfish 59-60 Hepatitis A virus. 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 Lean fish. 151-3 detection methods 153 Liquid smoke 107-8 Listeria spp. 131 Lactic fermented fish products (Table) 126 fish species used in the Philippines (Table) 126 Lactobacillus. 135 I-zushi. smoked and dried fish (Table) 116-7 for shellfish products 70 Microbiological limits. preserved by salt-boiling 100 Karenia brevis. 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. outbreak caused by gravid 115 Lobster. enabled by carbohydrate source in fish products 130. definition 1 Legislation.INDEX Hygiene regulations for foodstuffs production and distribution (EU) 190-6 Ice storage. as source of botulism 136 International legislation. cooked – effects of processing on microflora 83 forms for purchase 80 freshness indicators 83 Mackerel. initial – of chilled and frozen prepared fish products 31-2 of chilled and frozen raw fish 3-5 251 . food hygiene 189-224 Lingulodinium polyedrum. for chilled and frozen prepared fish products 47-9 for chilled and frozen raw fish 19-20 for crustacean shellfish 87-8 for cured. in fish fillet 1 Lipid oxidation. for fresh and frozen fish (Table) 20 Micrococcus spp. causing spoilage in vacuum-packed dried salmon 111 Lactic acid fermentation. as source of yessotoxins 152 Lipid content. producer of brevetoxins 154 Katsuobushi 107 Kench curing 97-8 Klipfish 97 Lactic acid bacteria. for microbial inactivation in fish products 127. preserved by salt-boiling 100 Ionising radiation. hot-smoking 107 preserved by dry/wet salting 99 Marinating 94. preserved by marinating 101 Microbiological criteria. concerning crustacean shellfis 88 Jellied eels. smoked and dried fish 114-5 hazard in cold-smoking 105 hazard in mild-processed marinated products 101 Listeriosis. growth and survival in chilled and frozen prepared fish products 46 growth and survival in cured. leading to oxidative rancidity 11 Lipophilic shellfish poisoning toxins 149. 100-1 Masmeen 107 Matjes. as hazard in regard to reef-caught fish 28 growth and survival in crustacean shellfish 84 pathogen profile 229-30 Listeria monocytogenes. to reduce rate of fish spoilage 12 Ikan kaju.
as autochthonous bacterial flora of fish 3 as spoilage organism in temperate crustacean shellfish 80 in molluscan shellfish 60 Mouse bioassay.FISH AND SEAFOOD of crustacean shellfish 80-1 of cured. 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. for detection of azaspiracids 160 for detection of ciguatoxins 157 for detection of DA 148 for detection of DSP toxins 150. in crustacean shellfish 867 Norwalk virus. caused by lipid oxidation 11 Oysters. as TTX producer 145 Norovirus. contamination leading to illness 68 description and uses 57 effects of processing on microflora 60-4 hot-smoking 107 spoilage organisms 65 P. reduction in smoking process 108 Norcardiopsis dassonvillei. of crustacean shellfish – effects of processing 81-3 of cured. contaminated by azaspiracids 158 description and uses 58 effects of processing on microflora 60-4 hot-smoking 107 Nam-budu. growth and survival in chilled and frozen raw fish 17 Paralytic shellfish poisoning (PSP) toxins 142-4 detection methods 144 toxicology 143 252 . smoked and dried fish 95 of fermented fish 126-7 of molluscan shellfish 59-60 Microflora. 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. pathogenic microorganisms in molluscan shellfish 59-60 Opisthorchis viverrini. 153 for detection of LST toxins 151 for detection of NSP toxins 155 for detection of pinnatoxins and pteriatoxins 165 for detection of PSP toxins 144 for detection of spirolides 164 for detection of tetrotoxin 145 Mussels. shigelloides.
145-6 definitions 28-30 non-seafood ingredients 32 Processing. in fish fillet 1 Proteus. in molluscan shellfish 60 Pseudomonas spp. causing spoilage of dried fish 110-1 Photobacterium phosphoreum. reduction in smoking process 103. as component for seafood ready meals 40 Prawns. 163. fermented fish product 124 Plankton. effects of processing on microflora 81-2 freshness indicators 83 Pre-column derivatisation.INDEX Parasites. preserved by salt-boiling 100 Pink. 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. mould causing spoilage of dried fish 110 Potato. as source of toxins 159. 197 Plesiomonas. growth and survival in chilled and frozen raw fish 19 growth and survival in crustacean shellfish 86 growth and survival in cured. pathogen profile 230 in molluscan shellfish 5960. 225-39 Pathogens. contact details 241-2 Prorocentrum spp. for detection of PSP toxins 144. as source of yessotoxins 152 Protoperidinium cassipes. planktonic source of azaspiracids 159 Pseudomonadaceae. 152-3 Penicillium spp. smoked and dried fish 113-6 growth and survival in fermented fish 136-7 growth and survival in molluscan shellfish 66-70 Pectenotoxins 151. smoked and dried fish 115 Pasta. as component for seafood ready meals 40 Pathogen profiles. detection methods 165 origins and distribution 165 toxicology 165 Pissala. adverse condition on poorly preserved fish 110 Pinnatoxins member of cyclic imine group 164. spoilage organism in oysters 65 Protoceratium reticulatum. as autochthonous bacterial flora of fish 3 as spoilage organism in chilled products 43 as spoilage organism in crustacean shellfish 80. as source of DSP toxins 148-9 Protein content. as cause of histamine fish poisoning 15 as spoilage organism in chilled products 43 as spoilage organism in fresh fish 14-5 Pickling 94. 161. 69-70 Polycyclic aromatic hydrocarbons. 108 Polypaecilum pisce. effects on microflora of fish 5-9 Professional bodies. causing spoilage in vacuum-packed dried fish 111 Pseudomonads. 100-1 Pindang. 81 as spoilage organism in fresh fish 13 as spoilage organism in oysters 65 as TTX producers 145 253 .
antibody-based kit to detect STX 144 RT Freshmeter. resulting from poorly salted cod 110 Pyrodinium. in molluscan shellfish 60 Rice. fermented fish product 126 as source of botulism 136 Rakørret. as source of tetrotoxin 144-5 Putty fish. description and uses 58-9 Scampi. flow diagram (Fig. 83 254 . flow diagram (Fig. 135 Rakefisk.FISH AND SEAFOOD inhibition of growth by carbon dioxide 6 Pseudonitzschia multiseries. hazard in salting of fish 97 Salt concentration. processing and effects on microflora 60-1 Shewanella. preserved in saturated brine 99 Sauce. detected in cod 115 Pteriatoxins. to measure fish skin resistance 10 Salmon. producers of brevetoxins 154 Raw fish. assembly and packaging. fermented fish product 126 Raphidophytes. detection methods 165 member of cyclic imine group 164 origins and distribution 165 toxicology 165 Puffer fish. smoked and dried fish 114 pathogen profile 230-1 pathogenic microorganisms in molluscan shellfish 59-60 Salt burn. growth and survival in chilled and frozen raw fish 19 Pseudoterranova dicipiens. 100 Salting 97-9 definitions 93-4 Sardines. processing 34. smoking and marinating of fish 96 Salt-boiling 94. as autochthonous bacterial flora of fish 3 inhibition of growth by carbon dioxide 6 Shewanella putrefaciens. as spoilage organism in chilled products 43 as spoilage organism in fresh fish 13-4 as spoilage organism in temperate crustacean shellfish 80. implicated in amnesic shellfish poisoning 146-7 Pseudoterranova.) 41 ingredients preparation. as hazard in regard to reef-caught fish 28 growth and survival in crustacean shellfish 84 growth and survival in cured. source of PSP toxins 142 Quality Index Method.) 41 Recipe fish products 38-40 Rhodotorula rubra. importance in lactic acid fermentation in fish products 130 importance in salting. vacuum packing 105 Salmonella spp. for microbial inactivation in fish products 127. 36 Seafood. 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. for evaluation of fish quality 9 Radiation. processing 35 Shellstock. for fish ready meals – manufacture 40-2 Saxitoxin (STX) 142-3 Scallops. initial microflora 31 Seafood ready meals 39-40 Seafood recipe meal centres 38-9 Shaped fish products.
fish and shellfish 141-74 Trade associations. to measure fish skin resistance 10 Total Volatile Base-Nitrogen. 13. of cooked fish 9-10 Tetrotoxin 144-6 detection methods 145-6 toxicology 145 TMAO. as indicator of fresh fish spoilage 11 Toxins. fermented fish product 126 Taste panel evaluation. in molluscan shellfish 60 Tuna. in poorly salted cod 110 Smoked fish 93-121 Smoking 103-8 different processes 94-5 Snow Mountain virus. 12-5 of recipe fish meals 43-4 of shellfish 65 Staphylococcus. definitions of forms for purchase 79-80 effects of processing on microflora 81-2 freshness indicators 83 Shucked shellfish. role in fish degradation 10. pathogenic microorganisms in molluscan shellfish 59-60 Specific spoilage organisms (SSOs) 12-5 Spirolides. smoked and dried fish 114 pathogen profile 232-3 pathogenic microorganisms in molluscan shellfish 59-60 potential for growth in pasta component of seafood ready meals 40 Stockfish. as contaminant in scampi 36 as contaminant in enrobing process 37 as hazard in dry salting of fish 89 growth and survival in chilled and frozen prepared fish products 46-7 growth and survival in crustacean shellfish 84 growth and survival in cured. contact details 241-2 Trematode infections foodborne 137 Trichosporon. 83 of fermented fish 135-6 of fish subjected to different preservation methods 108-10 of fresh fish 9-11. 101 Storage. of dried fish 111 of fish 6-7 of fish – (US) NACMCF recommendations 7-8 of fish – Codex Committee 255 . for evaluation of fish quality 9 Torrymeter. of crustacean shellfish 80. grading of 31 hot-smoking 107 US legislation. concerning crustacean shellfish 89 Vacuum packaging. 14 Torry Scale. as measure of fish degradation 10 Sliming. of fish after capture 5 Surstrømming. growth and survival in chilled and frozen prepared fish products 46 growth and survival in cured. processing and effects on microflora 63-4 Skin resistance. smoked and dried fish 114 pathogenic microorganisms in molluscan shellfish 59-60 Shottsuru.INDEX Shigella spp. fish sauce 123 Shrimps. detection methods 164 member of cyclic imine group 162-4 origins and distribution 163 toxicology 163-5 Spoilage.
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. 68 Vibrio cholerae non-01. growth and survival in molluscan shellfish 67-8 Vibrio parahaemolyticus. as autochthonous bacterial flora of fish 3 as spoilage organisms in chilled products 43 as spoilage organisms in vacuum-packed dried fish 111 Viruses. 81 as TTX producers 145 pathogenic microorganisms in molluscan shellfish 59-60 risk in cold-smoking 105 Vibrio cholerae. as spoilage organisms in chilled recipe meals 43 Yessotoxins 151. 62 Yersinia spp. growth and survival in chilled and frozen raw fish 17 Vibrio cholerae 01 and 0139. 152-3 256 . processing for seafood recipe meal centres 39 Vibrio spp. growth and survival in chilled and frozen raw fish 16-7 growth and survival in crustacean shellfish 86 growth and survival in molluscan shellfish 67 pathogen profile 235-6 Vibrionaceae.FISH AND SEAFOOD guidance 7 of fresh chilled products 42 Vegetables. causing dun condition on dried fish 110 Wet salting 98-9 Wet storage. of shellstock 61. growth and survival in chilled and frozen prepared fish products 47 growth and survival in crustacean shellfish 86-7 Wallemia sebi. growth and survival in crustacean shellfish 85 pathogen profile 233-4 Vibrio cholerae 01. as hazard in regard to reef-caught fish 28 as spoilage organism in crustacean shellfish 80.
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