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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
Goldwyn. The books in the series are designed as easy-to-use guides to the microorganisms found in foods. and a detailed list of contacts for various food authorities.FOREWORD The Microbiology Handbook series includes Dairy Products. Training Manager. I dedicate this book to my husband . for his scientific input and Victoria Emerton. and Ann Pernet for the work put into typesetting and indexing. which has been the chief factor in ensuring the completion of this book. potential hazards associated with the consumption of a range of products. Each book provides a brief overview of the processing factors that determine the nature and extent of microbial growth and survival in the product. and growth characteristics for key pathogens associated with the product. thank you for all the support. published by Leatherhead Food International and RSC Publishing. both in the UK and elsewhere. All handbooks also contain a review of the related legislation in Europe and UK. Acknowledgements All the authors involved in writing the first edition of this book are gratefully acknowledged. I also acknowledge and thank Jackie Apps. The books are intended as a source of information for microbiologists and food scientists working in the food industry and responsible for food safety. I thank the contributing authors for their collaboration. Fish and Seafood. Rhea Fernandes Leatherhead Food International v . Alison Turner. guides to HACCP. Technical Team Leader for her valuable editorial contribution. My gratitude is extended to Dr Peter Wareing. and Meat Products.
7 Published Microbiological Criteria 2.8 References MOLLUSCAN SHELLFISH 3.6 Pathogens: Growth and Survival 1.2 Initial Microflora 1.4 Processing and its Effects on the Microflora 2.3 Processing and its Effects on the Microflora 1.4 Spoilage 1.CONTENTS CONTRIBUTORS FOREWORD INTRODUCTION 1.3 Processing and its Effects on the Microflora 3.6 Pathogens: Growth and Survival 2.3 Processing and its Effects on the Microflora 4.4 Spoilage 4.4 Spoilage 3.2 Initial Microflora 3.5 Spoilage 2.7 References 4.1 Definitions 1. vii .2 Definitions 2.8 Further Reading CRUSTACEAN SHELLFISH 4.5 Pathogens: Growth and Survival 3.2 Initial Microflora 4. CHILLED AND FROZEN RAW FISH 1.7 References 3.1 Introduction 2.9 Further Reading CHILLED AND FROZEN PREPARED FISH PRODUCTS 2.7 Published Microbiological Criteria 1.8 Further Reading iii v xi 1 1 3 5 9 12 15 19 20 24 27 27 28 31 32 43 45 47 49 53 53 59 60 65 66 70 71 77 79 79 80 81 83 84 87 89 92 2.1 Definitions 4.3 Initial Microflora 2.6 Published Microbiological Criteria 3.1 Definitions 3.5 Pathogens: Growth and Survival 4.6 Published Microbiological Criteria 4. 3.8 References 1. 4.5 Factors Affecting Fresh Fish Spoilage 1.
7.2 Definitions 8.I.3 Tetrodotoxin 7.1 Introduction 7. S.2 Legislative Structure 190 9.3 Processing and its Effects on the Microflora 5.1 Definitions 5.12 References HACCP IN FISH AND SEAFOOD PRODUCT MANUFACTURE 8.11 Conclusion 7.4 Spoilage 5.7 Food Hygiene (England) Regulations 2006.6 Lipophilic Shellfish Toxins (LST) 7.8 Further Reading FERMENTED FISH 6. 9.5 References 93 93 95 96 108 113 116 117 120 123 123 126 127 135 136 137 140 141 141 142 144 146 148 151 153 155 158 160 165 166 175 175 176 177 184 186 6.5.4 Regulation (EC) No.6 Published Microbiological Criteria 5.1 Introduction 189 9.8 Ciguatoxins 7.7 Further Reading FISH AND SHELLFISH TOXINS 7. 852/2004 on the General Hygiene of Foodstuffs 191 9.1 Definitions 6.10 Further Reading 223 viii .8 Guidance 222 9. 14 (Hygiene requirements specific to the UK) 221 9.9 Azaspiracids 7.3 Stages of a HACCP Study 8.1 Introduction 8.4 Amnesic Shellfish Poisoning (ASP) Toxins 7.6 References 6. 2006 No.5 Diarrhetic Shellfish Poisoning (DSP) Toxins 7.2 Paralytic Shellfish Poisoning (PSP) Toxins 7.5 Pathogens: Growth and Survival 6.4 Spoilage 6.10 Cyclic Imines 7.3 Regulation (EC) No.2 Initial Microflora 5.2 Initial Microflora 6.7 Neurotoxin Shellfish Poisoning (NSP) Toxins 7.6 Regulation (EC) No.5 Regulation (EC) No. SMOKED AND DRIED FISH 5. 854/2004 of the European Parliament and of the Council Laying Down Specific Rules for the Organisation of Official Controls on Products of Animal Origin Intended for Human Consumption 212 9. 2073/2005 on Microbiological Criteria for Foodstuffs 214 9. EU FOOD HYGIENE LEGISLATION 189 9. 8.7 References 5. 853/2004 Laying Down Specific Hygiene Rules for Food of Animal Origin 196 9.9 Other Relevant Legislation 223 9. CURED.4 Implementation and Review of the HACCP Plan 8.5 Pathogens: Growth and Survival 5.3 Processing and its Effects on the Microflora 6.
7 Staphylococcus aureus 10. 10.2 Clostridium botulinum 10.9 Vibrio parahaemolyticus 10.8 Vibrio cholerae 10.10 PATHOGEN PROFILES 10.3 Clostridium perfringens 10.1 Aeromonas spp.4 Listeria spp.6 Salmonella spp.11 References 225 225 226 227 229 230 230 232 233 234 235 236 241 247 CONTACTS INDEX ix . 10. 10.10 Vibrio vulnificus 10.5 Plesiomonas 10.
Milk. The Microbiology Handbook. Because of their health advantages over red meats.. Oehlenschlager J. rivers and lakes whose water can range from pristine to contaminated.. Hoboken. 2007. Jacobsen C. Nollet L. in Handbook of Food Products Manufacturing: Health.. Meat.INTRODUCTION Fish and seafood are a main source of animal protein in the diet.. Often contamination is from human and animal sources. Poultry. Wiley. Seafood processing: basic sanitation practices. Clark S.. Geographical region.H. 94767. Further Reading General Fish/Seafood Microbiology Cross N. Saebo A. and fish and shellfish toxins. Key changes in this edition are the recent regulatory changes pertaining to food hygiene and microbiological criteria for foodstuffs. Cross N.A. thus. Wageningen. molluscan shellfish. and. Safety and Processing of Wild and Farmed Fish. 2006. Hui Y. Catches can be gathered from seas. the consumption of fish and seafood has increased. crustacean shellfish. The second edition of this handbook is a review of the entire book for currency of information. Eds. Chandan R.Fish and Seafood consists of the microbiology of seven different product categories: chilled and frozen raw fish. season.B. cured. Seafood Research from Fish to Dish: Quality. fish and seafood can be involved in the transmission of pathogenic microorganisms and toxins..M. smoked and dried fish.. Wageningen Academic Publishers. Bekaert K.. chilled and frozen prepared fish. Seafood and Vegetables. xi . whether they are pelagic (surface to mid-water) or demersal (bottom) feeders will influence the numbers and types of microorganisms present on freshly caught seafood. Luten J. for fish. fermented fish.
Rawles D.. Modified-atmosphere packaging of fish and fish products. Stanfield P. Minimal processing in practice: seafood. Salmonella spp. Ed. Gudmundsson M. 1998. International Commission on Microbiological Specifications for Foods. Hui Y.A.M. Elsevier.S.H. Eds. CRC Critical Reviews in Food Science and Nutrition. Hafsteinsson H. in Minimal Processing Technologies in the Food Industry. in Food Processing: Principles and Applications. Ed. 121-37.R. basic sanitation practices. 2003. Eds. Ed. Simpson B. 1997. London. Flick G. Huss H.P. Amsterdam.International Commission on Microbiological Specifications for Foods. 318-23.M. Vo1ume 39. 1997.L.Y. 8 (2). Blackwell Publishing.E. Gorham J. 1996.H. Taylor S. Gram L.. 1996. 2004. Proceedings of the International Seafood Conference. Woodhead Publishing Ltd.N. Academic Press.B. Huss H. Elsevier.A.H. 167-77. 1996.. in Food Plant Sanitation. Davies A. McCarthy S. Smith J.S. Ventresca P. 459-72.. Oxford. Marcel Dekker. Food Control. 245-66 Bremner H. November 1995. Gram L.R. xii ..A. 174-249. Johnson J. New York. 200-23. Blackie. Cambridge. Noordwijkerhout. 2002. Eds.. Microbiological spoilage of fish and fish products. The fish industry. in Fish Processing Technology. London. Incidence and survival of Listeria monocytogenes in ready-to-eat seafood products. and Clostridium botulinum in smoked fish and shellfish.. in Microorganisms in Foods 6: Microbial Ecology of Food Commodities. Nip W-K. Luten J. Noordwijkerhout. 61 (3).H. 1997.. 372-6. November 1995. Tong P. Control of indigenous pathogenic bacteria in seafood. Heinitz M. Ed. Proceedings of the International Seafood Conference.L. 91-8.. Ed. The incidence of Listeria spp. International Journal of Food Microbiology. Stanfield P. Journal of Food Protection... 2002..Y. Hui Y. Spoilage and shelf-life extension of fresh fish and shellfish.. Seafood: processing. in Kosher Food Production. Journal of Food Protection.K. Martin R.. in Seafood from Producer to Consumer: Integrated Approach to Quality. 60 (4). 2004. Ohlsson T. 1997.L. 329-65. Dalgaard P. 431-43. 413-30. 2005. 543-61. Kluwer Academic / Plenum Publishers. 33 (1). Luten J. Woodhead Publishing Ltd. Amsterdam. Cambridge. Ed. Huss H. Oxford.. Blech Z. Hall G. Bengtsson N. Biogenic amines in fish and shellfish. 87121. 36 (1-2).. Ashie I. Dalgaard P.D. Bruinsma B. Smith J.. Seafood processing: basic sanitation practices.H.. 1997. Microbiology of fish and fish products. New York. in Seafood from Producer to Consumer: Integrated Approach to Quality.J. Predictive microbiological modelling and seafood quality. Blech Z.B. Blackwell Publishing. Fish and fish products (Microorganisms in fish products). Safety and Quality Issues in Fish Processing. in Advances in Food and Nutrition Research.
Institut fur Veterinarmedizin. Bacterial pathogens transmitted by seafood. Hobbs G.R. PHLS Microbiology Digest. Microbial growth and its control in meat.. Bacterial spoilage of seafood. A new look at microbiological standards. Elsevier. 1995. Elsevier. 1-17. 26 (3).M. 1992. in Microorganisms in Foods. 1995. International Journal of Food Microbiology. Elsevier. Jakobsen M.H. Microbiological indicators and seafood processing. Pearson A. Huss H. Qualitative and quantitative characterization of spoilage bacteria from packed fish. in Quality Assurance in the Fish Industry: Proceedings of an International Conference. Eds. 44 (12). Copenhagen. 269-82. Food Science and Technology Today. 1990. Copenhagen. 319-33. Gram L. Bernoth E-M. Ed. 1994. Food Technology. Ward D. 1992.. Ed. London. Martin A.. Sofos J. Dalgaard P. 118-20. London. Gibson D. Olley J. Amsterdam. August 1991. 1989. Fish spoilage and safety. in Quality Assurance in the Fish Industry: Proceedings of an International Conference. in Quality Attributes and Their Measurement in Meat. 11 (2). International Journal of Food Microbiology. Ward D. Activities Report of the R&D Associates. Amsterdam. 90-106. 1991.N. 5 (3). in Fisheries Processing: Biotechnological Applications. 1994. Microbial hazards of seafood consumption. Poultry and Fish Products.. 1994.R.R. in Quality Assurance in Seafood Processing: A Practical Guide. Gram L. 265-79.. 305-17. Evaluation of the bacteriological quality of seafood.. McMeekin T. Sterling Publications Ltd. Seafood toxins of microbiological origin. Berlin. Application of predictive microbiology to assure the quality and safety of fish and fish products. 459-78. Twigg B. Eds.. 25-39. Ross T.D. Amsterdam. 1992. Fish: microbiological spoilage and safety. Liston J. in Food and Beverage Technology International: USA. 54-61.D.International Commission on Microbiological Specifications for Foods. Garrett E. Eds. Volume 5: Microbiological Specifications of Food Pathogens. 1996.H.. Dalgaard P. Liston J. guidelines. Liston J.S. Blackie. 16 (1). in Quality Assurance in the Fish Industry: Proceedings of an International Conference.H. 359-403. 2257. Huss H. 166-73. Glasgow. Chapman and Hall. Liston J. 1992. Chapman & Hall. 121-8. London. Eds. Microbiological quality of fishery products. 1990. Gilbert R. 5662. International Journal of Food Microbiology. Ed.J. 1994. Evaluation of the bacteriological quality of seafood. August 1991. Bonnell A. Huss H. 26 (3). 1994. Bonnell A.M.A. Jakobsen M. Liston J. London. Ed. poultry and fish. Ed. in Public Health Aspects of Seafood-borne Zoonotic Diseases. 93-105. xiii . and specifications for seafood products. International Commission on Microbiological Specifications for Foods.A. August 1991. Blackie. Modelling of microbial activity and prediction of shelf-life for packed fish. Dutson T. Copenhagen. 46 (1). The microbiology of seafood products. Jakobsen M.M..
and specifications and inspection of seafood products.a review. Regenstein J. 1982.S.. 46 (1). Lamprecht E.K. Part I . Journal of Food Quality. Ed.spoilage of fish. 1986. Norris J. Ed. 1988. in Microbiology of Frozen Foods. in Developments in Food Microbiology. 42 (3).Pedrosa-Menabrito A. Hobbs G. Chichester. Microbiological hazards associated with fishery products. Wiley. Microbiological standards. 1987.K. in Essays in Agricultural and Food Microbiology. 1985. 103. guidelines. 42 (3). London. Applied Science Publishers.M. 11 (2). xiv . Shelf-life extension of fresh fish . 71-117. 8-16.J. Davies R. Simmonds C..C. Hodgkiss W. CSIRO Food Research Quarterly. London. Hackney C. Ed. 199-226. Elsevier Applied Science Publishers. Seafood-borne bacterial pathogens of marine origin.. The bacteriology of fish handling and processing. 169-208.. Eyles M. Microbiology of fish.R. Microbiology of frozen fish and related products.R. 90-3. 1988. Hobbs G. Garrett E. Volume 1. Robinson R. 117-27. 1988. Dicharry A. 104-9. Food Technology. Food Technology.
while fatty fish have fat cells distributed along their bodies. which can be up to 67%. offering less than 0. Fish fillets are a poor source of carbohydrates. Protein content in fish fillet varies typically from 16 . Fish flesh is dominated by the abundance of white muscle in relatively short segments.21%. most fish contain omega 3-fatty acids and other essential nutrients.1. typically fluctuates between 0. Although fish is broadly similar in composition and structure to meat there are a number of distinctive features. and excludes molluscs and Crustacea. some regulatory agencies such as the US Food and Drug Administration (FDA) will include molluscan shellfish. giving it its characteristically flaky structure. and is mostly interspersed between the muscle fibres. The lipid content. it shows a typical season pattern especially in migratory species such as herring or mackerel. crustaceans. The lipid fraction is the component showing the greatest variation. The connective tissue content of fish is also lower than that found in meat. In addition. 1 .20%. respectively (1). Fish can be divided into fatty and lean fish. and other forms of aquatic animal as part of their ‘fish’ definition. Muscle composition and structure of fish also differ from those found in other meat. lean fish are those fish that store most of their fat in the liver. CHILLED AND FROZEN RAW FISH Associate Prof. and spawning season. fins and a streamline body. Fish are an important part of a healthy diet since they contain high quality protein. Covadonga Arias Department of Fisheries and Allied Aquacultures Auburn University Auburn Alabama 36849 United States of America 1. ‘fish’ also refers to the flesh of such animals used as food. but typically present a low fat percent when compared to other meats. In this chapter. However.2 . This super class of vertebrates includes all the bony and cartilaginous finfish. In addition.5% (1). “fish” will be used for fresh and seawater finfish. typically 3 and 15% of total weight.1 Definitions Fish are classified as any of the cold-blooded aquatic vertebrates of the super class Pisces typically showing gills. migratory patterns. Fish fillet composition can vary significantly within the same species due to feed intake.
DMA is dimethyl amine. but no further control is exercised during storage. at -2 to -4 °C. hence. Fresh fish is raw fish that has not been processed. transportation and sale. Shelf life is defined as the time of storage before microbial spoilage of a fish is evident. renders that fish objectionable to consumers. and flavour based on the maturity. and maintained at or below -2 °C (normally below -12 °C) during storage. nitrogen (N2) and oxygen (O2). Controlled-atmosphere packaging (CAP) refers to packaging in an atmosphere where the composition of the gases is continuously controlled during storage. This technique is primarily used for bulk storage. The proportion of each component is fixed when the mixture is introduced. Evisceration is the removal of the viscera from a fish. juiciness. transportation and sale. Spoilage potential is a measure of the propensity of microorganisms to render fish objectionable to consumers through the production of offensive metabolic byproducts. odour and texture.FISH AND SEAFOOD Chilled fish is fish that has been cooled to. colour. Quality refers to palatability and organoleptic characteristics such as tenderness. transportation and sale. Modified-atmosphere packaging (MAP) refers to packaging systems in which the natural gaseous environment around the product is intentionally replaced by other gases. and texture of the fish. spoilage microflora describes an association of microorganisms that. during storage. Organoleptic refers to qualities such as appearance. colour. usually carbon dioxide (CO2). and maintained at temperatures just below the freezing point. Spoilage describes changes that render fish objectionable to consumers. Raw fish refers to fish that has not been cooked but excludes fish treated with curing salts and/or subjected to fermentation. firmness. Frozen fish is fish that has been cooled to. 2 . but not below 3 °C during storage. frozen or preserved. marbling. Superchilled fish is fish that has been cooled to. through their development on fish. and maintained at or below 7 °C.
1. Fish harvested from clean and cold waters will present lower bacterial numbers than fish from eutrophic and/or warm waters. posing a concern for seafoodborne illnesses (2). Flavobacterium.II) (3). Clostridium.I). Human pathogenic bacteria can be part of the initial microflora of fish. Micrococcus. healthy fish is considered sterile and should not present any bacteria or other microorganisms. the natural fish microflora tends to reflect the microbial communities of the surrounding waters.CHILLED AND FROZEN RAW FISH Vacuum packaging (VP) refers to packaging systems in which the air is evacuated and the package sealed. However. It is difficult to estimate how many microorganisms are typically associated with fish. In general. several culture media containing sodium chloride and more than one incubation temperature should be used. and those that although not autochthonous to the aquatic environment. are present there as result of contamination (anthropomorphic origin or other) or are introduced to the fish during harvest. Moraxella. More accurate. It must be noted that mesophilic bacteria can rapidly overgrow psychrotropic organisms. Gastrointestinal tracts and gills typically yield high bacteria numbers. In fact. It is crucial to mimic the environmental physicochemical parameters when isolating bacteria from fish. potential human pathogens may be present in both scenarios. The autochthonous bacterial flora of fish is dominated by Gram-negative genera including: Acinetobacter. Members of the families Vibrionaceae (Vibrio and Photobacterium) and the Aeromonadaceae (Aeromonas spp. as with other vertebrates. Lactobacillus and coryneforms can also be found in varying proportions (1). Shewanella and Pseudomonas. 3 . For example. These pathogens can be divided into two groups: organisms naturally present on fish (Table 1.) are also common aquatic bacteria. The number and diversity of microbes associated with fish depend on the geographical location. gills and the gastrointestinal tract of fish. standard culture-dependent methods can only recover between 1 to 10% of total bacteria present in any given sample. some species (most Vibrios) require sodium chloride for growth. and typical of the fish flora. processing or storage (Table 1. Gram-positive organisms such as Bacillus. although these are influenced by water quality and feed. whenever possible. molecular-based methods have not yet been used to address this issue.2 Initial Microflora The subsurface flesh of live. On the contrary. the season and the method of harvest. microorganisms colonise the skin. since they heavily depend on the type of sample analysed and the protocol used for isolation.
human origin Minimum Infective Dose Variable depending on the strain (>102 cells/g) Staphylococcus aureus 105 .35 °C 8 . birds.106 cells/g (exception: V. and mud Ubiquitous. Plesiomonas shigelloides 3 .37 °C High for most species. 105 .103 High (107 .19 µg/kg bodyweight from < 102 .0.1 . Toxin levels 0.14 .106 cells/g.FISH AND SEAFOOD TABLE 1. 4). sewage. vulnificus) 5 .26 °C 10 .>106 Salmonella spp. In addition. and risk associated with the consumption of seafood is low (2.II Non-indigenous bacterial pathogens frequently present in fish (3) Organism Listeria monocytogenes* Primary habitat Soil. Vibrio cholerae Vibrio parahaemolyticus Vibrio vulnificus other vibrios Aeromonas spp. during storage indigenous spoilage bacteria tend to outgrow potential pathogenic bacteria.1 µg toxin lethal dose Clostridium botulinum non-proteolytic type E Pathogenic Vibrio spp. estuarine environments. Intestinal track of terrestrial vertebrates Human origin Fecal contamination Ubiquitous in environment Shigella spp. However.I Indigenous bacterial pathogens typically present in fish (3) Organism Temperature range Estimated Minimum Infective Dose 00.109 cells/g) * Some sources considered L. numbers of pathogenic bacteria in raw fish tend to be low.37 °C Unknown Unknown TABLE 1. Escherichia coli Yersinia enterocolitica 101 . stream water.102 101 . 4 . 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.
After capture. in which the bacterial load 5 . and their re-use will increase the crosscontamination of other fish with such microorganisms. in which the health of each animal can be assessed prior to slaughter. Of particular concern is the use of wooden or soiled plastic containers for storage and unloading at the quayside. in recent decades. handling and processing Wild finfish are usually caught by net. bruising and bursting of the gut will increase the number of sites open for bacterial attack and spread. The length of time that set nets have been in the water or the time trawlers’ nets are towed. chilled brine or refrigerated seawater at -2 °C. finfish will almost invariably come into contact with nets. aquaculture practices have been expanding worldwide. These contacts will not only increase the bacterial cross-contamination between fish batches but will introduce microorganisms from other sources such as humans.1 Processing and its Effects on the Microflora Capture. and spoilage. decks. and the killing process is designed to minimise stress. the cuttings provide areas of exposed flesh that are open to microbial attack.CHILLED AND FROZEN RAW FISH Shelf life depends on the initial microflora on the fish. care should be taken in removing all the gut contents and washing the carcass thoroughly prior to refrigerating. offering better control of fish health prior to. or traps. If evisceration is carried out at sea. and especially when fish is stored on board for longer periods.3 1. human hands and clothing. mackerel or herring. boxes and/or baskets. with fish such as tuna and cod being more commonly eviscerated than sardines. Used refrigerated brines can be contaminated with high numbers of psychrotrophic spoilage bacteria. cortisone levels increase during prolonged stress and can alter the fillet quality. icing or freezing. During capture and storage. Physical damage such as loss of scales. has an effect on the amount of stress and physical damage that the fish will suffer during capture. Inadequate circulation of chilled brines may result in localised anaerobic growth of some microorganisms. digestion and off-flavours in uneviscerated fish.3. However. Fish may be eviscerated prior to storage at sea . The decision to eviscerate the catch at sea will depend greatly on the size of the fish and the duration of storage at sea. and conditions of storage. ropes. potential contaminants added during handling and processing. with very little control over the condition of the fish at the time of death or the duration of the killing process. however. This contrasts greatly with the meat industry. the fish may be stored in the vessel for periods ranging from just a few hours to several weeks in melting ice. freezing facilities (-18 °C) may be used to prevent the catch from deteriorating.a practice that may have both advantages and disadvantages. 1. hook and line. Increasingly. The action of intestinal enzymes and activity of the gut bacteria on the flesh around the belly cavity may produce discolouration. with the production of off-odours. and during harvest. In addition. In eviscerated fish. birds and soil.
however. What is more.3. but isolate the food in an environment different from the bulk atmosphere. oxygen etc. Packaging changes the intrinsic and extrinsic parameters affecting a product. not part of the normal flora of the meat animals (8). Both carbon dioxide and low pH inhibit the growth of the typical spoilage bacteria such as Pseudomonas and Shewanella. by actively altering the composition of the 6 . By vacuum packing foods. In the absence of oxygen. allowing more growth of spoilage organisms.FISH AND SEAFOOD can be substantial. careful and sanitary handling during processing is required to reduce the risk of contamination with potential human pathogens. ferric ion (Fe3+)). nitrogen. These containers are also used for displaying the fish during auction at the quayside. potential spoilage bacteria are among the psychrophilic and psychrotrophic flora present on temperate-water fish before death (9). they are usually process contaminants. if there is some oxygen demand from the product. 7). The atmospheric conditions surrounding a product may be passively or actively altered. and of the quality of the water used for ice and to wash fish. in time. but if applied properly should extend the life of the product. and specific spoilage organisms may use other terminal electron acceptors naturally present in the fish (trimethylamine-N-oxide (TMAO). Carbon dioxide is the most important gas in MAP of fish because of its bacteriostatic and fungistatic properties. the natural atmosphere is replaced with a controlled gas mixture (carbon dioxide. Anaerobic atmospheres have less effect on fresh fish shelf life. often in the absence of adequate refrigeration. However. In MAP. fish have a higher post mortem pH. As with all foods. Good Manufacturing Practice (GMP) and control of the sanitary conditions of the transport and processing environments are essential to limit additional risk of disease caused by fish consumption (6. Brochothrix thermosphacta and psychrotrophic lactic acid bacteria (LAB) can produce spoilage characteristics.). will result in fully anoxic conditions.2 Modified-atmosphere packaging A natural atmosphere rich in oxygen (21%) is responsible for oxidative processes and for all aerobic respiratory life. use of effective detergents and disinfectants. MAP extends the shelf life of most fishery products by inhibiting bacterial growth and autoxidation. for example. partial fermentation of sugars occur leading to lower pH. Low oxygen levels have been shown to substantially prolong the freshness and quality life of refrigerated seafood products. a reduction in the oxygen tension is achieved which. and minimising handling will all reduce microbial cross-contamination. 1. Physical barriers not only protect from physical damage. Monitoring of the seawater for algal growth in order to limit the risk of algal toxin ingestion. These changes can be deleterious. Bacterial composition under MAP shifts from mostly Gram-negative to predominantly Gram-positive (lactic) bacteria. cleaning of the work environment. and to limit the loss of quality (5). from water activity (aw) through to physical damage.
Growth and toxin production have been detected in artificially contaminated packs of whole trout after 1 week’s incubation at 10 °C (13). In the United States. which provides guidance relating to vacuum or modified-atmosphere packaging for specific fish products (14). after additive application. phosphates. The (International) Codex Committee has published a Code of Practice for Fish and Fishery Products (CAC/RCP 52-2003). an inert and odourless gas. and the problems of muscle structure damage by the modified-atmosphere (11). MAP has been applied to fresh meat and fish with a resulting commercially viable extension in shelf life (10).3 °C and produce toxins. a modified-atmosphere may contain any gas necessary for the desired effect. nitrous oxide and carbon monoxide have also been suggested as possible replacement gases in trace amounts for MAP/CAP. sorbates and chelating agents such as ethylenediaminetetraacetic acid (EDTA) (11). Of particular concern are the psychrotrophic type E and non-proteolytic type B and F strains. The products. including salt. but residual oxygen and other electron acceptors may be sufficient to allow oxidative spoilage of fish (12). the inhibition of common 'spoilage' bacteria (Pseudomonas. High concentrations of carbon dioxide have the most pronounced microbial effect. as they are able to grow at temperatures as low as 3. Replacement of oxygen with nitrogen.CHILLED AND FROZEN RAW FISH surrounding gas. the higher pH and reduction potential (Eh) of the fish muscle. Many additives have been tried in conjunction with changed atmosphere. which are able to grow rapidly at low temperature. may not be considered ‘fresh’ fish. and the MAP of fish has more challenges to overcome as a result of: a comparatively large initial load of bacteria present. Gases used in MAP of fish most commonly include carbon dioxide and nitrogen. the pathogens that may be able to grow before spoilage occurs. does inhibit some aerobic bacteria and reduce the rate of oxidative rancidity. and increase in-pack drip (12). without overt signs of spoilage. but can dissolve into fish liquids and deform packages. Sulphur dioxide. This has been driven partly by increased consumer demand for fresh and chilled convenience foods containing fewer chemical preservatives. discolour pigmented fish (11). Modified-atmosphere preservation of fish was first reported in the 1930s. The single most important concern with respect to the use of MAP is the potential for outgrowth and toxin production by C. The principal effect of raised carbon dioxide-MAP is an extension of the 'lag' phase of the growth of the bacteria on the fish. VP is one of the oldest forms of altering the interior gaseous environment of a pack. The microflora of meat is not the same as that of whole. Micrococcus and Moraxella). botulinum. and the promotion of a predominantly Gram-positive. gutted or filleted fish. the National Advisory Committee on Microbiological Criteria for Foods (NACMCF) has published a number of recommendations on 7 . although less information on their effectiveness is available. but only in recent years has it seen a marked expansion in use and market share. Flavobacterium. slower-growing flora (11).
use of high-quality raw fish. oxidative rancidity is a significant source of spoilage and hence the complete removal of oxygen and the use of oxygen-impermeable films are preferred. 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. adequate shelf life and cooking requirements. It highlights temperature control as the primary preventive measure against the possible hazard of toxin production by C. 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). at all times. The growth of these bacteria is reduced by high carbon dioxide concentrations but. and the use of a HACCP plan. owing to the dissolution of cellular structures.FISH AND SEAFOOD the safety of MAP and VP refrigerated raw fishery products (15). grouping of fish types is possible to estimate the best carbon dioxide level to be used in MAP. It was also noted that organoleptic spoilage and rejection by the consumer should occur before the possibility of toxin production.g. 5. from where drip loss and gaping of the muscle meant that organoleptically classified rejection occurred sooner than with no carbon dioxide addition (12). greater advantages are seen. In the United Kingdom. from both a microbiological and a chemical standpoint. as already stated. this problem occurs to a lesser extent with meat. but is recognised as a drawback of carbon dioxide application. Only the highest-quality fish. Freshwater fish have different intrinsic bacterial flora and also do not contain such high concentrations of amines (e. should be used. The replacement gas should contain at least 40% carbon dioxide.g. For fatty fish. The only processing dip to be used is 5% potassium sorbate and 10% sodium tripolyphosphate. may result in unacceptable colour and textural change and hence little shelf life extension (11). 4. adequate product labelling in respect of storage temperature. the Sea Fish Industry Authority has published guidelines for the handling of fish packaged in a controlled-atmosphere (16). For other less fatty marine fish. Although all fish species need to be treated separately. Subsequent storage must be at or below 3 °C. 2. 3.3 °C at all points from packaging onwards. The 1985 British Guidelines on MAP/CAP fish from the National Fisheries Institute state that: 1. rejection due to alteration in fish texture with greater carbon dioxide makes this unsuitable (13). leading to the recommendation that the sale of MAP/VP raw fishery products be allowed only when certain conditions are met. Sensitive and accurate time-temperature indicators must be used to ensure that the product has not been temperature-abused. trimethylamine and sulfides). botulinum. These conditions include storage of the product at ≤ 3. however. The problem of water loss in MAP fish is common. however. TMAO) that can be reduced to produce 8 . The addition of high levels of carbon dioxide to these fish.
III. rejection of whole fresh fish for human consumption may be made without chemical or microbiological evaluation. with no smell.4 1. the skin turns from iridescent to dull and bleached with bacterial slime. and this pattern is the basis of freshness grading schemes. so lower carbon dioxide concentrations may have better preservative effects. C (C=reject) Significantly. Spoilage of fish starts upon death due to autoxidation (oxidation of unsaturated lipids). loss of the fresh characteristics may be simply measured by comparative visual and smell analysis. and metabolic activities of microorganisms present in the fish. Degradation of whole fresh fish stored in ice generally follows a set pattern. Over time.60% carbon dioxide with a balance of nitrogen for fatty fish have been recommended (16. Taste panel evaluation of cooked fish may be used to score or grade colour. 1. reactions caused by activities of the fish’s own enzymes. to brown and slimy with an intense off-odour. Loss of freshness and spoilage cannot be separated as processes. TABLE 1. evaluation of taste is unavoidable as the visual pattern of degradation of the eyes and gills is unavailable for analysis. A. Gas mixtures of 40% CO2/30% N2/30% O2 for white fish and 40 .CHILLED AND FROZEN RAW FISH off-odours. When working with fillets. B. or for that matter evaluation of the taste of the fish.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). the gills from pink and shiny. these are summarised in Table 1.III Fish-quality scales Name of scale Torry Scale Quality Index Method European Grading Scheme Scoring system 10 to 0 (≤ 4 reject) 0 to 24 (arbitrary reject) Extra. The eyes turn from convex and clear to concave and opaque.4. but it is a commonly held view that loss of freshness is related to autolytic degradation and spoilage is more microbial in origin (1). as traditional spoilage patterns of the external organs are typically very distinct. smell and 9 . texture. Various methods for whole fish freshness evaluation by a trained panel have been proposed. 18) and are probably the most widely used. and the flesh turns from bright and elastic to dull and soft (1). 17.
rigor resolves. 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. sea-weedy taste. 23). biochemical and microbiological actions. the pH drop of the muscle is correspondingly less. 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.g. After some time. 19). an osmoregulator. By measuring the concentrations of the six components. the cooked analysis of fish passes through four phases of spoilage: i) Delicate sweet. or its remaining shelf life (1). The process of ATP breakdown is used as an indicator of fish freshness (24). RT Freshmeter or Fishtester). During the initial storage. spoiled and putrid. After death. reduction in ATP and increase 10 . a ratio of concentration of the hypoxanthine (Hx) and inosine (HxR) to total concentration (ATP.5 (18. ADP. as little endogenous reduction of TMAO occurs (18). iii) Traces of sour. which has been ascribed a number of possible functions as: a trimethylamine (TMA) detoxified waste product. ii) Neutral taste. the Torrymeter. bacterial reduction of TMAO to TMA. possibly slightly metallic. the muscles harden as residual adenosine triphosphate (ATP) is reduced and the myosin and actin filaments bind to form actomyosin (18). the conductance generally increases. AMP. fruity and/or bitter off-flavours. Because fish tend to have relatively little residual glycogen compared with mammals. To a varying extent. texture becomes soft and watery or hard and dry. an intense odour compound. However. ammoniacal. the muscles relax again and the fish returns to a flaccid state.8 to 6. thus. These processes are strongly influenced by the physical conditions of storage. Generally. 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. which increases from 0% towards 100% with time. IMP. little flavour.FISH AND SEAFOOD taste of cooked fish. development of sickly sweet. iv) Enhancement of the spoilage characteristic of phase iii. cabbage-like. As the fish degrades. an anti-freeze. %). HxR and Hx) gives a quantity (K-value. From being flaccid. or simply a waste product present due to bioaccumulation (22). is significant and may even be responsible for the ultimate sensory rejection of fresh cod and other fish with high initial TMAO content (20. rigor mortis is the first noticeable change in the fish. TMAO permits a high Eh to remain in the muscle tissue. this will generally correlate with the length and severity of rigor. The processes of degradation being analysed by the quality scoring methods above are a complex mix of physical. all marine fish use/have TMAO (21). sulphurous and/or rancid smells. post-rigor values are typically in the range of pH 5. the Eh of fish muscle remains relatively high (20). chemical. After death.
For some fish species. The variation in post mortem fish pH may likewise influence the amount of bases being liberated to the air and consequently affect the odour characteristics of the fish. the majority of volatile bases are produced from the soluble non-protein nitrogen of the fish (free amino acids and other low-molecular-weight nitrogenous compounds). these measurements are usually used to quantify the amount of spoilage. During spoilage. a correlation can be made between the spoilage of the fresh fish and the production of TVB-N.CHILLED AND FROZEN RAW FISH in hypoxanthine by endogenous enzymes allow this measurement to be used for determining freshness (19). for example the TMA fraction. also termed TVN). 27) produced by aerobic respiration and other forms of metal ion reduction (28). Chemical degradation continues after the initial post mortem phase. Volatile bases are the best-characterised chemical indicators of fresh fish spoilage. 18). 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. 26). Evaluation of Total Volatile Base-Nitrogen (TVB-N. however. 7). so the measurement is effective only during the initial loss of freshness. Lipid oxidation and other oxidative changes lead to oxidative rancidity. the residual acid is then titrated. as microbial spoilage is limited by the low storage temperature (1. using Conway diffusion chambers (29) allows determination of changes of mg-N/100 g fish. Comparing results for different fish species. is later significantly reduced by bacterial action. hypoxanthine. does not show correlation between muscle pH and the amount of volatile bases contained within the fish at rejection. as significant proteolysis is observed only during the latest stages of spoilage and after rejection (30. not to describe freshness. however. 31). Oxidative rancidity is known to reduce the quality of fatty fish in particular (19. or a specific fraction of the volatile bases. the importance of microbial action increases with time (5. and are especially important in the spoilage of frozen fish. colour changes. This measure is highly dependent upon the temperature of storage and may not reflect the rate of loss of quality equally at different temperatures. which may also be formed by bacteria (25). However. and a segregated weak acid to absorb them. Neither does the change in pH during storage correlate well with the production of TVB-N. 11 . The major spoilage odours and flavours of fresh fish are undoubtedly principally microbial in origin. therefore. Lipid oxidation can be a result of enzymic action or a cascade reaction initiated by free radicals (26. 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 Conway method (and variations of it) uses a strong inorganic base to volatilise the bases in the fish sample.
the pH of the fish muscle may change (intrinsic) and a different microbial population may develop due to the change in atmosphere and pH. All chemical reaction rate kinetics (and thereby microbial growth) are temperature-dependent.and temperate-water fish spoilage rates may be many. by altering the gas atmosphere (extrinsic). 1. the slower the spoilage processes proceed (within limits). For fresh fish. the lower the temperature of storage. The factors are interactive and cannot be completely isolated. but also crucial for the assurance of consumer safety. This increase has been reported to be moderately dependent on the temperature of the sea from which the fish are taken (33. realisation that the bulk of the microbial population on newly caught fish does not cause off-flavours and off-odours stems from work 12 .5 1. biochemical and microbiological reactions taking place within and on the fish.2 Fish-spoilage bacteria The parameters affecting the multiplication of microorganisms in foods have been categorised into two general groups: intrinsic (inherent qualities of the food) and extrinsic (qualities of the food environment) (35).1 Factors Affecting Fresh Fish Spoilage Temperature By far the most effective method of reducing the rate of whole fresh fish spoilage is temperature control (1.5. Fish spoil as a result of the chemical. For fresh fish iced immediately after capture and continually stored in ice.FISH AND SEAFOOD 1. yoghurt spoilage by yeasts and clostridial spoilage of cheese (5) are examples where it has been recognised for many years that a particular minority of the microbial flora present in the product is responsible for its spoilage.5. The application of ice storage increases the shelf life of fresh fish from a matter of hours at ambient temperatures to days or weeks. a minority are often associated with the majority of the spoilage. including the ability of the endogenous bacteria to grow at low temperatures and lower endogenous enzyme activity. For example. Also. the results of altering one parameter may have far-reaching consequences on others. The reasons for the differences between tropical.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. Not all the bacteria growing on a food will lead to the production of objectionable characteristics. 34).5. The concept of specific spoilage organisms (SSOs) is not new. The sum of these changes may result in a different spoilage profile. careful temperature control during storage is not only important in terms of quality loss. 5). 1.
37). form methylmercaptane (CH3SH) and dimethylsuphide ((CH3)2S) from methionine and produce hypoxanthine (Hx) from inosine monophosphate (IMP) or inosine. It is able to grow as fast as. During the 1970s. putrefaciens to the spoilage of fresh marine fish has been recognised since the 1940s. As previously noted. 36). It is also present in fresh water. although only since the late 1960s has it been realised that specific metabolites of Sh. on fish are diverse and in many cases species-specific. and proteolytic and lipolytic enzymes that may affect the quality of fresh and. mediated spoilage is characterised by ‘fruity’. putrefaciens is considered an SSO of temperate-water marine fish species stored in ice.e.3. more especially. The spoilage compounds associated with the growth of psychrotrophic Pseudomonas spp.5.g. Bacteria identified as being associated with the spoilage process of fresh fish are as follows: 1. Members of the genus are able to produce pigments.2 Shewanella putrefaciens Sh. The spoilage of freshwater fish is generally ascribed to the growth of Pseudomonas spp. 35. putrefaciens growth may be used as indicators of spoilage (30). plus other characteristic compounds of the species responsible for spoilage. Sh. produce hydrogen sulphide (H2S) from cysteine. its ability to reduce TMAO to TMA. and may play some role in the spoilage of freshwater fish. 1. 35). catalase-positive. it is often isolated as about 1 . work with inoculated sterile fish blocks resulted in identification of a specific minority of microorganisms. but only if or when they reach numbers capable of producing sufficient spoilage compounds to effect rejection do they become the SSOs of the product (9. the characterisation of spoilage of each product must be made before the identification of the responsible agent(s) can proceed. It is thus an important spoilage organism of gadoid fish such as cod.5. esters and nonhydrogen sulphide sulphur-containing compounds such as methyl sulphide (36. Pseudomonas spp. which produced the characteristic spoilage compounds of the fish (30. motile with polar flagella. putrefaciens spoilage of fish is due to its biochemical action on muscle. frozen) fish products. i. or faster than the rest of the flora of ice-stored fresh marine fish (9). oxidase-positive. ‘oniony’ and ‘faecal’ odours from the production of ketones.3.10% of the total flora of fresh fish from temperate marine waters.1 Pseudomonas spp. processed (e. aldehydes. They are generally characterised as Gram-negative rods. obligate respiratory bacteria. (9) and they are considered SSO of iced freshwater fish (35). The Pseudomonadaceae family represent a large and poorly defined group of microorganisms. The importance of Sh.CHILLED AND FROZEN RAW FISH started in the 1940s. These organisms are described as potential spoilers. 13 . therefore. the spoilage of a product may be strongly influenced by the conditions under which the product is held.
41). Sh. It is also able to produce hydrogen sulphide and a range of other off-odour compounds. phosphoreum increased in notoriety when it was proposed that reduction of TMAO to TMA limited the shelf life of MAP cod fillets (12). although. putrefaciens was not present in such numbers (23. In fact. The ferric reductase activity of Sh.3 Photobacterium phosphoreum Recognised for some time as being present on spoiling fish (30). was capable of being in a significant numerical minority on the MAP fish but still able to yield the majority of the TMA thought to be responsible for the rejection (23. Shewanella affinis. and recent taxonomic description using modern molecular methods bifurcated this species. they are heterogeneous. The localisation of the reductase on the outer cellular membrane offers the bacteria a solution to this problem (40). mesophilic strains of Sh.how to reduce a molecule that they are not able to take into their cells efficiently.5. unlike many other motile bacteria. In terms of its taxonomy. additional psychrotrophic species such as Shewanalla baltica. as the ferric iron is insoluble. does not show carbon-source chemotaxis. Sh.3. Shewanella waksmanii. Ph. the bacteria have a significant problem to overcome . but Sh. Later. Shewanella olleyana. phosphoreum sufficient to cause spoilage was found. but does show strong chemotaxis up gradients of most of their alternative electron acceptors (39). Shewanella gelidimarina. 38). putrefaciens were required. Ph. and Shewanella aquimarina. baltica has been identified as the main H2S producer in cod during cold storing (37). at the time of rejection. putrefaciens strains are characterised as microaerophiles or anaerobes. with several new species being described. 41). The prevalence of the bacterium in so many environments. phosphoreum were required for 50% of taste panellists to reject a sample. like all members of the Pseudomonadaceae.FISH AND SEAFOOD for which the most compelling evidence showing spoilage as a result of Sh. a population of Ph. 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. This wide-ranging respiratory capability is thought to be unique (39). 1. Shewanella frigidimarina. putrefaciens were identified as Shewanella algae. putrefaciens lies in its chemotaxis: the bacterium. Shewanella oneidensis. putrefaciens may grow in the absence of oxygen using alternative terminal electron acceptors (ATECs). Sh. putrefaciens has been studied. Shewanella livingstonensis. putrefaciens growth has been collected (12. owing to its cell size and activity. In 50% N2/50% CO2 MAP cod. whereas >108 cfu/g Sh. Shewanella denitrificans. and its metal ion reduction ability has led to intensive investigation into its role in iron and sulphur cycles. Research has shown that approximately 107 cfu/g of Ph. it was proposed that this organism. Initially. This type of intoxication occurs when bacteria convert the histidine 14 . phosphoreum has also been identified as responsible for histamine fish poisoning. it is strictly respiratory. Another distinct feature of the physiology of Sh.
6. algal toxins. There are few human bacterial pathogens that can cause primary infections or disease and are capable of persisting in the aquatic environment. Of these.1 Clostridium botulinum C. especially type E botulinum intoxication. age. 75% (with a 35% mortality rate) occurred in Japan. The heterogeneity of the types (A. this is probably due to spoilage occurring before toxin elaboration. 1. Acetate production has been reported as a good indicator of spoilage by this organism. 5) reports that fresh fish consumption has never been shown to cause human botulism. a convenient but diverse species of bacterium with a number of different types.3 °C (Group II) and these are non-proteolytic (i. 1. C1. Types A. bacteria and higher parasites all present some risk associated with fish consumption (1. Growing evidence suggests a role for B. Adult disease is caused by the production of the botulinum toxin.5. a neurotoxin that causes flaccid paralysis and is associated with a variable (44) mortality depending upon dose. B. including antisera. 1. previous exposure and access to supportive treatment. thermosphacta is a well-characterised psychrophilic spoilage organism of meat. B. When inoculated into VP corned beef and sliced ham.CHILLED AND FROZEN RAW FISH present in fish into histamine.4 Brochothrix thermosphacta and lactic acid bacteria B. botulinum recorded up to 1963. but with proper cooking. Some Ph. E and F include bacteria with minimum growth temperatures of approximately 3. D. Of 404 intoxication outbreaks of type E C. MAP studies have also demonstrated this organism’s sensitivity to oxygen and have shown that they are also inhibited by high CO2 concentration (43). thermosphacta in the spoilage of some MAP fish.6 Pathogens: Growth and Survival Endogenous chemicals. types B. F and G) is well documented (44). E.e. Huss (1. Review of outbreak data suggests fresh fish to be safe. phosphoreum strains have great capacity as histamine producers even under refrigeration conditions. presents a potential risk. 6). lightly preserved fish products are associated with botulism. C2. However. thermosphacta on spoiling fish in a 40% CO2/30% N2/30% O2 MAP (42). thermosphacta did not produce off-flavours until 2 . The remaining few bacteria present a major risk involved with the consumption of raw seafood such as sushi or oysters. it is bacterial risks that increase after capture of the fish. E. (and very rarely F and G) have been reported to cause human disease. 5). B.3. Recent studies have investigated the dominance of B. Fewer still are capable of growing on fresh fish. these risks are substantially reduced (1. botulinum.3 days after having reached 108 cfu/g (42). human viruses. 5. where a particular 15 . they do not produce significant product spoilage).
and when in contact with contaminated water (no CDC reported cases from fish. the majority are associated with products not cooked immediately before consumption.6.2005). 5). or cooking at 80 °C for 5 minutes) (44) will substantially reduce risk. were associated with finfish between 1978 and 1998 (46). and North America. whether stored aerobically or in modified-atmospheres. A unique clone of V. Russia.3 °C) will eliminate all risk. 11). 1. V. parahaemolyticus in sea water fish and shellfish that could potentially be hazardous to humans. 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. I-sushi. which is believed to be required for virulence. including epidemics in India. Currently. accounted for the majority of cases (1. parahaemolyticus O3:K6 is responsible for many of the recent V. a Japanese boiled and semi-dried sardine dish. Only about 1% of marine isolates produce a thermostable haemolysin. vulnificus is associated with warm marine and estuarine waters. The bacterium causes primary septicaemia. salted or pickled) fish have also been associated with botulism. The first report of the organism being a foodborne agent was with shirasu. due to the cultural preference for raw fish dishes. Despite the ubiquitous nature of type E C. Generally. parahaemolyticus infections since 1996 and is referred to as the pandemic strain (47). V. which was probably contaminated from an uncooked food source or an excreting foodhandler. is small.FISH AND SEAFOOD fermented raw fish product that is consumed uncooked.g. and proper cooking (boiling for 1 minute. storage and preparation (11). Human disease caused by the organism has only been observed in conjunction with marine bivalve (mostly oysters) and some crustacean consumption. but as fresh fish are often cooked before consumption and the botulinum toxin is heat-labile. again.2 Vibrio parahaemolyticus and other vibrios V. Since 1996. especially in individuals with underlying diseases (patients suffer from immune and liver diseases or blood 16 . USA. This strain has been responsible for 50 to 80% of all V. use of post harvest treatments and/or cooking of fresh seafood will reduce risks. only shellfish are associated with the disease and no reported cases by the Center for Diseases Control. Proper temperature control (<3. 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. parahaemolyticus outbreaks. In Japan. the risk posed to the consumer of fresh fish. parahaemolyticus is a marine organism. the majority of outbreaks are caused by consumption of raw fish products (e. Other lightly or semi-preserved (e. there is no specific guideline that describes a minimum level of V. parahaemolyticus cases have increased across the world. sushi) (5). Southeast Asia. there remains only an ‘extremely small’ risk of intoxication if reasonable precautions are taken during handling.g. The risk of toxin formation before apparent spoilage has been studied for some MAP fish (45). Proper chilling. smoked. France. Japan. 1978 . which can cause human gastroenteritis.
The major virulence factor appears to be the production of toxins with enterotoxic.6. It is particularly associated with disease following consumption of raw oysters from warm sewagepolluted waters. Although the psychrophiles are unable to grow at body temperature. and fish and shellfish are probably the major reservoirs for the organism (50). coli on isolation media (50).6. 17). 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. monocytogenes has been well documented as a foodborne human pathogen. is historically associated with faecally contaminated water.55 cases per 100. There is no risk associated with the consumption of properly chilled and cooked fresh fish. shigelloides is similar to Aeromonas spp. They are aquatic organisms with an epidemiology that is yet to be fully understood. infection. Since 2000. no more than circumstantial evidence exists linking the consumption of seafood to Aeromonas spp. but the bacterium is known to survive and grow in the shallow marine.3 Aeromonas Aeromonas spp. 1.. 5. but proper cooking should substantially reduce risk of infection. Chill storage may not eliminate growth. with minimum growth temperatures around 0 °C and optima of 15 .CHILLED AND FROZEN RAW FISH disorders). V. proper chill storage will eliminate any risk associated with this organism. are associated with human diarrhoeal illnesses. L.20 °C. Despite this. there is no risk from the consumption of properly handled and cooked fresh fish. cholerae O1. are present on fish and can grow to significant numbers during storage (1. sodium channel blocking and haemolytic activity (48). Again. Most cases have been sporadic rather than associated to large outbreaks. 1. and especially estuarine environment. in its habitat. others are psychrotrophic mesophiles (49). Similar problems as those encountered for Aeromonas spp. The bacterium has been documented as being responsible for a few fishborne outbreaks of gastroenteritis. the causative agent of cholera. which is often fatal (>50%).26 to 0. though it is a true mesophile with a minimum growth temperature of 8 °C and shows consequential seasonal variation in its environmental isolation. listeriosis cases have been reportable to the CDC. Some isolates are true psychrophiles.000 17 . in assessing the risk of fishborne infection by P. cytotoxic. It is known that Aeromonas spp. possibly due to under-reporting because of their likeness to E. monocytogenes incidence has been between 0. and especially Aeromonas hydrophila. their potential to produce disease-causing exotoxin in food has not been fully examined (48). and proper cooking will substantially reduce risk.4 Listeria monocytogenes L. however. shigelloides are encountered. In the US. P.
It is environmentally ubiquitous but its true frequency in the marine environment is not well studied. The organism poses a serious risk in chilled products not cooked before consumption. alcoholics. and Lactobacillus spp. especially pickled fish. it is not apparent to the consumer and it cannot be destroyed by cooking. AIDS patients and patients undergoing immunosuppressive therapy) and a lack of data covering infectious doses.FISH AND SEAFOOD persons. If fish are not properly refrigerated. but proper cooking will reduce any risk. there is a far greater risk of unacceptable histamine levels being attained before sensory rejection. 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.5 Scombroid fish poisoning The production of histamine and other biogenic amines with human immunological activity. D60 of 1... Infections are mostly associated with the consumption of raw or 18 . 1. within high histidine-containing fish (especially members of the Scombridae and Scomberesocidae families) is responsible for an intoxication termed scombroid poisoning (54). some Vibrio spp.6. scombroid poisoning is usually a mild intoxication and it is not a significant cause of death. Like other toxins. but more especially by Morganella morganii. Of fish-transmitted human diseases scombroid poisoning is commonly reported in the US (118 outbreaks with 463 cases reported by CDC 1998 .48 minutes in salmon have been reported (53). there is a possibility for growth of the organism on chill-stored fish. Lightly preserved products. 1. 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.6. neonates. Klebsiella pneumoniae and Hafnia alvei. Photobacterium spp. The most frequently reported parasites of human importance in fish are round worms of the genera Anisakis and Pseudoterranova and tapeworms of the genus Diphyllobothrium. Fortunately.2002 (55)) and is also common worldwide. Low-temperature storage of potentially toxigenic (<5 °C) fish at all times is the most effective way to control histamine production (6).pregnant women. at risk groups .6 Parasites Helminthic parasites can occur extensively in finfish but very few are capable of infecting humans. foetuses. leads to the production of histamine. It is regularly isolated from seafoods (51). are exceedingly difficult to produce within the legal limits of histamine as the temperatures used during production can lead to rapid growth of histidine decarboxylating bacteria.. Owing to the dependence of the virulence of this organism on the immune state of the host (usually requiring lowered cellular immunity. full risk analysis is not possible (52). Decarboxylation of histidine by a wide range of bacteria including Enterobacteriacae.98 minutes in cod and D60 of 4. Clostridium spp. Being a psychrotroph. but cooking will significantly reduce the risk of infection.
Testing methods should be practical and the enforcement of such criteria should translate as a reduction of potential microbiological risks to consumers. 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.1997. including parasites and/or quantity of their toxins / metabolites per unit of mass. risk of human disease caused by natural bacterial contamination (not through faecal pollution) of fresh fish is extremely low. The most effective control of all bacterial risks is continual chill storage in ice (0 °C) or freezing.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. 444. coli Case 1 4 Plan 3 3 n 5 5 c 3 3 m 5 x 105 11 M 107 500 Additional tests to be carried out when appropriate Salmonella V. parahaemolyticus Staph. 2003.. An increase of APC to levels in excess of 106 cfu/g is usually indicative of inadequate refrigeration. especially in warm-water areas and where fish are to be consumed raw. it may be desirable to test for Salmonella and V. No. area or lot (CAC. parahaemolyticus (see Table 1.CHILLED AND FROZEN RAW FISH mildly processed fish such as sushi. Cooking properly immediately before consumption is also an effective way of reducing or eliminating risk of fresh fishborne disease. Ababouch L. For fish from inshore or inland waters of doubtful microbiological quality. In summary. 1997) Huss H. Rome. FAO. Faecal coliform counts may be used instead of E. Criteria should set standards that are attainable by the currently accepted GMP and applied only when there is absolute need for it. which in many cases will eliminate the risk. EC.H. 1. Assessment and management of seafood safety and quality FAO Fisheries Technical Paper.. coli counts where this method is preferred.IV) (57). TABLE1. A limited number of nematodes and trematodes found in finfish have also been identified as a cause of human disease (56). Gram L.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. long storage under refrigeration or one of the former prior to freezing. coli proposed by the International Commission on Microbiological Specifications for Foods (ICMSF). aureus 10 7 7 2 3 3 19 5 5 5 0 2 2 0 102 103 103 104 . volume.
107-27. Incidence and growth of psychrotrophic Clostridium botulinum in foods.A..H. Food and Agriculture Organisation. Food and Agriculture Organisation.K. Simpson B. 3. Rosnes J. 283-94. 5. 2001. Kirby R. Incidence of foodborne pathogens in European fish. Rome. International Journal of Food Microbiology.J. Davey K. Spoilage and shelf life of cod fillets packed in vacuum or modified atmospheres.. 334. 19. Armstrong D.. 1651-60. Microbiological spoilage of fish and fish products.R. International Journal of Food Sciences and Techonology. 5-7.S. International Journal of Food Microbiology.A. in FAO Fisheries Technical Paper No. Davies A. 2002. 4.P. Dalgaard P. Rome. Jehanno D. 87121.. 291-304. Huss H.E. Capell C. in FAO Fisheries Technical Paper No..H. 12.J. 2.. Critical Reviews in Food Science and Nutrition. Food Control. 2.H. Sivertsvik M. 1994. 11.R. Jeksrud W. Food and Agriculture Organisation of the United Nations. 67-71. 12. Reddy N. 1992.E.N. 9. The role of seafood in bacterial foodborne diseases. Microbes and Infection. Codex Alimentarius Commission. activities and safety. 24.. Codex Alimentarius Commission.R. Huss H.. 13. Review: Assessing and managing risk due to consumption of seafood contaminated with microorganisms. Shelf life extension and safety concerns about fresh fishery products packaged under modified atmospheres: a review. Smith J.8 1. 1992. and natural toxins.. Ashie I.R. Kautter D. 14. 2003. 91-8..FISH AND SEAFOOD 1.. 348. Italy. A review of modified atmosphere packaging of fish and fishery products – significance of microbial growth. 1993. Davies A.. Huss H. 33. Ed. Gram L.A. 1996. Food and Agriculture Organisation of the United Nations. M. Control of indigenous pathogenic bacterial in seafood.K. 1994.. 36. 1989. Journal of Food Safety. 87-118. 8. 1995. 6. Ed. Food Control. 121-37. Spoilage and shelf life extension of fresh fish and shell-fish. 20 .. 37.T. Fletcher R. Rhodehamel E. Theoretical analysis of two hot water cabinet systems for decontamination of sides of beef.H. Food Control. 10. 243-60. Nychas G. Gibbs P. References Huss H. 2000. 8 (2). International Journal of Food Microbiology. Huss H. Feldhusen F..H. Quality and quality changes in fresh fish. 1997. Ahmed F. Code of Practice for Fish and Fishery Products (CAC/RCP 52-2003). Assurance of seafood quality. 5 (1). Gram L.. 27.J. 12. 1996. parasites. 7. International Journal of Food Science and Technology.
J. 17. Conway E. Huss H.spoilage of fish. 121-37.. 34. Hebard C. The micro-determination of ammonia. Ed. 1415. 354-8. National Advisory Committee on Microbiological Criteria for Foods.. Martin R.. Journal of Applied Bacteriology. 305-17.ucdavis. Non-enzymatic oxidation of seafood lipids. Ratnayake W. 1992. http://seafood. 117-27. 21.E. Børresen T. 33.N.H. 1995. Schwall D.M.J. 33. Part I . 26. Fate of Aeromonas and Yersinia on modified-atmospherepackaged (MAP) cod and trout. Rome. Biochemical basis of post-mortem nucleotide catabolism in cod (Gadus morhua) and its relationship to spoilage. Eds.J. National Advisory Committee on Microbiological Criteria for Foods.J. International Journal of Food Microbiology. Khayat A. Qualitative and quantitative characterisation of spoilage bacteria from packed fish. Ed.. 42-54. 11. 184. Huss H. Quality and Quality Changes. Byrne A.. Eds. Arai K. 1933. Nature.E. Quality and Quality Changes. Food and Agricultural Organisation of the United Nations.G. 1995. 41-50. 19. Flick G. Journal of Food Quality. Occurrence and significance of trimethylamine oxide and its derivatives in fish and shellfish. 419-29. Gram L.. Hebard C.E.. Rome. Martin R.edu/haccp/compendium/chapt08.M. 29.A. International Journal of Food Microbiology.E..J.CHILLED AND FROZEN RAW FISH 15. in Fresh Fish.htm Anon. Lipid oxidation in seafood. 23. 20..A. in Advances in Seafood Biochemistry. Autolytic changes in fish post mortem. 19-22.J. Martin R. 36. March 20. 130-2. 1996. Leblanc P... Ackman R.M. 37. Food Technology. Flick G.. Dalgaard P. 1982... 27. Surette M. 1987. Chemical composition. 1983. Herbert R. 1995. 1-18. 26. Huss H. Gill T. Guidelines for the Handling of Fish Packed in a Controlled Atmosphere. Flick G. Sea Fish Industry Authority.. 1985. Washington D. 1959. . Shewan J. Vacuum or Modified Atmosphere Packaging for Refrigerated Raw Fishery Products.M.E. Hendrie M.J. 22-36. 1992.J. Edinburgh. Saito T. Lancaster. Gibson D.E. Yajima T. 30. Huis in't Veld J. 27.. Microbial and biochemical spoilage of foods: an overview. Shelf life extension of fresh fish .R. 24. AVI Publishing. 1. Microbiological spoilage of fish and fish products. 1971. 22.C.H. in Chemistry and Biology of Marine Food Products. Journal of Agricultural and Food Chemistry. Technomic. Changes in purine nucleotide of red lateral muscle of rainbow trout. Reginstein J. 25. 28. 1995. 1988. Slade A. Adopted.. International Journal of Food Microbiology. in Fresh Fish. Biochemical Journal. Bacteria active in the spoilage of certain seafoods.S. Food and Agricultural Organisation of the United Nations. Gill T.a review. 1996.H. Davies A. 245-67. Letters in Applied Microbiology. 2007. 18. Pedrosa-Menabrito A. Westport. 21 16.H. An absorption apparatus for the micro-determination of certain volatile substances.. 21.
1992. 33. 1994. 109-36. Identification of Shewanella baltica as the most important H2S-producing species during iced storage of Danish marine fish. 1984.. Do Tropical Fish Keep Longer in Ice Than Temperate Fish: The Circumstantial and Definitive Approaches..eu/LexUriServ/LexUriServ. Environmental Biotechnology. 36.H. The inhibition by CO2 of the growth and metabolism of microorganisms. 36. 1989. COUNCIL REGULATION (EC) No 2406/96 of 26 November 1996 laying down common marketing standards for certain fishery products http://eurlex. a dominant microorganism in Mediterranean fresh fish (Sparus aurata) stored under modified atmosphere... 9. Rome. 311-43. 48. 22 . 1982. 317. Gorczyca E. Annual Review of Microbiology. Venkateswaran K.. Localisation of cytochromes to the outer membrane of anaerobically grown Shewanella putrefaciens. 43.. Rome. Satomi M. 6689-97. Archiv für Lebensmittelhygiene. 1989. Ed. 42. Scanlan R.. 32. and regulation. 323-9. Miller A.B. II. Karnop G. Processes in Pathology and Microbiology. Journal of Bacteriology. 38. FAO Fisheries Report.europa. in Fresh Fish. Sumner J. 35. Brochothrix thermosphacta.J. Blackwell Scientific Publications. 1995. Applied and Environmental Microbiology.. The effect of storage temperature on shelf life.H.F. Italian Journal of Food Science. Huss H. Gram L. 174.M. Libbey L. Huss H. Drosinos E. Dixon N. 37. 328-9.H. Lee J.A. 41. 39. Dalgaard P. 33. 34.H. 2005..L.J. 72-80.R.M. Dalgaard P.. 26.. Fish spoilage bacteria – problems and solutions. Brady P. Iron and manganese in anaerobic respiration: environmental significance. Vogel B. International Journal of Food Microbiology.E.. Food and Agricultural Organisation of the United Nations. 44. Taussig M. Nychas G. 62-70.R. 4. Do fish from tropical waters spoil less rapidly in ice than fish from temperate waters? Food Technology in Australia. 2002. 40. Kell D.do?uri=CONSLEG:1996R2406:20050602:E N:PDF Sumner J. Myers J. 1996. 61-6. Magno-Orejana F. Oxford. 334. Journal of Applied Bacteriology. Myers C. Nealson K. Jørgensen B. Volatile compounds produced in sterile fish muscle (Sebastes melanops) by Pseudomonas putrefaciens.M. Gram L. Applied Microbiology. 1973. 13. 51-62. physiology. Saffarini D. 262-6. and an Achromobacter species.. 3429-38. 18-21.FISH AND SEAFOOD 31.S. Quality and Quality Changes.. Growth and activity of Shewanella putrefaciens isolated from spoiling fish.. Cohen D. 1984. 67. Occurrence and importance of proteolytic bacteria as spoilage indicators.. 71. Role of proteolytic bacteria in fish spoilage. Pseudomonas fluorescens. 1985.
L. USA. Embarek B. Embarek B. Garcia G. Presence. 20. 49. PCR detection of a newly emerged pandemic Vibrio parahaemolyticus O3:K6 pathogen in pure cultures and seeded waters from the Gulf of Mexico.J. 303-12. Use of risk assessment to reduce Listeriosis incidence.. 17-34. New York. Smith J. 181-96.. Daniels N. Food Technology. 47. 330-6.K. Experimental evidence for toxin production by Aeromonas hydrophila and Aeromonas sobria in a meat extract at low temperatures. 2000.. Golden D. Myers M.. 1986.. 2000. Loessner M. Ljungh A. 23 . 1998-2002. Microorganisms in Foods 2. Ray B. 2194200.P. Journal of Food Protection. 1-37. Hammond R. Modern Food Microbiology. 1997.. Panicker G. Thompson S. Whiting R. Surveillance for Foodborne-Disease Outbreaks .. Blackwell Scientific Publications..J. 52. 69. 2003. Slutsker L.W.. Olley J. 172-249. Oxford. 2005. International Journal of Food Microbiology. 1994. Huss H. 85-95.. 48.. Fish and Fish Products (Microorganisms in Fish Products). Miller A. MacKinnon L. International Journal of Food Microbiology. 50.A. 53. 2006. Jay J. Braden C.H. Griffin P.. Kluwer Academic/Plenum Publishers. 100-3. 50. 1987.. E and F in salmon fillets stored under modified atmospheres at low and abused temperatures. Journal of Infectious Diseases. 1991.. Vibrio parahaemolyticus Infections in the United States.. Heat resistence of Listeria monocytogenes in vacuum packaged pasteurised fish fillets.. Histamine fish poisoning revisited.L. Painter J. 57. 12. 54. 1-34. Aeromonas and Plesiomonas as food.. The International Commission on Microbiological Specifications for Foods. Lynch M.C. Genigeorgis C. Applied and Environmental Microbiology. International Journal of Food Microbiology. 12.A.M. 46. Bean N. Sampling for Microbiological Analysis: Principles and Specific Application.. Springer. 51 (4). The International Commission on Microbiological Specifications for Foods. in Microorganisms in Foods 6. The International Commission on Microbiological Specifications for Foods. 58. Lehane L.. Microbial Ecology of Food Commodities. Mac Rae I. detection and growth of Listeria monocytogenes in seafoods: a review. 51.United States. 1661-6..P. Woodruff R. 181.H. 2005. Bej A. Bishop R. 181-8. International Journal of Food Microbiology.. Morbidity and Mortality Weekly Report.and waterborne pathogens.N. Lindroth S. Altekruse S.K. Ed.K. Wadström T. Wilson S. 23. 1991. Risk of growth and toxin production by Clostridium botulinum non-proteolytic types B.M.C. International Journal of Food Microbiology. 1993... 55.. Majeed K. 56.CHILLED AND FROZEN RAW FISH 45.M. 55 (SS10). 1973-1998..
in Fish as Food.. in FAO Fisheries Report No.. 35. Fisheries and Food. Sumner J. International Journal of Food Microbiology. Kitts D... Handling and processing of scad. 295-307.M. Ravn Jørgensen B. 317 (Suppl. Picard G. Ogden I.FISH AND SEAFOOD 1. 51. Labuza T.1983. Aberdeen. Cann D. Huss H. Torry Research Station.. Technomic Publishers. International Journal of Food Microbiology.D. Gibson D.C.1 Further Reading Processing Anon..P. Villemure G. Shinano H. 271-85.9. Ed. 1985. 317-21.. 15. in Seafood Science and Technology. Huss H. Food and Agriculture Organisation. 65-72. International Journal of Food Microbiology. Studies on the microbiological ecology of mackerel stored by the method of partial freezing. Detection of specific spoilage bacteria from fish stored at low (0 °C) and high (20 °C) temperatures.H. Prediction of shelf life and safety of minimally processed CAP/MAP chilled foods: a review. The Microbiology of Seawater Fish. McMeekin T. Dalgaard P.L. Taoukis P. Gibson D. 1989. 1984. 1989.. 741-50. Borgstrom G. Application of predictive microbiology to assure the quality and safety of fish and fish products. Torry Research Station of the Ministry of Agriculture.9. Do Tropical Fish Keep Longer in Ice than Temperate Fish: the Circumstantial and Definitive Approaches.E.. 24 . 1988..D. Lancaster. 62-70. Trolle G.H. Rome. 1. Cho Y. Food and Agriculture Organisation of the United Nations.. Magno-Orejana F. Fisheries and Food.2 Spoilage Shewan J. Smith G. 1992.. Olley J. Mathematical Modelling Used for Evaluation and Prediction of Microbial Fish Spoilage. 13-32. Academic Press. 1961. Torry Research Station of the Ministry of Agriculture. Huss H. Simard R.S. 1984. Changes in microflora and chemical compounds in mackerel stored by partial freezing. Hobbs G. Journal of Food Protection.H. 1986.). 1992. Further Studies on Marine Fish Stored Under Modified Atmosphere Packaging. Ed.. 6. Houston N. Microbiological quality and shelf life prediction of chilled fish. Journal of Food Science..M.A. Jones Y. Florida..9 1.G. 1. 55. Eds. 1997. Estimation of the bacteriological quality of fish by automated conductance measurements. Akiba M. 1. Ross T. International Journal of Food Microbiology. in Torry Advisory Note No 93... 127-34..M. Gram L. 73-89... Shahidi F. Bulk storage of cod fillets and gutted cod (Gadus morhua) under carbon dioxide atmosphere. Aberdeen. Bulletin of the Faculty of Fisheries (Hokkaido University). 4.
H.. 1992. Blackie Academic & Professional. Eds. 1. International Journal of Food Microbiology. Inhibitory effect against pathogenic and spoilage bacteria of Pseudomonas strains isolated from spoiled and fresh fish. 1. 4. 1987. Jacobsen M. 479-88. Trolle G. 1993. Ed. pre-incubation at low temperature and pH on the thermal resistance of Aeromonas hydrophila.europa. Applied and Environmental Microbiology. Gram L. Gram L. in Quality Assurance in the Fish Industry..H. 322-6.3 Pathogens Condon S.M.. Elsevier Science.. 65-72.9... 5. 137-46. London. 1987. Detection of specific spoilage bacteria from fish stored at low (0 °C) and high (20 °C) temperatures. Hall G. 2197-203. 72.K. in Fish processing technology. Otero A. Garcia M. 25 . Huss H. Effect of culture age. 1992.4 Standards and criteria Council Regulation (EC) Number 2406/96 of 26 November 1996 laying down common marketing standards for certain fishery products.do?uri=CONSLEG:1996R2406: 20050602:EN:PDF Davies A.R. http://eur-lex.CHILLED AND FROZEN RAW FISH Einarsson H. 20023. Sala F.J.. Modified atmosphere packaging of fish and fish products. Huss H. 1997.eu/LexUriServ/LexUriServ. Liston J. A rapid method for detection of histamine producing bacteria. Journal of Applied Bacteriology. Amsterdam. International Journal of Food Microbiology.. Klausen N.9.L. Huss H. 59.H. Predicting the shelf life of cod (Gadus morhua) fillets stored in air and modified atmosphere at temperatures between -4 °C and +16 °C.
Sushimi tuna). clarity of eyes.g. CHILLED AND FROZEN PREPARED FISH PRODUCTS Dr. 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. Simon Derrick The Grimsby Institute Humber Seafood Institute Europarc Grimsby DN37 9TZ United Kingdom 2.g. The majority of seafood products produced require a cooking stage before consumption. although recent developments in improved supply chains. Few seafood products are sold ‘ready-to-eat’. and crustacea such as shrimp and crabs. Even though heating prior to consumption will eliminate pathogenic microbes. are used in chilled and frozen prepared meals since many of the attributes on which quality is assessed are not present or noticeable (e. emphasising the high quality of the seafood and the gourmet nature of the prepared products. colourings etc. smoked (cold) salmon.).1 Introduction It was often thought that whilst the highest quality seafood products are processed and sold as whole fish and/ or fresh fillets (e. 27 . smell etc. The same used to be said for the difference between fresh and frozen products. it does not necessarily apply to the toxins that may have been produced. the exceptions being Sushimi (high quality raw fish). This is no longer the case with many processed products now available in stores. due to the early cessation of spoilage. or could be disguised by sauces. seafood (with the exception of some crustaceans) remains uncooked until the point of consumption. 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. colour of gills. smoked (hot) mackerel. products of lesser quality.2. 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. but still acceptable and meeting all safety criteria.
Nevertheless. Frozen fillets may have a surface layer of 28 . Microbiological hazards include Salmonella. Doubts have been raised over the microbiological status and handling practices of reef-caught fish.. At the same time. Cold-water fish microbiological specifications are inappropriate for assessing freshness of warm-water fish. in particular. They are typically flown into export markets. or in MAP depends on catch conditions and treatment.2 Definitions The traditional focus of the chilled and frozen fish market has shifted over the last few years. 2. This includes all categories of seafood including trout and other freshwater species. prawns. they are usually supplied filleted with head on. There is also a limited market for filleted material. They can be with or without skin and may contain pin-bones although the major skeletal bones are removed during filleting. which can be farmed. and pathogenic Vibrio spp. The market spans fresh fish counter sales and frozen recipe dishes with fish as the predominant source of protein.. from both wild capture and aquaculture sources. Listeria spp. they are usually of a better standard than species such as tilapia. and are well known for their diet consisting of waste materials. Wet or chilled/frozen whole fish are whole fresh fish that may or may not have been filleted. Seafood is the term used descriptively to include all categories of product.FISH AND SEAFOOD Recent trends in the market for food and seafood products. as they tend to lose colour and sheen after defrosting. the processing industry has developed a wide range of value added products. including freshwater fish. and the consumer only has to heat (cook) the product. Warm-water/reef-caught fish were initially supplied to add colour and variety to wet fish displays. and this has produced a greater availability and diversity of products in the marketplace. where the preparation has already been carried out. have shown the increasing importance of convenience foods. rather than frozen. Marketing-style descriptions and process definitions are presented below to acquaint the reader with the variety and forms of seafood available. and it is worth noting the additional hazard to fish handlers from Vibrio vulnificus from injuries received during handling and filleting of spiny tropical fish. but these are now finding favour as products in their own right. Prepared natural fillets are fillets of fish without any additional coating. It is sold from wet fish counters on ice or pre-packed in modified atmosphere packaging (MAP). often using the simplicity of a microwave oven. and these are increasingly being presented with an ever-expanding range of accompaniments. etc. Shelf life on ice. but should be equivalent to cold-water products. molluscs. shellfish and molluscs. The expected shelf life attainable for this format is usually up to 8 days.
sawdust with other woods adjusted for flavour profile and burn duration). A core temperature of around 27 30 ºC for approximately 100 minutes is a typical processing condition. Hot-smoked fish are prepared from brined. natural fillets of fish such as herring. 29 . or pre-packed in MAP or vacuum packaging (VP). swordfish steaks). resulting in a product that is ready-to-eat without any further cooking. The pâtés are sold from segregated ‘wet fish’ counters as a ready-to-eat product. These products are packaged in separate high-care facilities to minimise contamination with microbes either from the workforce or from raw products. which is approximately 30% (or 1 . mackerel. tuna steaks) or band-sawn from a frozen block (e. cod). and the product then shaped and heat-processed to cook the product before being chilled. 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. A typical cold smoking process may pretreat the fillet by dipping in a 26% brine solution with a retention time of approximately 3 minutes.CHILLED AND FROZEN PREPARED FISH PRODUCTS frozen water that forms a protective glaze.5% in the aqueous phase. haddock. annatto with smoked haddock. 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. Added colours can be included with the brine solution to meet specific requirements for products e. VP products) because of the high salt content acting as a barrier to spoilage. They are then kilnsmoked to a minimum core temperature of 66 ºC for 2 minutes. 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. mackerel. in addition. the ingredients premixed. The shelf life for MAP fillets is typically up to 5 days. which may.2. or pre-packed frozen with added glaze.2 days) longer than that of non-prepacked fish. Again.g.5%. Fish steaks can be cut from a natural fillet or loin (e. with a minimum residual salt content of 3. They are sold from wet fish counters. and salmon. Cold-smoked natural fillets are prepared from a range of cold water fish including herring. salmon.g. Cold smoking is essentially a flavouring process. The expected shelf life of these products is up to 8 days (or longer for some chilled.5 . be flavoured with herb dips or peppercorns. a protective glaze may be added to the frozen steak.g.g.. Pâtés are prepared with raw fish. The result is fish with a typical residual salt content of 1. etc. This process in effect cooks the product and reduces or eliminates microbial contaminants. in VP or through mail order. Some products may be salted for flavour only (e. Fresh fillets are sold from wet fish counters. but may be increased (up to 1 month) by packaging methods like MAP or VP. and typically uses natural smoke passing over the fillets suspended on racks in a kiln (oak chippings.
Band-saw dust and fish trimmings from fishblock preparation materials may be used. With further processing. and formed into round cake shapes. flavours. salmon) coated and sold as chilled products. rice or pasta. Other. This ‘catch-all’ category includes seafood salads. tuna.) Raw material is typically fish fillet. 30 . it can be used as a seafood analogue. binders. which may include molluscs. jellied eels. frozen blocks or canned fish and may include molluscs or prawns that may or may not have undergone a minimal heat process. long-life seafood mixes using sorbate and benzoate as preservatives. e. They are pasteurised products and treated as high risk raw materials.g. Scampi are tails of Dublin Bay prawn or langoustine. ‘fingers’ before being coated in batter and/ or breadcrumbs. mixed whitefish. etc. coated in batter or breadcrumbs. and seafood mixes. Surimi is the insoluble protein fraction of fish after washing and processing. taramasalata. haddock. The protein source can be fish fillet or blocks. cephalopods.g. The cakes are usually coated using a three.. and heat-fixed. with an expected shelf life of around 7 days. or cut from a preformed frozen fish blocks into shapes. pasta or potato. Breaded fillets/steaks and battered fillets are either natural or shaped fillets.or a five-stage process. and may include molluscs or prawns that may or may not have undergone a minimal heat process during their primary processing. addition of ingredients such as colours. It is often used as a protein supplement. fish nuggets/bites with dips and seafood-based snack products. There is an increasing market for high value fresh or thawed fish (e. Fish cakes are products produced by mixing fish with potato and herbs. Other ingredients may be natural or reconstituted. rice. Prawns. Natural prawns from cold or warm waters can include those peeled. The majority are products made from various types of whitefish (cod.FISH AND SEAFOOD Seafood recipe meal centres (frozen or chilled) are places where these products are supplied without a source of carbohydrate. e. 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 may also include additional ingredients such as fresh herbs and spices. and in the shell.g.
it may be used for other products or markets depending on the product specification.3.e. A primary role in fish spoilage has been assigned to this organism. Contamination will. as a result there is an increase in spoilage rate thereby reducing the product shelf life.CHILLED AND FROZEN PREPARED FISH PRODUCTS 2. see Table 2. The Food and Drug Administration (FDA) has produced guidelines for handling and processing of tuna. The concern is that fish may be visibly acceptable but any subsequent washing process will have little effect on removing contaminants. After capture and during any subsequent processing.1 Initial Microflora Seafood Other chapters have discussed in some detail the indigenous spoilage. In summary. 31 . and his co-workers have described a large celled microorganism found in spoiling fish and identified it as Photobacterium phosphoreum (3). and intestines of the fish. which may become a focus of contamination deep within the muscle. species (Bluefin>Bigeye>Yellowfin>Albacore). the microbial flora of seafood is associated with the gills. skin. and the potentially toxic by-products of histamine-producing bacteria (5. fat content. Tuna that does not meet the highest of grades based on colour. remain on the surface of the fish fillet except where “gaping” or separation of the muscle blocks occurs. Dalgaard et al. whilst gaping is also dependent on a range of other factors that effect the muscle composition. 6). will the saprophytic flora start to invade the surface flesh (1. i. including seasonal variation. storage. Tuna caught and discarded as unsuitable for sushi or other high-risk uses is often diverted for canning. Only after death. For a summary of background information on the hazards associated with fisheries products and ingredients. Recent public opinion has favoured the use of dolphin.and turtlefriendly long lining to the less discriminatory netting. and have emphasised the origin and sustainability of the fish as key marketing points. The edible tissue of some species of fish are more susceptible to gaping than others. on the whole.I. and pathogenic flora associated with seafood. and ingredients added to the final product. and processing conditions such as dehydration or poor filleting practices. handling or preparation stage. is not used for sushi or other high risk (and high value) products. sensory assessment of freshness etc. product handlers. and an increased risk from a potentially unsafe product (especially those that receive a minimal heat process. specifying internal temperature limits for fish and for limiting temperature abuse during handling and grading (1). Grading is based on size of fish (>25 kg). Frequent rebaiting and prompt removal of caught fish is essential to limit rapid spoilage. other potentially pathogenic and spoilage bacterial contaminants may be introduced from a wide range of sources including the environment.3 2. searing or bar marking). 2). or visible damage from gaff marks (4).
nuts and fruit Associated microflora Lactobacilli. Vibrio cholerae. Bacillus cereus. Aspergillus. Listeria. cream. streptococci. TABLE 2. cereus B. Clostridium perfringens. Bacillus subtilis B. and so are the potential microbial contaminants associated with them. yeasts Pathogenic microflora/toxins Salmonella. cereus Bacillus licheniformis. be considered at an early stage during new product development.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. and associated spoilage and pathogenic microorganisms.4. 32 . for chilled products.2 Non-seafood ingredients The range of ingredients used in the preparation of chilled and frozen prepared seafood products is very diverse. by the HACCP team or by a hazard and risk assessment programme. moulds. Penicillium. Clostridium botulinum. 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. cereus Staphylococcus aureus. Moulds Enterobacteriaceae Salmonella. enteric viruses 2. histamine Cryptosporidium. moulds Bacillus. shelf life determination. 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.4 2. raw materials. product assembly. anaerobic spore-formers. yeasts Lactobacilli. Staph. Staphylococcus. Table 2. Escherichia coli.FISH AND SEAFOOD 2. Aeromonas.I provides an overview of common raw materials. B. moulds Bacillus Bacillus. staphylococcal toxin. Pseudomonas. mushrooms.3. and. coryneforms. Enterobacteriaceae. and milk Cheese Water/Ice Bacillus. In addition. Fusarium. toxin. spore-formers. however. coliforms. Clostridium Lactobacilli. Shigella. pseudomonads. intermediate processing. moulds Bacillus. cereus Carbohydrates Potato flake and powder Rice Pasta Flour Dried herbs and spices Dairy products including butter.1 Processing and its Effects on the Microflora Selection of raw materials/ingredients It is important that the potential microbial contaminants of all ingredients. protozoa and viruses B.
5 illustrate basic principles in the preparation of chilled and frozen recipe dishes. as the full complexity of the processes used for specific products cannot be covered in a general text.3.4.4. Generic examples have been used to illustrate only the major points. but it is assumed that any band sawing or dice process will introduce heat from blade friction. Intermediate processing and use of chill for storage of work-in-progress will help to optimise available life of perishable components. to evenly raise the temperature of the entire frozen product from -20 °C or lower. 2. All have the same intention. but it also poses a risk of cross-contaminating other products undergoing tempering. there are a number of distinct methods used. Examples of tempering techniques include the use of heated air. but will also need to ensure that the optimum shelf life can be safely obtained. Effects on the microflora are minimal during tempering (7). Where the raw materials are frozen.2 Raw material tempering Whenever frozen seafood is used as the raw material it is normally tempered before processing.1 .3 Preparation of fish component of chilled and frozen recipe dishes 2. 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.2. Figures 2. Rapid controlled cooling of heat-processed components will help to minimise spoilage. 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. this may be an incidental effect. It is recommended that the principles of HACCP are applied to novel processes. defined holding time at a chill temperature (0 – 3 °C). The use of water for rapid thawing of product is not recommended since it not only rapidly thaws the product surface. and contamination from equipment to clean fish muscle. 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. the core temperature.4. or the use of microwave technology. depending on the size of the fillet 33 . line extensions and process reviews.1 Coated products Seafood products are coated with a cereal-based batter or crumb with up to 30 50 % of the final product weight accounted for by the batter or crumb. 2. up to -5 °C so as to improve the cutting and handling properties during processing. Band-sawn materials or fish dice are normally promptly further processed. as a heat process is often required for functional or organoleptic reasons. For some components.
Shaped Fillets. Individual Quick Frozen (IQF) fish fillets or steaks may also be coated using a similar process.FISH AND SEAFOOD processed.3.1C . 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. Frozen cores are usually batter-enrobed. drained and then refrozen for storage until required for further processing.2).1A .1 Whole-fillet / scampi Scampi (Nephrops norvegicus).1. 2. The processing of three examples types are described in Figure 2. frozen in a nitrogen blast freezer and weighed into sacks for later processing. The scampi meat (tails) is removed intact from the shell using potable water whilst still frozen.4. The “blown” meat is graded.Scampi. chilled or sold chilled on defrost with an expected shelf life of 6 .1B .2. On arrival at the processing plant. It is then size-graded.4. quickfrozen. may not rise above 0 ºC during processing.7 days. Any broken pieces or fragments of frozen tail meat are kept for use in the production of a reformed scampi product. with the coating heat-fixed.1 (2.Fish Cakes).1. Raw material preparation stages for chilled or frozen coated fish products 34 .3. after landing. Fig. the scampi is washed in potable water to remove foreign bodies. by mincing and mixing with a binding agent (as described in section 2. 2. 2. are placed into plastic boxes and covered in ice to preserve their condition.
or sold as a chilled product after thawing. unless it is poorly handled. additional spoilage organisms may be introduced. Herbs or seasonings used in the recipe may contribute additional spoilage organisms or. Fish raw material can be obtained from a number of sources i) By-product of fish shaping or cutting/dicing process . It is for this reason that all by-products from primary processing. are handled and stored as any other food product.2 Shaped products This type of product falls into two categories. from potato flake. such as aerobic spore formers and Staphylococcus spp. quickfrozen. Spoilage of fillet pieces or trimmings will not be appreciably different from that of fillet. because of its nature. and water is added to rehydrate the components. ii) Fish-filleting waste . iii) Fresh/fillet/minced fish block. The mix is shaped and formed ready for packing or for enrobing. that are to be reused for other foods. This type of material will be susceptible to contamination by Enterobacteriaceae and possibly Staphylococcus spp.. The remaining dried ingredients are mixed with the fish.block-sawn dust. Waste material produced from this type of processing is often reused in the third category (fish cakes). 35 . once the dry components have been rehydrated.off-cuts or belly flaps.1.CHILLED AND FROZEN PREPARED FISH PRODUCTS 2. but.3. This is especially the case with block-sawn dust.4. Therefore. or promptly processed to avoid further quality loss. those that use frozen blocks of whole fillets. Fish cake cores may be batter-enrobed. pathogens such as Salmonella and Listeria. Waste fish generated as a by-product of another process will always be susceptible to spoilage and water loss. chilled. material will need to be transferred swiftly to a chiller. spoil more rapidly than the dry ingredients. fish fingers).7 days.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. as the physical structure of the material has been destroyed. off-cuts and by-products from other processes (maybe minced) are reformatted give a final product that is still 100% fish.g.3.4. Shapes may then be cut from the individual slabs using either hydraulic shape cutters or continued cutting with band-saw type blades (e.1. 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. The fish component of the cake will. or those where the trimmings. more seriously. with the coating heat-fixed. with an expected shelf life of 6 . 2.
e. and Enterobacteriaceae from flour. which can occasionally be isolated from protein-handling processes. e. 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. chilled. 2.g. IQF/pre-formed fillets c. starch. such as Listeria and Vibrio spp. quickfrozen. absorb any excess moisture on the surface of the fish portion. rusk or crumb.. Scampi tails are particularly susceptible to contamination with Enterobacteriaceae. spices. the crumb is poured onto the product followed by repeat enrobing and crumbing to complete the five-stage process. egg. from flour. and if required. potentially. is the addition of a pre-dust. perfringens could. some of this initial flora 36 . which typically consists of flour. or sold as a chilled product after thawing. The first stage. Natural fillets d. Breaded or battered fish fillets are fully heat-processed before consumption. with the coating heat-fixed.or fivestage) frozen or chilled. milk powder and spices. If a crumb coating is required. any excess is removed with an air knife. flour. and to carry flavourings where required. as well as ensuring that there is minimal contamination from human processors. Tempered band-sawn materials b. Fish products. because of the intensive handling required during processing. Products are preformed and presented to the process (either three. A large variety of product formats and types can be batter-enrobed. Spore-forming anaerobes such as C.FISH AND SEAFOOD In all of the above examples. rapid automated processing with the minimum of delays will minimise product spoilage by pseudomonads.4. Bacillus spp. raising agents.2 illustrates the typical stages of enrobing or breading. rapidly multiply if strict temperature control was not maintained. with an expected shelf life of 6 . Initially.4 Battering/enrobing process Figure 2. The function of this is to improve the adhesion of subsequent coatings.7 days. fish cakes Portions or fillets may be batter-enrobed. a characteristic flora will develop from the batter components. and possibly Staphylococcus spp. e. i. a. Control of temperature limits the potential for growth of spoilage organisms and pathogenic bacteria. The product then passes along a wire mesh belt through a ‘waterfall’ enrober that gives an even coating of batter to the entire product.g. and may include milk powder. which is product-dependent and not always required. Batter contains water. Over time. vibrios and other fishassociated spoilage organisms. lactobacilli from the milk powder and flour.
It is advisable that some control mechanism is employed to prevent toxinforming organisms from multiplying to dangerous levels. but it is believed that poor hygiene regarding product and food handlers. The origin of contamination for the two organisms is often unclear. This can be achieved by frequent clean-downs and adequate hygiene. Staph. dry batter mixes are all potential sources. and both of these can produce heat-stable toxins that can survive a batter heat-fix process. cereus can proliferate under certain conditions.and five-stage enrobing/breading process The length of production run and temperature of the batter are key factors in deciding the balance of the dominant flora within the enrober. but because batter is heat-set. contaminants such as Enterobacteriacae can proliferate and develop during the process.2.CHILLED AND FROZEN PREPARED FISH PRODUCTS associated with the ingredients will be diluted out. aureus and B. Flow diagram outline of a typical three. cereus. The FDA has issued clear guidelines on acceptable 37 . this is not of great significance to the microbiological standard of the finished product. Fig.2. in the case of B. time-limitation or production run temperature control. replaced in part by process contaminants and bacterial flora from seafood. and. With inadequate temperature control of batter systems.
There are two main categories.3. pasta or potato. The vegetables must withstand processing. to produce a specific product. Figure 2. The protein source can be fish fillet or block. and must not show evidence of disease. canned fish may be used in their preparation owing to it having wider availability and low wastage. as this could introduce off-flavours and reduce shelf life.5 Recipe products These products involve the combining of seafood with other food ingredients as part of a recipe. 2.1 ºC for no more than 3 hours is recommended (1). rice. A temperature no greater than 10 ºC for 12 hours cumulatively or 21.5. For example. Vegetable quality is an important characteristic often overlooked in the selection of vegetables for frozen and chilled products. and may include molluscs or prawns that may or may not have undergone a minimal heat process during their primary processing.FISH AND SEAFOOD temperature regimes and run times permitted for batter enrobers.3 illustrates vegetable processing and treatment for inclusion in chilled and frozen fish recipe dishes. but they are supplied without a source of carbohydrate e.2.4. Well-managed breading and pre-mix systems that are segregated from batter enrobers present little hazard (7).g.1 Seafood recipe meal centres (frozen or chilled) These products can be defined as the central component of a meal.4. Flow diagram outline for recipe dish ingredients preparation 2. Fig. 38 .
Any assumptions regarding the application of good agricultural practice to produce should be made with caution.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.3 log reduction in bacterial numbers on the surface of vegetables. colour and water will occur. The presentation of vegetables has a significant effect on the product quality.5. Washing with a biocide will at best achieve a 2 . chopping and mixing of material. all-year-round fresh vegetables are preferred. The majority are products made from various types of whitefish (cod. Blanching of vegetables can be an effective means of reducing or removing surface contamination.e. i. Salmonella typhi. This is a concern for chilled products that may have a shelf presence for up to 6 days including day of manufacture. the use of night soil or incorrectly applied manure or sewage sludge for ground preparation may contaminate produce with a wide range of pathogens. coli O157. and reduce spoilage organisms and potential pathogens. In an attempt to improve washing and reduce chlorine taint. with the accompanying release of additional nutrients by processing. with cases of foodborne disease traced to Listeria monocytogenes. The opportunity for growth of L. spore-formers. Products must be washed thoroughly to remove pests and soil. For example. larger diced pieces allow for easier handling and a reduction in spoilage but are less attractive to the consumer. and protected from any post-processing contamination.4. monocytogenes should be taken into account when setting shelf life or holding times of vegetable components. For example. haddock. acidification of hypochlorite and chlorine dioxide have been proposed and recommended as an improved alternative. However. treated as a highrisk ingredient.CHILLED AND FROZEN PREPARED FISH PRODUCTS The products will often be presented with vegetable toppings. coliforms and pseudomonads throughout the mix. not just as accompaniments. they should be stored under temperature-controlled conditions. will disperse the saprophytic flora of vegetables . (7. Obvious signs of vegetable soft rot due to mechanical damage or infection by Erwinia spp. loss of quality. For premium products. and mixed whitefish). even those with pH-modified dressings. encourage more rapid decay and loss of structural integrity. Raw material is typically fish fillet. and E. for example. frozen block or canned fish. Although the processes are claimed to be superior to using unmodified hypochlorite for washing.lactobacilli. or breadcrumbs. control is more onerous. 39 . 2. coryneforms. and may include molluscs or prawns that may or may not have undergone a minimal heat process. 8) will. A number of examples of direct faecal contamination of vegetables exist in the literature (8). if the process is not appropriately controlled. Once vegetables have been blanched. any processing stage. and may include sauce accompaniments. pastry. rice or pasta. but also as components in recipe meals. However.
cereus). B. Typically. if subjected to temperatures between 20 and 40°C. cereus. such as butter garnish with black pepper. 2. For this reason all products. a process equivalent to at least a 5D Listeria kill will be necessary.4. Placing such products in a standard chill store to cool down could result in condensation. Appropriate handling facilities or equipment will be required to protect the product. monocytogenes and other bacteria that may be present on chilled surfaces such as ceilings. such as high-risk/low-risk barriers and dispensing and/or holding at above 63 ºC. the preparation process must ensure the eradication of all vegetative pathogens from the ingredients. On hydration. Products or dried ingredients contaminated with either spores of aerobic bacteria (e. 11). If the sauce contains modified starches for freeze/chill stability.6 Preparation of non-fish components of chilled and frozen recipe dishes 2.g. a heat process in excess of 73 ºC will be required to rehydrate this component fully. Potato toppings made from flake or powder will normally have little associated spoilage flora. with the range always expanding as new tastes and cuisines are developed. and an increased risk of contamination from L.) to the product (12.6. but temperature-abused pasta can also spoil rapidly by mould growth. rely on the use of microbiologically sound ingredients. or use barrier or hurdle principles to preclude the growth of pathogenic microorganisms. and then cooled slowly allowing germination and re-growth. especially in those products that have a vegetable component or rice accompaniments.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. aureus toxin is well documented (10. Spices and herbs added to form toppings along with breadcrumbs or other coating materials will similarly contribute a spore loading (primarily Bacillus spp. This is a particular issue especially where products are heat treated to eliminate vegetative cells. should be chilled to <5 °C as quickly as possible using either blast chillers or other equipment specifically designed for this purpose. Manufacturers of chilled products with an extended shelf life must recognise the potential for growth and spoilage of psychrotropic strains of B. 40 . spore-formers can proliferate if adequate control of temperature is not maintained. cereus are well known and are still a common issue in the UK (9).FISH AND SEAFOOD Food-poisoning incidents traced to rice and caused by B. Chilled pasta is becoming more popular than dried pasta as a component in many product lines. 13).4. Alternatively. can germinate and rapidly multiply. An incident involving Staph. where composite meals are heat-treated. Sauces or marinades are added to fish to broaden appeal or to accompany components.
for instance. Without barriers. spices. etc. Figures 2. 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. suspended in an acidified sauce with an oil component.2.5 illustrate the recipe dish ingredients preparation and assembly and packaging. flour products. respectively.4.4 and 2. 41 . 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. 13). 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. that of a dairy-based sauce. Marinade-type products are becoming more popular.CHILLED AND FROZEN PREPARED FISH PRODUCTS Fig. Acidified or low-pH sauce formats using acetic acid or vinegar normally require a lesser heat process for the equivalent effect. and handling of processed sauce can be less rigorous than. Flow diagram outline for recipe dish ingredients preparation Time limitation may minimise the potential for spoilage by process contaminants or at the dispensing unit.herbs. The format uses flavour compounds .
the method replaces air in the sealed packaging with a defined mixture of gases.2. generally carbon dioxide and nitrogen.5. which may restrict either microbial growth or enzyme activity.7. The objective of this method is to deny the aerobic bacteria (both spoilage and pathogens) the oxygen that they need to grow. These include: 2. 42 . unless the temperature is strictly controlled to below 5 °C throughout the supply chain. Flow diagram outline assembly and packaging of recipe dish 2. and tuna loins are packed by placing the product in a plastic pouch and removing all the air. botulinum to grow.4.2 Modified-atmosphere packaging Originally developed for the red meat industry. Increased levels of carbon dioxide will also result in a slight decrease in the pH of the surface tissues. as it dissolves in the surface moisture to form a weak acid. so replacing the oxygen necessary for aerobic growth of spoilage microorganisms. 2.1 Vacuum packing A range of fresh chilled products including salmon (smoked and fresh portions).FISH AND SEAFOOD Fig.4.7.7 Effects of packaging on chilled products Products with minimal additional ingredients may use packing techniques designed to minimise microbial growth. The oxygen-free conditions will however allow other anaerobic bacteria such as C.4.
It is not clear whether the additional contaminants from components or processing will accelerate this typical seafood-type spoilage. This is considered to be the one major difference between fish and meat products. This is also true of chill storage and the distribution chain. Aeromonas spp. The case for growth and contamination of products with Aeromonas is stronger. and therefore will have a greater opportunity to dominate other spoilage flora numerically over the course of product shelf life (15). at slightly higher temperatures (10 ºC). Spoilage will be most readily detected in proteinaceous components of ready meals. Beyond this. by a loss of texture. phosphoreum. initially. but they will also be readily adapted to the nutrient source and conditions. the slow development of ammonical and sulphuric ‘off’ odours and flavours(16). and Yersinia spp. have both been suggested as possible spoilage or pathogenic contaminants of chilled recipe meals (18). At refrigeration temperatures (5 . Not only will these organisms have a numerical advantage over other proposed spoilage organisms. but little on in-pack spoilage.1 Chilled storage For chilled products or chilled components of frozen products. 43 . Vibrionaceae will succeed (17 ). Nonetheless. is typically uncooked or at best lightly processed to retain eating characteristics.CHILLED AND FROZEN PREPARED FISH PRODUCTS 2.5 Spoilage During processing. in numerical terms. Shewanella putrefaciens and Ph. both organisms are capable of growth at chill temperatures.10 ºC) Enterobacteriaceae are often found to proliferate and produce a characteristic type of spoilage that includes. but a conclusive case for pathogenicity has yet to be presented for Aeromonas spp. (20). The fish component of recipe dishes. Spoilage of this material is not expected to differ greatly from the processes described elsewhere. a large number of nutritionally non-fastidious spoilage organisms are commonly isolated from seafood.5. as this organism is a common contaminant of freshwater fish species (19). but is not limited to. where the protein component is nearly always heat-processed. and finally flavour loss. 2. followed by moisture loss. The literature has good information on spoilage of individual components of recipe meals. advanced spoilage gives further changes in texture and the production of characteristic ‘off-odours’. and in influencing the final eating quality of the product. 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.. except for some crustacea. However.
botulinum. Characterisation of the enzymes has demonstrated residual activity even after a heat process that would destroy vegetative cells (24). However. released by a pseudomonad with demonstrable activity to 0 ºC and a peak in production at 10 ºC (15). Bacillus spp.g. Serratia spp. freezing also has a protective role in improving the safety of raw materials by. also be those that have the least effect on the viability of microorganisms (21). Spoilage can be either a function of bacterial growth or the microbial release of free enzymes into solution. As a note of interest. auto-oxidation of fats/oils). and Aerobacter spp. Pseudomonas spp. The conditions and freezing process most favourable for good-quality products will. It is not unreasonable to expect that spoilage bacteria release many extracellular enzymes. Products that are developed with an extended shelf life beyond the 6 .3 Enzymic spoilage Post-harvest changes in the quality of fish due to enzymic activity and rigor mortis are well known and understood (10). producing characteristic off-flavours. This also ensures that any post-process bacterial contamination or spore-forming bacteria in the product will not grow.. for example.5. This reinforces the need for prompt processing and temperature control of ingredients and finished products. where the variety of components and ingredients used in manufacture is varied and complex. within dairy products are well known. 2.. however. a recent report examining fish gut bacteria from cold waters characterised a proteolytic enzyme. Still. the shelf life of products is limited by the natural spoilage process described earlier. 44 .5. eliminating nematodes in products that do not have a heat treatment step. spoilage should not be considered a purely microbial phenomenon as taints and off-flavours can develop by non-microbial means (e..7 days typical of fish products/recipe meals must consider the potential for growth of non-proteolytic C. this should also take into account shelf life of product components used in the assembly of the final product.FISH AND SEAFOOD 2. The type and nature of the enzymes and their activity will depend on the bacterial species. For example. By retaining the intrinsic spoilage flora of seafood. 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. Other examples of bacterial proteolytic and lipolytic enzymes produced by Alicaligenes spp.2 Frozen storage Maintenance of product temperature at <-18 ºC will not permit bacterial growth or enzymic spoilage. These have been proved to impart off-flavours. which would warn the consumer of the potentially unsafe nature of the product. enzymic processes are often ignored or overlooked as insignificant or unimportant because of the rapidity of fish spoilage.
2 Clostridium botulinum There are reports of rapid spoilage of vegetables by C. 2. cereus. 45 . agricultural run-off or sewage can contaminate in-shore species. spices and herbs and vegetable shoots have been reported (12. 2. with pathogenic microorganisms or viruses. and this issue may be of greater consequence for warmwater.1 Bacillus cereus This is a common contaminant of dust. histamine intoxication.6 Pathogens: Growth and Survival There are only two species of pathogenic bacteria that can truly be said to occur naturally in fish. and many raw or processed foods (26).CHILLED AND FROZEN PREPARED FISH PRODUCTS 2. Numerous quoted figures exist in the literature regarding cell counts required before toxin production occurs. in some cases. botulinum when held in favourable conditions for growth (28) i. perfringens in seafood noted a transient population of spores in the guts of fish feeding adjacent to rivers and sewage outfalls (29).. Additional hazards from marine bio-toxins. water. low oxygen or anaerobic conditions. Cases quoting contamination of seafood with organisms such as S. air. potato. especially molluscan filter feeders.104/ g should be treated with some concern (26). and some effects related to product type and time to toxin production have been noted. are dealt with elsewhere in this book. Vibrio spp. 27). A number of other Bacillus spp. typhi. Therefore. any risk of C. In addition. or a heat shock followed by holding at a temperature suitable for growth. soil. Hazards attributable to the non-seafood components are listed below and this information. perfringens contaminating seafood will depend on the source of raw material. and Campylobacter are recorded in filter feeding molluscs such as mussels. Counts of between 103 . farmed or free-living crustacea (30).6. permit rapid growth of B. etc. 2. may be of benefit within any risk assessment for chilled and frozen prepared fish products. botulinum type E and Vibrio parahaemolyticus (25). have been linked with food poisoning and should also be considered. Germination of spores requires favourable conditions. Incidents in such diverse materials as cream. These are C.3 Clostridium perfringens A study examining C. along with previous discussion. This may occur where storage under gas such as carbon dioxide is used to reduce ripening and so prolong storage.6.e.6. 13. Inappropriate holding times and temperatures will.
but. and inadequate cooking or holding within the temperature growth range (27). These include inadequate refrigeration and preparing foods far in advance of planned service. 2.6. which is not affected by any subsequent cooking process. L. infected food handlers. at the present time.FISH AND SEAFOOD Vegetable raw materials are potentially contaminated through soil conditioning with animal manure.6. It is unclear. Chilling may help Shigella to survive. aureus (35). aureus enterotoxins in foods. Rates of contamination with smoked seafood are much higher than those with unsmoked products. L. but under the appropriate conditions.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). when introduced or inoculated into sterile components of chilled prepared recipe dishes. if any other potentially pathogenic organisms. All the above points should be considered within any risk assessment of the potential for growth and toxin production by Staph.6. These conditions will allow the production of the toxin. the possibility of a sanitiser-resistant biofilm colonising processing environments should be viewed with some concern (32). Sources of contamination are many because of the widespread distribution of this organism in the environment. monocytogenes as a contaminant in foods (31).6 Staphylococcus aureus There are a number of underlying reasons for the development of Staph. monocytogenes has been frequently isolated from fish-processing plants and is thought to be able to colonise this type of environment (32.5 Shigella spp. An acceptance has now developed in the UK of the presence at low levels of L. 33) due to its ability to grow at low temperatures and form “biofilms” on surfaces. growth may occur in foods (26). 2. indigenous or other. Shigella spp. and this is believed to be due to a tolerance to brine used in the smoking process (32). will colonise factory environments in a similar fashion except for possibly Vibrio spp. 46 . However. and will therefore cause “food poisoning” even in a product that is cooked before consumption. the restrictions on growth rate seen with seafood will not apply. monocytogenes tends to grow poorly at refrigeration temperatures in seafood (33). 2. are considered primarily to be waterborne organisms but epidemiology indicates that contamination of vegetables and possibly in-shore caught seafood could act as vehicles for infection by Shigella (34).
and not to final products. and as such the monitoring is for verification purposes rather than control.6. and are a recognised means of providing data on manufacturing standards.7 Published Microbiological Criteria Due to the complexity of many processed food products.CHILLED AND FROZEN PREPARED FISH PRODUCTS Illnesses caused by Staph. etc. For instance. It should be remembered that 47 . but provide the industry with an indication of the microbial level at which action is required to ensure product safety.III based on IFST Guidelines (39)) provide example guidelines for seafood products. . aureus enterotoxins and B. with respect to the number and types of ingredients used. but these may be eliminated by a subsequent heat-treatment stage.II and 2. The following tables (Table 2. such as batter mixes. 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. (36). as published by for example. it is not surprising that Microbiological Standards (defined as mandatory compliance required (37)) are limited to those raw materials in which specific hazards occur. cereus emetic toxin are often confused owing to similar symptoms. are not mandatory. although due to long analysis times required. pastry. Microbial guidelines. Microbial levels at the end of manufacturing (m) will be set much lower than those at the end of shelf life (M) which in turn should still be safe to consume. pasta. Microbial criteria and the monitoring of microbial levels within a product are important for monitoring trends within production systems. and their association with similar materials. In the EU these are defined under European Commission (EC) Regulation 2073/2005 (38). the fresh product is often consumed before the results are obtained.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. Microbiological specifications do not necessarily relate to product safety. 2. unless the criteria is for a pathogenic organisms where only non detection is acceptable. 2. the Institute of Food Science and Technology (IFST) (39) or the British Retail Consortium (BRC) (40). although the only seafood products specifically identified are raw “shelled and shucked products of cooked crustaceans and molluscan shellfish”.A dairy-based sauce that includes a non-pasteurised cheese will contribute significant aerobic total counts. 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.
III IFST Guidelines for processed foods . batter etc) should also be consulted when establishing the specifications for raw materials and final product. parahaemolyticus (warm-water fish) Histamine (scrombroid fish) Indicators and spoilage organisms Organism APC (heat treated) Enterobacteriaceae E. 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. fish fingers. pasta. ready meals Storage: Frozen/chilled Use: Re-heated prior to consumption Pathogens and toxins Organism Salmonella spp. cereus Staph. fish cakes Storage: Frozen/chilled Use: Re-heated prior to consumption Pathogens and toxins Organism Pathogens GMP* Maximum Criteria for absence not generally applicable Indicators and spoilage organisms Organism E.II IFST Guidelines for part-cooked foods – e. aureus V. TABLE 2. monocytogenes B.FISH AND SEAFOOD the microbial criteria for other products (rice.g.fish products. Maximum= Levels at end of stated shelf life 48 . L. coli GMP* <102 Maximum 104 TABLE 2. coated fish.
663-87. 1997. Woolaway M. 67-73.. 373-8.C. 204-16. Aureli P. 1991. Bartram P.K. 24 (5)..M. Beuchat L.R. Letters in Applied Microbiology..G... 1982.R. 7. 6. Proper processing and handling of raw tuna. Behling A.M. Ishizaki K.A. Recent Advances in Food Science. 25 (1). 15.CHILLED AND FROZEN PREPARED FISH PRODUCTS An authoritative survey of specifications can be found in Shapton & Shapton (41). London. Ltd. 10..W. Lawyer R.. 2.R. .H. 2. Letters in Applied Microbiology. Food Science and Technology Today. 8. Journal of Food Science. Powers E. Taylor S. Microbiology of processed spices. Outbreaks of foodborne illness in humans. 59 (2). Christiansen T. 7 (24). Yumoto I.. Gilbert R. Microbiology of Marine Food Products. Wienke A. IBH Publishing Co. Communicable Disease Report. 167-93. International Commission on Microbiological Specifications for Foods. 1993. Cambridge. Murrel H. Communicable Disease Report. Microorganisms in Foods 6: Microbial Ecology of Food Commodities. 49 4. Huss H. 11.. Journal of Hygiene.. 166-73. 1976. Animals and Spoilage.M. 47. Fish and Fisheries Products: Hazards and Controls Guidance... Ohgiya S. Hobbs G.. 1976. Tu A.. 13.. Dalgaard P. 1962. Journal of Food Protection. 9.L. Martin R. 1. Importance of Photobacterium phosphoreum in relation to spoilage. Powers E.T. Shewan J. Washington D.J. 1997. 96. Kumeta H.. References US Food and Drug Administration. 1997. Toxin Related Diseases: Poisons Originating from Plants.R. in Fish Inspection Quality Control and HACCP: A Global Focus. 2005.. 233-7. Kluwer Academic. England and Wales Quarterly Report. Eds. 668 -70. 3. Collette R. 5 (3). 14. International outbreak of staphylococcal food poisoning caused by contaminated lasagne. Hoshino T. Hackney C. New York.8 1. Pvt. Masuoka Y. 39 (10). Incidence and levels of Bacillus cereus in processed spices.C. 1986. Journal of Milk Food Technology. 5. 215.. Pathogenic microorganisms associated with fresh produce..R. The bacteriology of fresh and spoiling fish and some related chemical changes.C.E.. Isolation of a Pseudomonas species from fish intestine that produces a protease active at low temperature. 1997. 12. 70-2.L. Food and Drug Administration.. Matsuyama H. Latt T. Mejholm O. Ward D. Taylor and Francis. Slavin J. Bacterial histamine production as a function of temperature and time of incubation. Fish: microbiological spoilage and safety. Journal of Milk Food Technology. 38 (11). 2001. 1311-7. 1991. London. Van Nostrand Reinhold. Bartlett C.L. Brown T.. Sakamoto T. Oxford.J.
34-40.R. Journal of Food Safety. 24. 1995.. Blackie Academic Press.FISH AND SEAFOOD 16. 1988.Yersinia. 59-72. 51-158 Saxby M. Food Microbiology. Advisory Committee on the Microbiological Safety of Foods. 25. Detection of specific spoilage bacteria from fish stored at low (0 ºC) and high (20 ºC) temperatures. Anon... Jemmi T. 23. Report on Vacuum Packing and Associated Processes. International Journal of Food Microbiology. 13 (1).A Literature Survey. Leatherhead Food Publications. 26. Survey of the incidence of Aeromonas and Yersinia species in retail foods..C. 28. Jay M. Blackwell Scientific Publications. Gould G. 22. What problems does the food industry have with spore forming pathogens B. Curran D. 1977. in The Microbiological Safety and Quality of Food. Her Majesty’s Stationery Office. . 2005..W. 1994. Matches J. 30. 19. 31. cereus and C. Barnes E. 28 (4). in Anaerobic Bacteria in Habitats Other than Man. Food Research International. Keusch A. London. PHLS Microbiology Digest. 4 (1).M. Lund B.. Glasgow.. 1996. Brooks J. 65. Leatherhead. Granum P. Huss H. Clostridium perfringens in the environment. Liston J. Aspen Publishers. 1974. 351-72. 309-16. 155-9. 4. Gram L. 1986. Eds. 25 (2).C.J. U. Journal of Food Protection. Applied Microbiology. Walker S. 21. Food preservation by combined methods. Microbiological guidelines for some ready-to-eat foods sampled at the point of sale: an expert opinion from the Public Health Laboratory Service.. Fishing News Books Ltd. 1990. coli (VTEC) . 274-86. Aeromonas and Verotoxigenic E. 28... Microbiology in fishery science. Gram L. 41-3. Is Aeromonas spp. 20.H.J. 1992.K. 29. Liston J.J.. a foodborne pathogen? Review of the clinical data..J. International Commission on Microbiological Specifications for Foods.E. Wood L. 50 17. 472-506. Springer. perfringens? International Journal of Food Microbiology.. 1992. Morgan D. Golden D.C..R. 1996. 1979. 1996. The Emerging pathogens . Halligan A. Huss H. 9. Ray B. Leistner L. Modern Food Microbiology. Ed. 13.A. New York. Fresh and processed fish and shellfish . Baird-Parker T. 2000. 27. Andersson A. London.H. Eds.M. Blackie Academic and Professional.J. 18. Microorganisms in Foods 5: Microbiological Specifications of Food Pathogens. Food Taints and Off-flavours. 1987. Connel J.V. Lund B. Oxford. SAB Symposium Series No.. Loessner M.. 333-6. Trolle G. 655-60. Food Control. 32. 11. Effects of freezing and storage on microorganisms in frozen foods: a review. Gaithersburg.L. Anaerobes in relation to foods of plant origin. Occurrence of Listeria monocytogenes in fresh water fish farms and fish smoking plants. Ronner U. Mead G. in Advances in Fish Science and Technology. 40 (5). Speck M. 1993.
pdf Commission Regulation (EC) No 2073/2005 of 15 November 2005 on Microbiological Criteria for Foodstuffs. Smith J. 41. 1991..L. Shigella as a foodborne pathogen. London. Chilled Food Association.eu/LexUriServ/ LexUriServ. 34.co.CHILLED AND FROZEN PREPARED FISH PRODUCTS 33. Journal of Food Protection.H. 2006. U. CRC Press. 50 (9). Gloucestershire.org/Resources/Chilled%20Food%20Association/Public%20 Resources/BRC_CFA_Micro_Criteria_Guidance_Ed_1. Microbiological Criteria. Varnam A.uk/cday/posters2008/npd6. 1998. Campden & Chorleywood Food Research Association. 51 . Cambridge. http://www.G. Phil Voysey. Chilled Food Association. Woodhead Publishing Ltd. Principles and Practices for the Safe Processing of Foods. http://www. London. http://eur-lex. Evans M. 60 (4). 39.campden.pdf Shapton D.do?uri=CELEX:02005R2073-20071227:EN:NOT Institute of Food Science and Technology.chilledfood. 1987. McCarthy S. 1999. 35. British Retail Consortium. Institute of Food Science and Technology.K. 1997.A.F. Kraft A.2.K. 1992.europa.. 36. 37. 40. Shapton N.A. Journal of Food Protection. Psychrotrophic Bacteria in Foods: Disease and Spoilage. Development and Use of Microbiological Criteria for Foods. 788-801. Incidence and survival of Listeria monocytogenes in ready-to-eat seafood products. U. Wolfe Publishing. Foodborne Pathogens an Illustrated Text.A. 38. Guidance on the Practical Implementation of EC Regulations on Microbiological Criteria for Foodstuffs. 372-6.
Pelecypods have little need for locomotion. the adductors. which includes oysters.S. Strong muscles. oysters and clams can process about 3. mussels. MOLLUSCAN SHELLFISH Carlos Abeyta. snails. Other species of shellfish such as crabs. S. the shell. In the process of filtering water. The class having species of economic and public health importance is the Pelecypoda. and shrimp. Box 3012 Bothell WA 98041-4421 United States of America 3. cockles. viruses. Bivalves feed by filtering large volumes of water across their gills to obtain oxygen and food. P. Many microorganisms ingested by shellfish survive the digestive process (3. mussels. squid.1.7 litres of water per hour. slugs. lying well within the mantle cavity. In most species. The digestion process requires less than 2 hours in actively feeding adult bivalves (2). O. Plankton and algae form the bulk of their diet from seawater. abalones.1 3. where it is eventually swallowed and digested. composed of two valves hinged together dorsally. completely encloses the body. Special tracts of cilia move this mucus toward the mouth. Pelecypoda. clams. Food and Drug Administration 22201 23rd Dr. chemical contaminants and other impurities on the mucus of the gills. and tooth shells are a group of soft-bodied animals that usually secrete external protective shells. lobster. whelks.E. cockles. cockles and scallops. they also trap bacteria. octopi. The term shellfish in this context is limited to oysters. mussels. 4). and scallops. the foot is compressed to form a muscular spade for digging and the head is greatly reduced. Pacific Regional Laboratory Northwest U. In these forms. clams. Jr.1 Definitions General features of the mollusc The phylum Mollusca. can hold the shells tightly shut against enemies (1). while 53 . The class Pelecypoda includes all of the bivalves such as clams. oysters.3. The phylum consists of five classes: Amphineura.6 litres. respectively. and 2. Gastropoda. and Cephalopoda. For example. Scaphopoda. scallops. chitons.
Mactra sachalinensis Saxidomus giganteus Japan Pacific – North American Saxidomus nuttali Protothaca staminea or Paphia staminea 54 .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 . Clams are benthic and live burrowed in the mud. Whatever contaminants are in the water will eventually get into the shellfish.North America Saxidomus nuttali Venus mortoni Protothaca thaca Meretrix spp.1. and usually at higher concentrations than found in their habitat.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. many species of bivalves are edible (Table 3. dwelling on the bottom or attached to structures in the water column.2 International commercial products Globally. 3. The habitat of oysters and mussels are sessile. The following are descriptions of the various bivalves and their forms of preparation for human consumption (6): TABLE 3. are not as vulnerable to contamination through pollution of their beds as the bivalves. Mercenaria mercenaria Venus mercenaria Mya arenaria Country Scientific nomenclature Hard clam (also used for Quahaug/quahog) Pacific .I).FISH AND SEAFOOD commercially valuable. but maintain contact with the water by means of a siphon tube (5).
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.
hard-shell or soft-shell clam. A mantle tissue encloses the body of the clam. Buried clams leave tubes.1. but 56 . laterally compressed. The muscular foot is used to burrow in mud or sand. Different species are variously called quahog. or siphons. cherrystone. Jacques Iceland scallop Bay scallop Weathervane scallop (also called Alaska scallop) Atlantic North East Atlantic North Atlantic West Atlantic Pacific – North America Pacific – North America Pacific – Japan Common scallop Sea scallop (also called giant smooth) Japan Atlantic – North America Pecten varius Pecten maximus Chlamys islandica Argopecten irradians Pecten caurinus Pecten aequisulcatus Pecten laquaetus Pecten yessoensis Penten magellanicus or Placopecten magellanicus Pecten meridionalis Pecten novaezealandiae Pecten jacobaeus Aequipecten gibbus Chlamys opercularis Chlamys varius Amusium balloti Mediterranean – South East Europe New Zealand Australia Mytilus galloprovincialis Mytilus canaliculus Mytilus planulatus Commercial scallop Australia New Zealand Great scallop Calico scallop Queen scallop Variegated scallop Scallop saucer Atlantic/Mediterranean Atlantic – North American Atlantic Atlantic/Mediterranean Australia 3. Most clams are only a few centimetres in their maximum dimensions.1 Clam Clam is the common name of a large group of often edible. littleneck. and two symmetrical shells (valves) held closed by two large muscles and joined by a dorsal hinge joint.2. extended above the surface to maintain water currents needed for respiration and feeding. geoduck.FISH AND SEAFOOD Mussel Mussel Mussel Scallops Scallop Coquille St. mostly marine bivalve molluscs. and other names.
in cans with either brine or clam liquor. uncooked meats packed unprocessed. then heat-processed. skewered with bamboo sticks. Frozen: in shell. Canned: clams are steamed open and the meats removed from the shells. tidal waters or in brackish water in the mouths of rivers. Oysters lack a foot or have a reduced foot. 3.1. all in cans.2. Frozen: shelled meats. uncooked. Dried: shelled meats. packed in weak brine. Dried: Hoshigai (Japan) shelled meat. These molluscs feed on minute plants and animals carried to them by the current. boiled and then sun dried for 3 to 10 days before packing in boxes (Hong-Kong) Smoked: meats usually canned in edible oil. uncooked (shucked). oyster soup. oyster bisque. Live: in shells. They live free on the bottom or adhere to stones or other objects in shallow water along the seacoasts. the meats are smoked and packed in oil in cans or jars. either whole or minced.2 Oyster Oysters are marine bivalve molluscs. having rough and irregularly shaped shells that are closed by a single adductor muscle. Smoked: meats. smoked meats are also canned. 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. Fresh: in shell. and no siphon. washed. trimmed and packed. after precooking and brining. Soups: oyster stew. 57 . or shelled meats. hermetically sealed. Canned: meats removed from the shell by steaming. Live: in shell Fresh: in shell or shelled meats. or shelled meats. clam madrilène and clam liquor. sun-dried. Canned soups include clam chowder.
They have a radially ribbed shell with the edge undulated and swim by opening and closing the valves.1. also with other ingredients. or packed with jelly. sauces. or filaments. in mussel liquor. Semi-preserves: cooked meats with vinegar-acidified brine and spices. and the anterior end smaller and pointed. mussel in butter sauce.4 Scallop Scallops are marine bivalve molluscs living in shallow waters of protected bays with sandy or muddy bottoms. Fresh: shelled meats (often only the adductor muscle and the roe are eaten. In some species. boiled. Canned: cooked meats.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. The shells are fluted and round with a broad. Unlike other sessile bivalve molluscs. Fresh: cooked meats. in which the individual branches.) Dried: peeled. scallops possess a single large adductor muscle. smouldered and afterwards dried (Japan).2. The familiar marine mussel is asymmetrical. 58 . mussel paste. The adductor muscle is the only edible part of the animal. composed of striated muscle fibres (used for fast swimming) and unstriated muscle fibres (used to hold together the two halves of the valves). Smoked: meats may be canned in edible oil. cooked meats packed in dry salt for transport.FISH AND SEAFOOD 3. the shell is often characterised with a lustrous nacreous or pearly lining inside their shells. Bait: used live extensively for baiting fish hooks for line fishing. are united. and held apart. by interlocking tufts of hair-like cilia. scallops can swim or leap by a series of rapid closings and openings of the valves. brine. flattened ear or wing at the hinge area. vinegar solution.2.1. Salted: cooked meats bottled in brine. The common scallop is about 5 cm long. gutted. with the posterior ends of the shells (valves) broad and rounded. 3. Live: whole with shells. Internally. They have filibranch gills.
also bottled in brine.2 Initial Microflora The microflora of molluscan shellfish directly reflects the environment from which the shellfish is harvested. Because molluscan shellfish are filter feeders. salinity. Salted: meats either lightly or heavily dry salted. Cockles range from about 1 to 15 cm in length and have two valves of equal size.2. clams and mussels. are vectors of disease transmission (8).5 Cockle A marine bivalve mollusc having a shell with convex radially ribbed valves. and the environment. Fresh: meats removed from the shell by boiling. nutrient concentration and pollutants of the water column. 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. they accumulate pathogenic microorganisms from polluted waters. 3. and butter. Live: in shell. 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. which affects the temperature. Primarily inhabits shallow waters near tidal zones where they bury themselves in the sandy or muddy bottoms.MOLLUSCAN SHELLFISH Frozen: shelled meats. in sauce. pH. Canned: in own juice. Whatever contaminants are in the water will eventually get into the shellfish (7). 3. Canned: in brine. mostly in oysters. 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 .1. The microflora are dependent upon a number of factors such as season. Raw and partially cooked shellfish. Vinegar cured: bottled in malt vinegar after brining.
Flow diagrams of the major processing steps of molluscs are presented in Figures 3..3. 12.1 Processing and its Effects on the Microflora Oysters. Enterococci spp. The largest groups found are the Pseudomonads. Micrococcus spp. At harvest. For example. There are microorganisms. 13). The microflora of molluscan shellfish at harvest consists predominantly of Gramnegative rods while the Gram-positives constitute a minor portion of the flora. salinity. They are termed viable non-culturable microorganisms. clams. and yeast cells of Rhodotorula rubra and Trichosporon spp. rakes. Campylobacter jejuni Escherichia coli Hepatitis A Norwalk virus Other viral agents A variety of commensal microflora are found in molluscan shellfish. 11. (9.FISH AND SEAFOOD Aeromonas hydrophila group Staphylococcus aureus Shigella spp. shellstock (live) which is shellfish in the shell. Alcaligenes spp. molluscan shellfish have an Aerobic Plate Count (APC) of approximately 1000 to 100. and Aeromonads. dredges. and shucked shellfish that is shellfish. non-B Snow Mountain virus Plesiomonas shigelloides Salmonella spp.2. whole or in part.. and cockles The processing of bivalves takes two forms.. Acinetobacter spp. 3. spirochetes of the genus Cristispira and Saprospira..1 and 3.1 Shellstock 3. Flavobacterium spp.1 Processing description The method of processing shellstock begins with the harvester collecting shellstock by the utilisation of tongs. Moraxella spp. Water temperatures affect the concentrations of microflora in the water. have been observed microscopically in the digestive tract of Eastern and Pacific oysters. Bacillus spp. 15). mussels. Vibrios.. 10. the initial flora composition and concentration of bivalves are highly variable.1. Campylobacter coli Bacillus cereus Hepatitis non-A. 3. nutrients. from which one or both shells have been removed. observed microscopically.1. or by 60 . but have never been cultured (14.3...1. that have never been cultured on laboratory media.3 3. Other microorganisms found in lesser numbers are the Achromobacter spp.. Temperature..3. pollution point sources and hydrological events will greatly influence the initial microflora in bivalves. In summary. responding to the environment in which they are harvested. Pseudomonas spp.000 /g. mechanical harvesters.
Shellstock are placed in drycooler storage containers in the facility.1. 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. or from other sources approved by the State Shellfish Control Agency (SSCA).1).3. Generic flow diagram of live shellfish from harvester to point of sale 61 . Water used for shellstock washing is from approved growing areas. 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. If by boat. for transportation to the point of sale. after harvesting.MOLLUSCAN SHELLFISH hand (Figure 3. wet storage may be used to increase palatability by increasing salt content of shellfish harvested from low salinity waters. The shellstock is transported dry. in containers from which they can be retrieved easily. or in bay-approved shellfish-growing waters prior to sale or processing. or immediately placed in refrigerated trucks. Thirdly. There are three primary reasons for wet storage: first. Fig. shellfish can be harvested from remote areas at convenient times and held live for brief periods at locations close to the point of sale. Secondly. usually in open-mesh containers and/or burlap sacks. by truck or boat.
At this temperature the multiplication of faecal coliforms (E. Shellstock should be refrigerated at temperatures at or below 10 ºC (50 ºF). hydrophila (18). 23. without adequate refrigeration. The safety of shellfish is predicated by the cleanliness of the growing area waters from which they are harvested. Furthermore. Generally. Several studies have established that pathogenic Vibrios and other bacteria may be present in marine sediments throughout the year (21. Klebsiella. however. Food and Drug Administration (FDA) recommends that potentially hazardous food such as shellfish be held at 7. The washing of sediment and detritus soon after harvesting will prevent further contamination. 24. Wet storage operations are highly variable and may range from temporary storage near-shore in approved areas to on-shore tanks using re-circulating. coli. and the sanitary practices applied during harvesting and shipping. handling. for a considerable length of time. Several studies have established the increase of bacterial counts in shellstock during post harvest period before the shucking process (18. The most important factor in controlling bacterial growth in shellstock is temperature. 17). metabolic waste accumulates in the shell serving as nutrients for bacterial growth. Excessive sediment on the shells and dead shellfish may increase bacterial 62 .2 Effects of microflora during processing The initial microflora found on or in shellstock is influenced by sources of shellstock. Oysters will remain alive in the shell for approximately two weeks if kept at the recommended temperatures. 21. Enterobacter. An outbreak of infectious hepatitis in Sweden in 1956. Harvest conditions are another factor that influences the growth of bacteria in shellstock. Shellfish may contain higher levels of pathogens than are found in the water in which they grow. synthetic seawater for the purpose of de-sanding and salt uptake.1. involving 691 cases was attributed to oysters contaminated in a wet storage area (27). 16. and Citrobacter species) and Vibrios. 26).1. there is evidence that in the absence of adequate refrigeration. some pathogenic organisms will survive and multiply in shellfish (20. 25).FISH AND SEAFOOD 3. Generally. are prevented. vulnificus. with prolonged storage the microbiological quality of the bivalve decreases. 22). the bacterial counts in the meats will increase. including V. If long delays occur before reaching the processing facility and/or refrigeration.2 ºC (45 ºF) or below (19). Washing shellstock helps prevent quantities of sediment and bacteria being mixed with. Shellfish in wet storage tanks are similarly subjected to pollution if the tank water is obtained from a polluted source. shellstock are not refrigerated on the harvest boat. bivalves tightly close their shells thus trapping water and microflora. and thus contaminating the shucked meat at the point of sale. when it reaches the processing facility. There is a positive relationship between sewage-polluted shellfish and enteric disease transmitted by shellfish (8. with the exception of A. Water used for shellstock washing should be of good sanitary quality. Over a period.3. At harvest. and temperature storage of shellstock to point of sale. shellstock are placed in refrigerated trucks for transport or placed in wet storage. to avoid possible contamination of the shellstock.
2. To remove the shellfish attached to the remaining valve. and packing shellfish into containers (Figure 3. washing.2). 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. and fish. perforated tray in which shucked shellfish are spray washed and/or drained) or in a blower-washer (a tank-like device for immersion washing of shucked shellfish).2. Washing of meats with potable water is accomplished by placing the meats on a skimmer table (the stand-supported. In addition. Meats are collected in either perforated or non-perforated buckets for delivery to the washing station. the finished product may become contaminated. Shucking and washing of meats under sanitary conditions prevents the proliferation of bacterial populations. After washing of meats.3. for the effects of microflora during the processing of shellstock.1 Processing description The processing of shucked shellfish is limited to washing of shellstock. a considerable quantity of the adhering material will be mixed into the shellfish during the shucking process. 3. the tank is opened through a drain gate and chute. are applied to the shucking operation of shellfish.2 Effects of microflora during processing The same considerations as previously discussed.2 Shucked shellfish 3. shellstock is subjected to contamination when stored on the floor in standing water. Thus. lobsters. In the blower-washer. removal of shellfish (shucking). washing and culling shellfish prior to storage is essential. Due to the nature of the shucking operation. Studies have shown the use of rubber gloves reduces the contamination level. 3. the knife is used to cut the remaining adductor thus freeing the shellfish (meats). If shellfish are not reasonably clean at the time of processing.184.108.40.206. Shuckers routinely use either cotton or rubber gloves during the shucking operation. through which the washed meats are discharged for rinsing and then packed in containers. It presents a risk of cross-contamination from the non-molluscan animals to the bivalves.MOLLUSCAN SHELLFISH loads in the tanks and lead to increased microbial levels in the shellfish during storage. clothing becomes very soiled. or subjected to splash from foot traffic. Under these conditions if the shuckers enter the packing room. Sealed containers are placed in boxes and covered with crushed ice for chilling or refrigeration and transportation. Another potential risk with the use of wet storage is the commingling of bivalve molluscs with other marine species such as crab. thus contributing to high bacterial counts in the final product (28). whereas cotton become wet and 63 . Containers are placed in packing boxes for shipping at refrigerated temperatures.1. the meats are in a tank of water and agitated by the introduction of air from the bottom of the tank.
2. 3. 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. 64 .FISH AND SEAFOOD retains bacteria during the shucking thus increasing the bacterial load in shellfish (29).2 ºC or less. cleaned and delivered to the packing room or prechilled and placed in temporary refrigeration (≤ 2 hours) at 7. Shucked meats are thoroughly drained. Shucking buckets are completely emptied at the packing room and no overage is returned to the shucker. Separate rooms or lockers for storage of clothing. 2. Shucking containers are rinsed clean with running water. and gloves help prevent the contamination of shellfish. within one hour. 4. Shucking buckets and storage containers are kept at adequate height above the floor to prevent contamination from floor splash. and sanitised before each filling. There is a tendency to store such articles on the shucking benches or in packing rooms. which may lead to contamination of product. Fig 3. aprons.
particularly shucked meats. The initial flora found in shellfish is the major factor of spoilage in bivalves. Achromobacter and Flavobacterium levels decrease during storage. keeping shellfish refrigerated is the most effective method of retarding deterioration and spoilage. Shucked shellfish are held and transported at temperatures of 7. and packing the shellfish in a timely manner. spoilage will occur. 35. Shucked shellfish are an excellent medium for the growth of bacteria. abundant gas production.8% (37). 31. showed that organisms such as Achromobacter. For example. Refrigeration and shipping of shucked shellfish in wet ice is highly recommended. particularly oysters stored at 7 ºC. and yeasts. in oysters. are an excellent medium for growth of bacteria.MOLLUSCAN SHELLFISH Unacceptable practices in the packing step can affect the microflora of shellfish. Temperature abuse will result in high bacterial counts. In summary. Micrococcus. These organisms may play a significant role in the spoilage processes. All equipment coming in contact with the shellfish such as colanders. Much of the work has been done with oysters and clams. As described previously. Glycogen is hydrolysed by the initial flora found in shellfish. Alcaligenes. shucking pails.47 to 6. where there is a breakdown of temperature control or prolonged storage of shellfish. at the turn of the century. 3. is stored in the bivalve as a source of energy. 65 . Pseudomonas. 34. clean gloves. 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. whereas bacterium such as Pseudomonas. 32). Studies have shown that bacterial growth is significantly reduced at storage temperatures of less than 7. 36).2 ºC. Proteus. Shellfish. and proteolysis (10). and that storage in wet ice is the most effective method for refrigeration of shucked meats (30. Lactobacilli seem to be the predominant microflora in shellfish.4 Spoilage There is a lack of information in the literature regarding spoilage microorganisms in shellfish. glycogen levels range from 0. Earlier studies. 33. Flavobacterium. anywhere during the process of shellfish from harvesting to the point of sale. Controls that should be applied include using clean packing equipment. a carbohydrate. thus producing acid and gas (38). these include lactobacilli. Glycogen. The environment has an effect on the bacterial population found in shellfish. streptococci. skimmers. This process changes the bacterial profile to microorganisms favouring low pH. or inaccessible surfaces are apt to harbour accumulations of organic material in which bacteria may grow. and Pseudomonas fluorescens are the responsible agents of spoilage in oysters held at various temperatures (10. The spoilage process in shellfish may be divided into three stages: increase of acidity. blowers and other utensils that have cracked. rough.2 ºC or less.
48 hours with abdominal pain. and harvesting of shellfish in the cooler months. The ability of the bacterium to cause gastroenteritis is highly correlated with the production of a heat-stable haemolysin (42). parahaemolyticus multiplies rapidly at temperatures above 20 ºC.5 Pathogens: Growth and Survival This section deals with bacterial pathogens frequently present in shellfish. The majority of the illnesses were associated with the consumption of raw or partially cooked oysters and clams.5. parahaemolyticus causes gastroenteritis lasting 24 . Sporadic cases are frequent along the coast of the United States. This bacterium is present in all the major processing steps of a shellfish plant. One of 250 cases reported involved shucked products with low levels. ranging from epidemic cholera to sporadic cases of diarrhoea. 44).000 population/year (46). Fortunately. parahaemolyticus possibly producing illnesses (40). 66 . nausea. Between May and September 1997. and estimates based from a hospital study show less than 0. V. parahaemolyticus from shellfish is low. there were over 250 cases of illness on the West Coast of North American extending from California to Canada. shellstock and packing of shellfish in jars) (29). Products involved were from the same processor. 41). there is a recent report of low levels of V. however. lot and retail outlet. Major outbreaks are associated with the warmer months. Descriptions of outbreaks or reports of growth of the organism in shellfish are presented. It is a common marine bacterium. and to the point of sale. The control of growth and survival of these organisms can be reduced by the use of good manufacturing practices including immediate and proper refrigeration. headache and fever. parahaemolyticus. and very common in Japan. although non-hemolytic strains have been associated with illness (43. during processing. They can be introduced to the shellfish from the environment or added during processing. widespread in both polluted and unpolluted waters. Diseases caused by Vibrios are enteric in nature. parahaemolyticus illness is usually a result of eating raw or partial cooked shellfish (8.5 cases/100. These organisms can be present in the shellfish from the point of harvesting.1 Vibrio parahaemolyticus V. 3. The incidence of V. where outbreaks occur regularly. primarily in brackish or saltwater.5. of V. Large numbers of bacteria must be present to reach potentially infective levels of >105 colony forming units per g (39). diarrhoea.e.1 Vibrios Vibrios are Gram-negative facultative rods naturally occurring in the marine water (estuaries. 3. approximately 23 cfu/g.1. from harvesting to finished product (i.FISH AND SEAFOOD 3. only a small percentage of strains in the marine environment are potentially pathogenic (45). coastal areas). prevention of cross-contamination. V.
Onset of symptoms occurs in 48 hours and can be severe. 56). the organism has also been found in cooler temperatures from estuaries of Washington and Oregon (47). and preventing cross contamination once the product is cooked. 23 of 24 major USA-West Coast estuaries were positive for V. High-risk individuals.II (8. these individuals are advised not to eat raw shellfish.1. The bacterium is widespread in estuarine waters particularly in warm waters of >20 °C. vulnificus are fever. 49). Control of oyster-associated 67 . The occurrence of V. however. The widespread distribution of this bacterium is comparable to other studies in the Gulf and Atlantic Coasts (52. cirrhosis. The organism attaches to the intestine and release of a toxin is suspected. chills. primarily in immunocompromised individuals with a mortality rate exceeding 50% (50). vulnificus causing septicaemia and death has been associated with the consumption of oysters (8. cholerae non-01 has been associated with septicaemia. in the range of 0. V.5. Primarily. 57. cholerae. 53.3 Vibrio cholerae non-01 Illness caused by V. leukaemia. fever.2 Vibrio vulnificus The symptoms of V. It causes diarrhoea. 55. Hazards from this bacterium can be controlled by thorough cooking of shellfish. lasting 6 . In one study. 58). cholerae non-01 is ubiquitous in the estuarine environment. 3. 46.0. abdominal cramps.7 days. with a fatality rate of 50% for individuals with septicaemia. V.1. It is one of the most severe foodborne infectious diseases. those who have liver disease. V. Death has been reported to occur within 36 hours from onset. the organism has been found in the Gulf of Mexico. 46. and rapidly become inactive at low storage temperatures. parahaemolyticus the estimated annual incidence rate is very low. Illnesses are usually associated with the consumption of raw oysters as described in Table 3.4 . or immunosuppression. parahaemolyticus growth in shellfish meats is temperature dependent.000 population/year (8. parahaemolyticus. 3.8 case/100.5. cholerae non-01 is a common bacterial cause of molluscan shellfishassociated illness.MOLLUSCAN SHELLFISH Control of V. They are sensitive to cold. All isolates are considered pathogenic. like V. vulnificus. nausea. Oysters are the most hazardous food associated with V. gastroenteritis.48 hours of onset. are particularly susceptible to primary septicaemia. and nausea within 24 . which have been implicated in shellfish-related disease outbreaks (31). cholerae non-01 (51). Control of V. diabetes. Inadequate refrigeration will accelerate the growth of V. cholerae non-01 is generally a less severe gastroenteritis than that caused by V. 54. vulnificus is by rapid refrigeration during warm weather months. or primary septicaemia. Healthy individuals are susceptible to gastroenteritis. 48). The bacterium is an estuarine species that causes wound infections. Again. and vomiting.
including V.2 Campylobacter jejuni C. the pathogenic severity is lower than V. these organisms cause gastroenteritis lasting 24 . however. There is evidence of strains of V. Illnesses are associated with the warmer months. nausea.5%) and 2 . jejuni requires reduced levels of oxygen (3 .5 Other Vibrios spp. parahaemolyticus and V. cholerae non-01 illnesses is by cooking seafood thoroughly. fever. Serotype Ogawa biotype El Tor and 0139 are responsible for epidemics in South American. hollisae. cholerae 01.10% carbon dioxide for optimal growth conditions. cholerae 01 in molluscan shellfish. fluvialis. 68 . mimicus. Illness can include abdominal cramps. It is considered a fragile and sensitive organism to environmental stresses such as oxygen. imported product from a country experiencing an epidemic.5.g. The control of V. It causes diarrhoea. Onset of illness is from 3 to 5 days after ingestion of contaminated food. There are several other Vibrio spp. and shock. and V. and the transmission of the disease by consumption of raw. V. abdominal pain. or cross-contaminated shellfish (57. nausea. In addition. respectively. and southern Asia including India and Bangladesh. dehydration. Gulf coast environment.1. processors should know the product source. undercooked. whereas excellent sanitation facilities in developed countries are responsible for the near eradication of epidemic cholera. In developing countries. 3. vomiting. The presence of these strains appears to be in low numbers and non-virulent (59). acidity and heat. 3. particularly in oysters. In general. vulnificus. is the same as previously discussed for the non-01 strains.48 hours with abdominal pain. salinity.1. The frequency of infection of these organisms is no different from other Vibrios. and nausea. cholerae 01 establishing in the U. V.FISH AND SEAFOOD V. (8. preventing crosscontamination of cooked seafood. 60). widespread in both polluted and unpolluted waters. Poor sanitation and contaminated water supplies will spread the disease. diarrhoea. and harvesting of oysters in colder months when Vibrio counts in water are the lowest (59). 63). Death may occur after severe fluid and electrolyte loss. V.S. drying. They are common marine bacteria.10 days. associated with shellfish-borne illness outbreaks. Control of infection from these organisms is no different from the other Vibrios. 61. 62.5. low pH. and illness lasts 7 . headache and fever. 3. e. Onset of disease is 6 hours to 5 days.5.4 Vibrio cholerae 01 and 0139 Symptoms of epidemic cholera vary from mild watery diarrhoea to acute diarrhoea with characteristic rice water stools. cholerae 01 is transmitted by faecal contamination of food or water.
Campylobacter spp. vulnificus.5. 65. jejuni survived for over 20 days in market-shucked oysters stored at 4 ºC.3. mimicus. V.1 Aeromonas hydrophila group and Plesiomonas shigelloides A. Campylobacter appear to survive well in oysters during storage. V. and V. Recently. V. shigelloides (79. they survive well at refrigerated temperatures. Campylobacters do not survive well in the environment due to the organism's lack of resistance to environmental factors.a (8) Species Hard clams Oysters Soft Clams Mussels Scallops No. 77. parahaemolyticus.500 cells/gram of Campylobacter may cause illness (64. fluviallis. although their role as a pathogen is highly controversial and circumstantial. C. 82. whereas at 10 ºC Campylobacter survive only for 5 days (71). cholerae 01 and non-01. of Incidents 4 279 0 1 0 No. C. jejuni begins in the harvest of shellstock from approved waters. however. In one case. Proper care should be taken if the finished product is to be packaged in modified-atmosphere packaging (MAP). of Cases 4 362 0 1 0 a Includes V. and their immediate proper refrigeration. hollisae Infective dose varies with host susceptibility and virulence of the strain. To reduce the risk of shellfish-borne campylobacteriosis. 70. 78) and P. Human feeding studies suggest that about 400 . 85). 71). 81. 72. They have been implicated in limited shellfishassociated gastroenteritis cases (8.II Molluscan shellfish associated outbreaks caused by Vibrio spp.MOLLUSCAN SHELLFISH TABLE 3. 73). hydrophila group (75. 83. infection occurred among participants of a firemen's banquet. and have been implicated as the causative agent in illnesses from the consumption of raw oysters and clams (68. The seasonality of 69 . 76.3 Other organisms of concern 3. 68. 3. from Pacific shellstock oysters purchased from a retail grocery store (74). This finding is significant in that C. the incidence of Campylobacter infection is low from the consumption of molluscan shellfish. jejuni were recovered. jejuni does not multiply in shellstock at refrigerated temperatures. the organism is being continually inoculated into the environment by waterfowl and farm run-offs and thus eventually contaminating shellfish growing waters.5. 84. 66). ranging from 0. however. Sixteen of 28 participants became ill with campylobacteriosis after ingesting raw clams.4 to 114 cells. 80) are ubiquitous in the marine environment. have been isolated from shellfish beds throughout the world (67. 69. Although. V. control of C.
negative for heat-labile toxin (LT) or heat-stable enterotoxin (ST) 1 Negative for the presence1 Negative for the presence of toxin producing 01 or non-01 organisms 1 <10. non-dairy frozen desserts. FDA/ Administrative Guidelines. FDA/Administrative Guidelines 3 Source (91). prepared sandwiches. after 7 days storage of shucked oysters at a shellfish processors’ cooler kept a 5 ºC Aeromonas counts ranged from 500. 87) of the outgrowth of A. FDA established action levels or levels of concern for certain microbial pathogens in molluscan shellfish. Caution should be exercised by individuals with haematological malignancies (88). The Agency will consider enforcement action against the shipment of molluscan shellfish if the levels exceed as stated. They survive well in shellstock or shucked oyster meats held at refrigeration temperatures. Enforcement action is considered on a case-by-case basis taking into account all the factors associated with the specific situation. 4 FDA and ISSC recommending establishment of an educational program to alert at risk population of presence of V. being isolated more often during the warmer month. action is considered on a case-by-case basis taking into account all the factors associated with the specific situation. They are easily destroyed with mild cooking temperatures. soy products. 2 Source (90). vulnificus in molluscan shellfish. suggesting that these individuals are at higher risk when consuming raw or partially cooked shellfish containing these bacteria. Note: Although molluscan shellfish are not included in the selected high risk food.000 cfu/g1 <230/100 g2 <1000 per gram. cholerae V. aureus C. 3.000/gram (29). and where indicated are intended to be used for official control purposes.6 Published Microbiological Criteria The published microbiological criteria below are included as a guide. vulnificus 1 Acceptable Levels in Molluscan Shellfish <500.000 MPN per gram1 negative for staphylococcal enterotoxin or when the viable MPN count is <10. 70 . dried milk/whey drink powders.600. coli Salmonella V. Pathogen monitoring of Selected High Risk Foods Domestic including: milk. jejuni V.FISH AND SEAFOOD Aeromonas and Plesiomonas is similar to the Vibrios. Control of these organisms in shellfish products is limited. Similar results were seen by other investigators (86. hydrophila group stored at refrigeration temperatures. Test Standard Plate Counts E. coli Enterotoxigenic E.0001 negative for the presence3 Under review4 Source (89). In one study. parahaemolyticus Staph.000 to 3.
Hood M. 1926. 7.. Biology of the oyster in relation to sanitation. Bacterial flora of the market oyster.K. in General Zoology. 33-6. 1994. Hackney C. 1968. 1981. 19-34. W. Microbiological characteristics of Dungeness crab (Cancer magister).M. Occurrence of the spirochaete genus Cristispira in western Canadian marine bivalves. Quantitative and qualitative commensal bacterial flora of Crassostrea virginica in Chesapeake Bay. Cook D. Viability and release Salmonella charity and Escherichia coli from oyster faeces. 1963.. Schmidt C... 26. 8. 1936. Jr. The American Oyster.. 72-8.C. 1970. Ed. Thompson C. A typhoid fever epidemic caused by carrier bootlegging oysters. Washington D. 1964.R.D.C.. Walker W. 13. Effects of harvesting waters and storage conditions on yeast population in shellfish. Saunders Company. Eds. Microbiology of bivalve molluscan shellfish. Lovelace T. 71 . 3.R. American Journal of Public Health. Liston J. Vanderzant C.J. 46. Veliger. Proceedings of the National Shellfisheries Association. 1991. U. American Journal of Public Health. Scanning electron microscopy of Cristispira species in Chesapeake Bay oysters. 82-7. Colwell R. Rippey S. Microbiology of shellfish: bacteriological study of the natural flora of pacific oysters (Crassostrea gigas). Government Printing Office. 1984. Jr. 633-40. Journal of Milk and Food Technology.A. Philadelphia. 30. Infectious diseases associated with molluscan shellfish consumption. 58. New York. 4. Galtsoff P.W.S. Lee J. in Fishery Bulletin 1964 No. Ward D.S. Smith F. Eliot C.7 1. Clinical Microbiology Review. Applied Microbiology. Colwell R. Tubiash H. Journal of the Mississippi Academy of Sciences. water and sediment from Galveston Bay. Van Nostrand Reinhold.. 9. Organisation for Economic Co-operation and Development.S. Tall B. 7. 13. 8... 2. 6. Old H.N.MOLLUSCAN SHELLFISH 3. 6. 15.R. 1975.W. 12. 105-8. 231-51. 1973.S. 1960.E. American Journal of Hygiene. Fate of enteric bacteria in estuarine sediments and oyster faeces. Bernard F.R. Applied Microbiology.. 30. 17. 5.R. 544-8. Nauman R. 71-6.. 33643.L. 755-76.E. Wiley-Blackwell. 1983. 14. Gill S. Journal of Food Protection. 419-25. Fleet G. Eds.. Cook D. Microbial flora and level of Vibrio parahaemolyticus of oysters (Crassostrea virginica). Applied and Environmental Microbiology. 447-52. 42.B. 29. Fish and Wildlife Service. USA. in Multilingual Dictionary of Fish and Fish Products. 36. Galtsoff P.A.F. in Microbiology of Marine Food Products. References The Phylum Mollusca.. Pfeifer D. 10. 1995. 11. 1982. Ray S.R. 104-9. Applied and Environmental Microbiology. Rowse A. 16. Villee C. 64. 1940. 245-7. 44.H.A.K.
72 .. and storage. Maryland.. 343-9. 1983. Takeda Y.. American Society for Microbiology.. Stott R..H. Timetemperature effects on the bacteriological quality of stored shellfish. Fujino T.H.FISH AND SEAFOOD 18. Journal of Food Protection. Chesapeake Science. 20. Public Health Service. Rodrick G. Vasconcelos G. 1980. Olympia oysters (Ostrea lurida). Behaviour of pathogenic bacteria in the oyster.S. Bacteriological studies of commercial shellfish operations on the Gulf Coast. U. I.A. 1992.F. Crassostrea virginica and Mercenaria campechiensis.. 123-8. Part II Harvesting. Ltd. 25..M. Special Report U. Handling and Shipping Shellfish.. Ed. 45..T. Cook D. 1954. 994-1002. Bacteriological changes in live shellfish: Pacific oysters (Crassostrea gigas)..W. Koburger J. Beck W. 1995...E. 167-71.A.C. Influence of U.E.C. Kaysner C. in National Shellfish Sanitation Program Manual of Operations. 1490-5. Ericksen T. 53. Jones W. 19. oysters.A. crabs and mullet. Indicator bacteria and Vibrionaceae multiplication in postharvest shellstock oysters..S. Sadaguchi G. Washington D.G. U.. Presnell M. 23. 1971.S. 4. 1214. Public Health Report. Hoff J. Kelly C. 1205-10. Survival of enteric organisms in shellfish. Colburn K. Ruple A. Temporal occurrence of Vibrio species and Aeromonas hydrophila in estuarine sediments. Hepatitis epidemic conveyed by oysters. Trost P. 69. Liston J. F61-9. Kelly C. re-laying. 1985. Public Health Service Technical Report.. Applied and Environmental Microbiology.W. 1221-8. Applied and Environmental Microbiology. Bacteriological survey of an oyster bed in Tangier Sound.W. 1989. Williams L. Crassostrea commercialis. Ness G. Presnell M. Roos B. Fraiser M. and Manillal clams (Venerupis japonica). LaRock P.B. Saikon Publishing Co. The influence of time and temperature on the bacteriological quality of shell oysters during processing and shipping. 1973.. Son N. 50. 52. 343-5. 30. Abeyta C. Tokyo. 47 (5).W. Hood M..S.. 24.B.S. Eds. Native Littleneck clams (Protothaca staminea).A. 28.. 40. Journal of Food Science.J. 1961. 21.B. in International Symposium Vibrio parahaemolyticus.. 31. Establishment of critical control points in west coast oyster processing plants. 29.J. 324. 1964.A.J. 5. 989-1003. Svenska Lakartodningen. Incidence of Samonellae in clams. 27. U. 1984.. 1956. Applied and Environmental Microbiology. FDA Gulf Coast Technical Services Unit. seafood-handling procedures on Vibrio parahaemolyticus. Blake N. Sakazaki R. 1967. Effects of storage on microbial loads of two commercially important shellfish species.A. Wekell M.D. during depuration. Arcisz W.W. 32. P-1. 26. Jr. 22. Vaughn M. Food and Drug Administration. Food and Drug Administration. Food and Drug Administration. Journal of Food Protection. Fleet G..
97. 1989. Applied Environmental Microbiology. 1914. Journal of Bacteriology. West Coast States and British Columbia.. 38. 53. Urease-positive. Epidemiology and spectrum of Vibrio infections in a Chesapeake Bay community. Marine Fisheries Review. 45. 1937.. Kaysner C. 1147-9. Novak A. Obara Y. Kelly C.G. 27. Kelly M.T. Yokohama Medical Bulletin. Bacteriological control of oysters during processing and marketing.. 2820-2.. 107-8. Takikawa I. Jacobson M. 985-93. Bacteriological findings in Baltimore oysters. 39. Washington Report October 14. Loomis A. Vibrio parahaemolyticus illnesses. 273-80.W. Academic Press. Arcisz W. Ward W. 35. Akiyama S. 34. 1979. Joseph S. 1916. 46. Wekell M.G. Public Health Reports. Grodner R. 204-5. 1997. 1976. 133-47. 100. 1349-51.A. Crassostrea virginica..B.D. 41. Stroh E. Volume 1. Tanikawa E. Sample T. U. 36.... Serological and hemolytic characteristics of Vibrio parahaemolyticus from marine sources. Studies on pathogenic halophilic bacteria. kanagawa-negative Vibrio parahaemolyticus from patients and the environment in the Pacific Northwest.A.E. 2891-3. Virulent strains of Vibrio vulnificus isolated from estuaries of the United States west coast. Ed.M.. Kato T. Leitch J. 1958. Libonati J... Journal of Food Science.M.M.A. Journal of Clinical Microbiology. Zentralblatt für Bakteriologie. 1961. Janda J. 33.A. 60.. Journal of Infectious Disease. in Fish as Food.. 160. The bacterial flora of market oysters. Geiger J. Yamai S.. 42.S. 1954.. 1997. 1989.. (2).MOLLUSCAN SHELLFISH 32. Peeler R. Miyamoto Y. 13-7. Hoge C.. DePaola A.W.S. 353-7. Jr.. Journal of Bacteriology.. Joseph M. 37. Journal of Infectious Disease. 1989. 27.D.P.L. Morris J. The bacterial content of market oysters. 69. Geographic and monthly variation in composition of oysters. 38.C. Parasitenkunde. Abeyta C.. 128-31. 47.F.L. 25.. 1969. 2.. Fieger E. Journal of Clinical Microbiology. In vitro haemolytic characteristics of Vibrio parahaemolyticus: Its close correlation with human pathogenicity. Sidwell V. 1926.. Thompson C.. Stott R.N. Journal of Food Protection. Watsky D. 43. Shellfish Specialist. Vibrio infections associated with raw oyster consumption in Florida.. 41. 1. Bulletin of the Johns Hopkins Hospital. Kaysner C. Infectionskrankheiten und Hygien... Emergence of a restricted bioserovar of Vibrio parahaemolyticus as the predominant cause of vibrio-associated gastroenteritis on the West Coast of the United States and Mexico. 41. 40.. Ishibashi M. Berry F. Takizawa K. Jr. Bacteriological examination of oysters stored at low temperatures. 1987.M. Hlady W. Food and Drug Administration. Takeda Y. Microbiology of shellfish decomposition. Bothell.M. 73 .. 44. Vanderzant C. Abbott S.. Powers C. Israel E. Borgstrom G. 313-22. New York. U.
Hebert S.. Bradford H. Characterisation of Vibrios cholerae isolated from oysters.. 57. Jacobs R.E.. Applied and Environmental Microbiology. 45. 8.G. Barrett T. An epidemiological study of V.R. Jr.. Wekell M. Non-01 Vibrio cholerae bacteraemia: Case report and review.B. 343-50. Wazenski T.N.. Morris J.A.T..T...R. Hug M. Applied and Environmental Microbiology. 51.W. 1.. 275-83. McFarland L.G... Siebeling.A.. 59. Colwell R.W.G. Kaper J.. Caraway C..J.. Colwell R.M. Maden J.W.. Tacket C..... Safrin S. DePaola A. Non-O group 1 Vibrio cholerae gastroenteritis in the United States: Clinical. 49. DePaola A.A.. 37. Jr. Leitch J.T. Allegra D. Johnston J..A. 1983.. Roderick G.E. 53. Lockman H. Weaver R. 802-6. Plaard R.. Hug A.O. and enterotoxin production of Vibrio cholerae in Chesapeake Bay.A.J.B.. 1981. Wathen H.. Becker S. Roberts N. Ecology.. Presnell M.. Clinical features and an epidemiological study of Vibrio vulnificus infections.F.FISH AND SEAFOOD 48.A. 53. Craig J. 58.R. 1985.L.P.T. Microbial Ecology.J. Conte J. Review of Infectious Disease..G.O.. Adams M. Blake P. 61. Francis D.E..M. 558-61. Blake P. Duran W.. Tacket C. Davis B. 50. Blake P. Small P. Jr. Kaysner C. Wilson R.D. 46. 305-9. 312. Black R. Jr. Jr.G. 56. Non-01 Vibrio cholerae in shellfish. 299-312.. Applied and Environmental Microbiology. R. Puhr N.. 656-8. Kaper J.V.G. Bradford H. 52. Hunt J.M.. Journal of Infectious Disease.M. Journal of the American Medical Association. cholerae in Louisiana. Zywno S. sediment and waters of the U.C.. Wachsmuth I. 55. 1980.B. McFarland L.. 1475-8.. Blake P. Cholera and other vibrioses in the United States. McPhearson R.. Twedt R. Morris J. Peeler J. 1984. and laboratory characteristics of sporadic cases. 305.D. Stott R.. New York.A. Hollis D... 1982. Abeyta C.M. New England Journal of Medicine. Jr. Vibrios in the Louisiana Gulf Coast environment. 94.C. Jr. 1294-7. 1984. Cholera . Applied and Environmental Microbiology. McCay C. Morris J. Incidence of Vibrio cholerae from estuaries of the United States West coast. Ecological relationship between Vibrio cholerae and planktonic crustacean copepods..T. Wilson T. 1981.S.F. 54. in Vibrios in the Environment. 41. 91-103. 2050-3.G. Lee J.G.. Motes M...O. Ed.. Miller H..M. 60. Feldman R. Lancet.R. 1988. Brenner F.... West P. Vibrio vulnificus: Man and the sea...A. Spite G. 10. 149.M.. 5972.. 1344-8. 1987.A. Riddle C. Colwell R. 253. 1983.. Joseph S. New England Journal of Medicine.. epidemiologic. Snyder J. Journal of Food Protection. Twedt R.I. 1979. Morris J. 74 . 1982... Becker R.A possible endemic focus in the United States.M. Annals of Internal Medicine. Hickman F.F..E. Rahman R... serology. Pons V. 1985.W. Gulf Coast. John Wiley & Sons Inc. Illness caused by Vibrio damsela and Vibrio hollisae. 1012-7. Feeley J..M...K..
A.. 70.. Eds. Robinson D.E. 101-4. Levine M. 1980. Abeyta C. Phenotypic and genotypic characterisation of urease-positive thermophilic Campylobacter (uptc) isolated in N. Wareing D.. Wareing D. Journal of Infectious Disease. Dalley E. Wilson I.A. Murphy P. New York... 1993. Hunt J.MOLLUSCAN SHELLFISH 62. 1996.. Morris G.M..I. 1991.R.P. Austin S. 256. Jr.A. Blaser M. Huges T. Abeyta C. Part 1 Abstracts: Shellfish as a possible source of thermophilic Campylobacter spp. 1981. 621-4.... Alam A. Journal of Food Protection.. Jr. 4. Stott R. coli and lari) from shellfish and the marine environment. Disease from infection with Vibrio mimicus.A. Davis B. Black R.. A method for isolation of Campylobacter spp.G.. Helicobacters and Related Organisms. Clements M. Huq M.L. 51-64. The occurrence of Campylobacter jejuni and Campylobacter coli in Sidney rock oyster. Wilson T.R.A. Brussels. Moore J. Black R. 1584. 282. Journal of Rapid Methods and Automation in Microbiology.. Brenner D. C.E. Moore J. Ketley J. Campylobacter jejuni in a Washington State shellfish growing bed associated with illness. Eds. 67.J. Wekell M. Plenum Press. Incidence and survival of thermophilic campylobacters from shellfish growing water: media evaluation.K. Helicobacters and Related Organisms.A.M. 169-71. 64. S41.R.M.G. Kaysner C. Matsuda M.G. 472-9. infections in man..K. Ketley J.K..J.L...P..D. Public Health Laboratory Service.X... 1983.M. Wilson T.. Johnston J. 607-9... Kaysner.R.. London. Microbial Ecology in Health and Disease. Arumugaswamy R.W.S. 69. Experimental Campylobacter jejuni infection in humans. Jones D. in Campylobacters. Newell D. 65.G. Plenum Press. Deeter F.J. Clements M. Shandera W. Murphy P.S.J. in Campylobacter II: Proceedings of the Second International Workshop on Campylobacter Infections. Wilson I. 1988.A. 75 . 71. New York. from patients with diarrhoea..B. 6-9 September 1983. 157.H..A. Isolation of vibrio-like group. Feldman R. British Medical Journal.. 1987. Kaysner C.. (jejuni. Jr. Wekell M. 56. 68.G.M. 11. 63. in Campylobacters. Davis J.F.E. Studies of Campylobacter jejuni infection in volunteers.M.E. Pearson A. Newell D. 80.G. Eds. 1980. Griffin M. 1983.. 99.M... Fitzpatrick M.. Proudford R.. 13.. 73. Infective dose of Campylobacter jejuni in milk. 257..R. International Journal of Food Microbiology... 72. 66.. Rowe B. Abeyta C. Annals of Internal Medicine. Journal of the Medical Society of New Jersey. Blake P. Skirrows M..M.M. 323-5. Hughes T. 1996. 4. 4. Campylobacter gastroenteritis associated with raw clams. Journal of Clinical Microbiology.A. a newly recognised Vibrio species. Ireland. EF-6. Feldman R..G.D. Blaser M. 1995. Levine M..
Stott R. Farmer J. 76.N.. Burke V. Gracey M. Abeyta C. The use of bacterial membrane fractions for the detection of Campylobacter species in shellfish. Tenge B. Prevalence and distribution of Aeromonas hydrophila in the United States. 52.A. Aeromonas hydrophila: ecology and toxigenicity of isolates from an estuary.. vibrios and Aeromonas hydrophila. Annals of Internal Medicine. Kelly M. Robins-Browne R. Stelma G. Cooper M. 85. Recovery of Aeromonas hydrophila from oysters implicated in an outbreak of foodborne illness. 87.. Oyster borne disease outbreaks in St. 1997. Journal of Food Protection.. Report (1/13/84).A..M. 78. 1146-9. Tall B. Hood M. Koburger J. Partridge K.A.J..T. 998-1001. Kaper J. Wekell M. Kaysner C.. 52. 50. Hazan T. Incidence of motile aeromonads from United States west coast shellfish growing estuaries.. Beaman J. 27..R. Effects of processing and storing oyster meats on concentrations of indicator bacteria. epidemiology. 79.L..A. Holmerg S. Hickman-Brenner F.J. Atlanta.. Hirsch R. 1986. Colwell R.. Wekell M. and treatment of Plesiomonas shigelloides diarrhoea.. Lockman H.J. Department of Health and Human Services/Public Health Service/Food and Drug Administration. 1985.D. Miller M.F. 39. Indicator bacteria and Vibrionaceae multiplication in postharvest shellstock oysters. Journal of Clinical Microbiology. Infection and Immunity. 1989. 84. Robinson J. Baker R. 1988. Plesiomonas shigelloides: An opportunistic food and waterborne pathogen. 81. Petersburg. 55. Biotyping and virulence factors in clinical and environmental isolates of Aeromonas species. Blake P. 475-9. Applied and Environmental Microbiology. 47. 731-8. Journal of Applied Bacteriology. Bark D.. GA. Kaysner C. 449-57. Kain K.. 1987. Hunt J. 53. 598-601.L..C...A.D. 1990.A. 1980.M. Wachsmuth I... Applied and Environmental Microbiology. Journal of Food Protection. 76 . Herrington D. Journal of Food Protection. Seidler R. Kaysner C.. Singelton F. 77.M.M. Journal of Food Protection.. 5.P.J. Jr. 1989.. Journal of Food Protection.. Abeyta C.B. III. Journal of Rapid Methods and Automation in Microbiology.. Tzipori S. 1984.M. Levine M.. Frequent isolation of Edwardsiella tarda and Plesiomonas shigelloides from healthy Zairese freshwater fish: A possible source of sporadic diarrhoea in the tropics. Esch G. Van Damme L.K.. 1978..M... Cook D.. 343-9. 7-12. 849-55.C. Fliermans C. 80. Ruple A.. 86. Applied and Environmental Microbiology. Fla. 359-77. 1984..H. Trost P.A. 979-85. Abeyta C.R. 48. 82. 47.. 223-47.W. 83. 690-4. Jr. 105... Peterson D. Clinical features.W.. 36. Plesiomonas enteric infections in the United States. Jr. 1981.FISH AND SEAFOOD 74.A.L. Vandepitte J.. Wekell M. Sullivan J. In vitro and in vivo pathogenicity of Plesiomonas shigelloides.W.. Herrington T.D.. 75.M.
. fresh or frozenadulteration by bacteriological contamination Sec. in U. Gaithersburg.S. 2000. Washington D. Food and Drug Administration. Gould G. Burdge D. U. Baird-Parker T.. Martin R.600 (CPG 7119. Pittsurgh. 63-8.W.E..L. U. Collette R.H. U. 91.S. Ed. 560.W. Food and Drug Administration. Food and Drug Administration. Clams.8 Further Reading Ahmed F.S. Ed. 1987. Aspen Publishers Inc.S. 1995. Food and Drug Administration. U. 1997. 1991.C. Technomic Publishing Co.Pathogen monitoring of selected high risk foods (FY 88/89). in National Shellfish Sanitation Program Manual of Operations. 77 .R.A. U. Does Aeromonas hydrophila preferentially colonise the bowels of patients with hematologic malignancies? Diagnostic Microbiology and Infectious Disease.M.C. Food and Drug Administration. Food and Drug Administration.S. Seafood Safety. National Academy Press. Chapter 5: Foods.1987.. U.030. Fish Inspection. in U. oysters. 1995. Lund B. 90.S. Lancaster. Washington D.. Smith J. Government Printing Office . and HACCP: A Global Focus. Slavin J. Washington D.12). Ed. Sherlock C. Quality Control.C.E.MOLLUSCAN SHELLFISH 88. Food and Drug Administration Compliance Policy Guides. mussels.. The Microbiological Safety and Quality of Food. Inc.. Food and Drug Administration. 89. 7.S. Food and Drug Administration Compliance Program Guidance Manual... Part I Sanitation of Shellfish Growing Areas. 3.C.S. Food and Drug Administration. Program 7303.
4. and available in 0. lobsters. Shrimps. packed and frozen in 0. CRUSTACEAN SHELLFISH Linda Nicolaides.1 Definitions Crustaceans are invertebrate animals and represent a class of the phylum Arthropoda. jointed limbs and a chitinous exoskeleton. Species eaten from tropical waters include Penaeus spp. e. Such products are usually only partially cooked before freezing. and frozen in 1 and 2. Shrimp can be purchased in the raw or cooked state in a variety of forms: Entire shrimp. Crayfish.0 kg polythene bags. Pandalus borealis (deep-water shrimp). lobsters and crabs have ten legs. crayfish and shrimp (1). with additional water to form a glaze. peeled product..5 kilo pack (boxes or bags). and are usually aquatic. They possess a segmented body. Peeled ready-to-cook shrimp are raw. Parapeneopsis spp.g.5 and 1. and Trachypenaeus spp. crabs. FRSPH Ethical Trade and Food Management Group Natural Resources Institute University of Greenwich Central Avenue Chatham Maritime Kent ME4 4TB United Kingdom 4.0 kg polythene bags. 79 . with shells and heads on or off are usually packed.Ph. (deep-water shrimp). Peeled and cooked shrimp are ready-to-eat after controlled thawing. prawns and scampi are temperate species commonly eaten by man and include members of the families Nephrops norvegicus (scampi).. Breaded shrimp and scampi are battered and coated in breadcrumbs and sold frozen to the consumer for cooking prior to consumption. M. Pandalus montagui and Crangon crangon (inshore shrimp)..5 and 1. the front pair ends in claws.
packed into polythene bags. Following capture or harvest. spoilage bacteria are of little importance as. However. e. Microbiological risks associated with crustacean shellfish increase in proportion to the degree of handling that the product undergoes. lobsters and crayfish are traditionally kept alive after capture and then cooked to kill immediately before consumption. Poor hygienic procedures will increase the probability of contamination with bacteria of public health significance. e. hence. their feeding and living habits. only live carcasses are processed. so that spoilage may occur during landing and transportation to the processing plant. 4. for crabs and lobsters. which means that the haemolymph can be a reservoir of bacteria. The structure of the circulatory system of crabs is not closed. tropical waters. The microbial flora of crustaceans from temperate waters differs from that of crustaceans from warmer. Cooked and chilled. Spoilage begins following death once the immuno-response system of the crustacean shellfish fails to function. particularly after it has been cooked. traditionally..FISH AND SEAFOOD Crabs.2 Initial Microflora As with finfish. If these products are subjected to temperature abuse. boiled to kill the crustaceans and cook their meat prior to consumption. there is a growing market for fresh lobster tails or cooked crabmeat in crab shells. lobster and crayfish are available for the consumer owing to the growing demand for ready-to-eat products. Shewanella putrefaciens and 80 . Control measures to prevent these potential hazards occurring include training and supervision of operatives as well as Good Hygienic Practices (GHP). Pasteurised crabmeat is edible meat removed from the exoskeleton by hand. In comparison. the level and type of the initial microflora of crustacean shellfish will reflect a combination of factors. the geography of the area they are captured from or are grown in. lobster tails and cooked crabmeat presented decoratively in their shells. Spoilage bacteria associated with temperate crustaceans include Pseudomonas spp. stored and transported as frozen product. it is important that shrimp are handled and iced correctly in order to keep them in as fresh and uncontaminated a state as possible. the season. which include the environment from which they have been captured or harvested. shrimp die immediately after capture. during the peeling of cooked shrimp. or frozen crab. particularly for members of the genus Vibrio (2). storage above the recommended storage temperature. the flora will change depending upon the methods of handling and/or the environmental conditions to which they are exposed. As with finfish. For example.g. and the temperature and quality of the waters in which they exist. with the increasing demand for ready-to-eat/cook crustacean products.g. this presents more favourable conditions for growth by both spoilage and pathogenic bacteria. the potential exists for crosscontamination from both the processor and from the environment.
by removing the head and thorax a source of enzymes is eliminated that will prevent the occurrence of black spots.10 days for temperate water species (3).. 4. or cooked. from the time of capture and processing. Scrupulous hygiene is important at this stage to prevent the cross-contamination of the shrimp tails. however. In comparison shrimps die quickly after capture. Shrimp might be further processed with the shell on. As with finfish. Hence. peeled and cooked. there is evidence that the shelf life of tropical crustaceans is longer than that of temperate-water species stored correctly in ice .3. glazed and frozen (see Figure 4.. 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. up to 16 days for tropical species compared with 8 . Shrimp will then be glazed and frozen in a raw state. In contrast.1. chilled sea water. the genera associated with tropical species include Enterobacteriaceae. graded according to size. or with the shell and central vein removed. A large proportion of shrimps are produced by aquaculture. uncooked prawns and shrimp is presented in Figure 4. After capture and transport to the processing plant. Micrococcus spp. whether GHP are in place. the rate of deterioration of the crustacean shellfish by these organisms is reduced. 4. Alcaligenes.. They are then taken to market alive in either holding tanks. bacterial contamination and spoilage are inevitable. When the shrimp arrive at the processing plant they are washed and then sorted into size. The claws are usually bound together to prevent the animals fighting and causing damage to each other. The rate of spoilage will be a function of time and temperature. or transported directly to processing plants. The latter process can expose the shrimp muscle to incidents of cross-contamination if GHP are not followed. especially if they are stored in ice or iced water. with “head on” or “head off” whereby the edible tail is removed from the head. As the muscles of crustacean shellfish decompose rapidly after death dead animals are rejected.1). These animals can survive out of water if the surrounding environment is kept cool and damp.CRUSTACEAN SHELLFISH members of the Acinetobacter-Moraxella group. or are found in brackish water around the shores. so they are either sold on the local market or are processed in local processing plants. and will be dependent upon whether the crustaceans were harvested from cool or warmer waters. Pseudomonas spp.3 Processing and its Effects on the Microflora Crabs. corynebacteria.for example. thus. and Achromobacter spp. gills and thorax. Shrimps can be processed whole. they are washed.1 Prawns and shrimps An example of the process used for preparing frozen. chilled 81 . Vibrio spp. lobsters and crayfish are harvested from cages or pots containing bait.
4. particularly the pathogens Bacillus cereus and Clostridium perfringens originating in the flour. Good Manufacturing Practice (GMP).1. including GHP.frozen uncooked prawns 82 . should be followed to ensure that the product is not re-contaminated with microorganisms present in the processing environment or from the workers. After cooking. spices and cooking batter. Flow diagram .FISH AND SEAFOOD frozen. and packed to meet the customer’s specification. Fig. The cooking stage will eliminate the intrinsic microbial flora of the shrimp as well as any contaminants that have been added during handling and transportation to the processing plant. Breaded products introduce an added risk of spore forming bacteria.
The shell will be soft. for crabs. An offensive odour is also produced. If the tail drops. the meat is either repacked and sold in the shells. However. or in hermatically sealed cans that have undergone an appropriate heat treatment.3. Once removed and cleaned. there is a new hazard of contamination of the cooked meat by workers during its physical removal from the exoskeleton. If boiled. With spoiled crabs and lobsters. 83 . the tightness of limbs will indicate whether they were killed by immersion in boiling water or. Heat is produced if the shrimps decompose in a confined space. In comparison. since crabs and lobsters are traditionally consumed immediately after cooking.CRUSTACEAN SHELLFISH 4. slightly iodine smell. wet and ‘soapy’ or ‘jammy’ to the touch.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.2 Cooked crabs and lobsters As indicated earlier. The shells have a crisp. bright and dry appearance. the joint between the tail and carapace is open and the limbs hang loosely. 4. or packaged and processed. putrefaciens. indicative of protein degradation and the growth of the principal spoilage bacterium Sh. with the growing demand for ready-to-eat products from crab and lobster meat. contaminating microflora may include enteric bacteria. they include Sh. 4. and a strong ammoniacal odour is produced. there is no indication of life. Processed crab and lobster meat can also be bought as packages of meat that have been subjected to a pasteurisation step. with visible decomposition or green discoloration seen through the membrane exposed by the dropping tail. putrefaciens and Pseudomonas spp. viruses or parasites. immediately prior to boiling. whilst boiled crustaceans have a pleasant slightly sweet smell. 4. If hygienic procedures are not followed. there is no associated spoilage problem.4. and have a sweetish.4. spoiling prawns and shrimps have shells that are soft.2 Crabs and lobsters Crabs and lobsters should be alive when boiled or sold. bacteria of public health significance. dull and sticky.1 Prawns and shrimps Freshness indicators for prawns and shrimps are for the shells to be crisp and dry and feel sharp and cool to the touch.
monocytogenes was absent from 40 samples of cooked prawns. in domestic/imported shrimps (15). nearly 30 years elapsed before the potential hazard of L. shrimp and shellfish (11). often contains Salmonella and other enteric pathogens (8) and has been demonstrated to contaminate crustaceans growing in these environments. the first in a series of Class I recalls was issued in May 1987 to retrieve nearly 4 tonnes of tainted crabmeat (11). which required automatic detention of all Mexican frozen crabmeat that tested positive for Listeria and Escherichia coli (12).5 4. Hence. 7). Moreover. Salmonella cannot be completely removed from raw shrimp during processing. Although foodborne illness had not been associated with the product. in compliance with the Food and Drug Admistration’s (FDA) ‘zero tolerance policy’ for L. fish fingers. Further surveys identified Listeria spp.5. shrimps and cockles sold (17). 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. animal manure. 16).5% of frozen raw shrimps from different countries (13). monocytogenes in contaminated crustacea was recognised in the United States of America (USA) when the pathogen was isolated from Mexican frozen cooked crabmeat (10).1 Pathogens: Growth and Survival Salmonella spp. a common pond fertiliser used in aquaculture. Following this. 4.FISH AND SEAFOOD 4. although efficient cooking will quickly eliminate it (6). 6.cooked prior to consumption. One of the earliest accounts of the presence of Listeria monocytogenes in crustaceans was by Russian workers in 1959 (9). However. 5. cooked and smoked fish. Results of a survey carried out in England and Wales between 1987 and 1989 demonstrated that L.2 Listeria spp. The above results prompted the FDA to develop a compliance programme for the pathogen. The presence of L. even if GMP is followed. as well as for Salmonella. They are mesophilic microorganisms usually residing in the intestinal tracts of human beings and warm-blooded animals. The pathogen was later identified in frozen raw shrimp (13) and lobster tails (14) . in 24. and to increase testing of many other domestically produced seafoods under the General Pathogen Surveillance Programme (14. monocytogenes in cooked ready-to-eat foods. The genus Salmonella contains a wide variety of ‘species’ pathogenic for man or animals. later surveys in 1990 recovered the pathogen from retail raw. An import alert was put in place in June 1987.5. and usually for both. Aquatic birds and the hands of workers have also been identified as sources of the pathogen. monocytogenes in cooked ready-to-eat crustaceans is recognised as a potential risk to selected groups of individuals: the very young and 84 .
is widely distributed in aquatic environments. pasteurisation and cooking. was associated with seafood consumption. cholerae is divided into two groups. the pathogen can grow at temperatures as low as 3 °C. aureus on their hands. so is able to survive heat treatments that eliminate vegetative cells. whilst boils and cuts are also common sources of the pathogen.g. V. botulinum type E at 10 °C but not at 4 °C (20). especially in products that have been vacuum packaged. combined with the degree of handling of these types of product. e. V. Vibrio parahaemolyticus and Vibrio vulnificus are species that have been reported to be associated with raw crustaceans harvested from estuarine waters (21.3 Staphylococcus aureus Staph. cholerae O1 and V. cholerae. 19). 24. no incidence of foodborne illness caused by L. aureus is not a member of the intrinsic flora associated with crustacean shellfish. although more recently the non-O1 type has also been demonstrated to cause foodborne illness in crustaceans. The former. The pathogen has resistant spores. To date.CRUSTACEAN SHELLFISH old.5. Crustaceans have been linked with food intoxications attributed to Staph. monocytogenes has been attributed to crustaceans. An epidemic that began in South America in 1991. 4. Once the competing microflora has been removed by a heat treatment. The main source of contamination is the hands of processing personnel. 22) as well as from undercooked crustacea when harvested from contaminated waters (23. 85 . particularly crabmeat (18. Control should be achieved by promoting GHP and staff training programmes. although contamination by types A and B may occur from the environment. the causative agent of cholera. pregnant women and the immuno-compromised. therefore. Crabs are covered in a chitin exoskeleton.4 Clostridium botulinum C. cholerae non-O1.5. 25). GMP should be followed in the processing plant to ensure that the final product is held at 3 °C. 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. However. 4.5 Vibrio spp. Vibrio cholerae. Researchers have demonstrated that inoculated shrimp supported toxin production by C. which was eventually attributed to a ship discharging its bilge into the coastal waters of Peru. 4. botulinum type E is commonly associated with fish and fishery products. about 50 .5. as part of the natural bacterial flora. which can become colonised by V. aureus because of the ubiquity of this organism in the processing environment.70% of healthy individuals carry toxin-producing strains of Staph.
non-B Hepatitis viruses (31). been implicated in a case of foodborne infection associated with crustaceans.9 Viruses Crustaceans grown in waters contaminated with sewage may become carriers of viruses such as Hepatitis A. Norovirus. however. particularly trematodes (6). South America. e. parahaemolyticus. 4. whilst 5. or freezing at a temperature of -23 °C for 7 days (30. 32). sobria are known to be fish pathogens and are commonly isolated from ponds used for aquaculture. 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.30 MPN/g) (27). but has the potential to cause fatal foodborne illness (26).8% of samples contained C.2 °C) so that GHP coupled with controlled low temperature storage is essential to prevent the pathogen from reaching levels that may present a hazard. V.6 Campylobacter jejuni and Campylobacter coli C. However. although A. in a survey carried out in the United States between 1977 and 86 . Japan. a marine halophile. However. A. Of these. V. the potential hazard exists as C. jejuni has not.8 Parasites The use of animal excreta in aquaculture has also been associated with foodborne parasitic infections of crustaceans. 4. The organism is widespread in estuarine waters and is a common isolate from harbour water in warm climates.7 Aeromonas hydrophila and Aeromonas sobria A. Quantitative levels were below limits of detection in all cases (<0.g.5.5. 4. Hepatitis A virus presents the most serious hazard. hydrophila has been isolated from prawns (29).FISH AND SEAFOOD and caused major economic losses to South American countries that export shrimp to the United States.5. jejuni was isolated from 36 (15%) of 240 samples of freshly hand-picked blue crab (Callinectes sapidus) meat. they have not been confirmed to be causal agents of foodborne illness associated with crustaceans (28). to date. Astroviruses and non-A. 4. coli. hydrophila is able to grow well at refrigeration temperatures (0 . It is more commonly implicated in countries where crustacean shellfish are eaten raw.5. Parasitic infections are controlled by either cooking infected material prior to consumption. vulnificus is an invasive and lethal pathogen associated with wound infections. occurs in crustaceans from warm environments. hydrophila and A. Caliciviruses. 31.
Norwalk viruses can also withstand heating to 60 °C. 4. which results in a persistent and apparently permanent loss of short-term memory (36). Therefore any microbiological criteria need to be part of this HACCP-based system to verify that the system is under control. or chlorine concentrations similar to those found in drinking water. although it is one of the European Union (EU) legislated allergens. Selected parts from this regulation is presented below.6.47% of outbreaks attributed to Hepatitis A virus were attributed to a foodborne origin (33). A summary of microbiological criteria for crustacean shellfish is listed below.10 Allergens Allergens have not been reported to be a significant hazard with crustacean shellfish. produced or imported into the European Union (EU).1 EU Legislation Microbiological standards for cooked crustaceans and shellfish. Provisions laid down concerning the use of additives would also apply to these types of products.CRUSTACEAN SHELLFISH 1981. 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. The risk of such hazards is far higher with fin fish and molluscan shellfish.6 Published Microbiological Criteria The global legislative requirement for controlling the safety (and quality) of crustacean shellfish is to apply a preventive. 4. 4. Both Hepatitis A virus and Norovirus are stable pathogens and are relatively resistant to inactivation (34). 38. only 0. 4. are included in the Commission Regulation (EC) No.5. from farm to fork (37. consistent with that in the stomach. 87 . No foodborne incidents have been attributed to this source to date. The toxin causes amnesic shellfish poisoning (ASP) when consumed. 39).5. which could be transferred to the meat during unskilled removal from the shell (35). They can survive low pH levels.11 Chemical pollutants and biotoxins Researchers have demonstrated that crabs feeding in waters contaminated with algae do have potentially high levels of algal toxin domoic acid (DA) in their viscera. 2073/2005.
FISH AND SEAFOOD
220.127.116.11 Microbiological criteria applicable to the production of cooked crustaceans (Commission Regulation (EC) No 2073/2005 on microbiological criteria for foodstuffs) 18.104.22.168.1 Chapter 1 - Food safety criteria
L. monocytogenes Prepared crustacean products fall into the category described in the regulation as “Ready-to-eat foods available to support the growth of L. monocytogenes other than those intended for infants and for special medical purposes”. During capture and preparation there is a zero tolerance for the presence of L. monocytogenes on the product. However, to take into account the issues of natural habitats being contaminated with low levels of this pathogen there is a slight loosening of the regulation at the point of sale and for the duration of the products shelf life (refer to Table 9.I, Food Category 1.2). If the level of L. monocytogenes should be above this level then the consignment is considered to be unsatisfactory. Salmonella All cooked crustaceans and molluscan shellfish should be free from Salmonella at the point of sale and for the duration of their shelf life (refer to Table 9.I, Food Category 1.16). Should any sample test positive for Salmonella then the product is declared to be unsatisfactory. 22.214.171.124.2 Chapter 2 - Process hygiene criteria
Process hygiene criteria for E. coli and coagulase-positive staphylococci in shellfish and shucked products of cooked crustaceans and molluscan shellfish are laid down (refer to Table 9.II, Food Category 2.4.1). 4.6.2 International requirements
International requirements are based upon having a risk-based, preventive management system in place at all stages of the supply chain. The requirements for this preventive approach is described as follows (37): • Fish and fishery products should be prepared in plants certified by the local competent authority. All certified plants should comply with the GHP (39); • The fisheries industry should take responsibility, implement and maintain safety management systems based upon HACCP; • The national competent authority is responsible for the certification of fish processing and manufacturing plants, verification of effective systems, and
issuing of certificates of compliance for export products. This includes auditing and inspection programmes; • National surveillance and monitoring programmes should be in place to demonstrate that all identified hazards are under control, e.g. biotoxins, and to identify potentially emerging hazards; Within this requirement is the flexibility for countries and trading blocks to set specific standards required to deliver an equivalent requirement with those at national level. Such standards need to be science-based, especially if they are more stringent that those recommended by the Codex Alimentarius Commission. Verification that GHP have been followed during the handling and preparation of crustacean shellfish for consumption is usually done by assessing the level of E. coli and/or coagulase-positive staphylococci in the products. Legal requirements generally require that these groups of indicator organisms are present in levels <100 cells/gram of product. Pathogenic bacteria such as Salmonella spp. should be absence is 25 grams multiplied by the number of subsamples tested. 126.96.36.199 United States The FDA brought into force a Federal Mandate Seafood Rule in 1995 which forms the foundation for good practices required for the handling and processing of imported fish and fishery products into the U.S. The best practices include GHP and HACCP (37). All fish and fishery products produced in and imported into the U.S. after 17 December 1997 are required to demonstrate that HACCP principles were applied to their handling and processing. The USFDA has a zero tolerance for Salmonella in fish and fishery products; this includes farmed shrimp. The US FDA considers that the presence of pathogenic microorganisms in seafood is an indicator of adulteration (32). 4.7
Encarta 1997. Encarta 97 Encyclopedia World English Edition. Microsoft. The International Commission for Microbiological Specifications for Food. Fish and Fish Products, in Microorganisms in Foods 6. Microbial Ecology of Food Commodities. Ed. The International Commission for Microbiological Specifications for Food. New York, Kluwer Academic/Plenum Publishers. 2005, 174-249. Cann D.C. Bacteriology of shellfish with reference to international trade, in Handling, Processing and Marketing of Tropical Fish. Ed. Tropical Products Institute. London, Ministry of Overseas Development. 1976, 377-94. D’Aoust J.Y., Gelinas R., Maishment C. Presence of indicator organisms and the recovery of Salmonella in fish and shellfish. Journal of Food Protection, 1980, 43, 679-82.
FISH AND SEAFOOD 5. 6. 7. 8. 9. 10. 11. Fraiser M.B., Koburger J.A. Incidence of salmonellae in clams, oysters, crabs and mullet. Journal of Food Protection, 47, 343-5. Reilly P.J.A., Twiddy D.R., Fuchs R.S. Review of the occurrence of Salmonella in tropical shrimp. FAO Fisheries Circular No. 851, 1992, (FIIU/C851). Reilly P.J.A. Salmonella and Vibrio cholerae in brackishwater cultured tropical prawns. International Journal of Food Microbiology, 1992, 16, 293-301. Twiddy D.R. Antibiotic-resistant human pathogens in integrated fish farms. ASEAN Food Journal, 1995, 10, 22-9. Olsufjew N.G., Petrow V.G. Detection of Erysipelothrix and Listeria in stream water. Zhurnal Mikrobiologii Epidemiologii i Immunobiologii, 1959, 30, 89-94. Anon. First Listeria finding in crabmeat confirmed by FDA. Food Chemistry News, 1987, 29 (14), 38. Ryser E.T., Marth E.H. Incidence and behaviour of Listeria monocytogenes in fish and seafood, in Listeria, Listeriosis and Food Safety. Eds. Ryser E.T., Marth E.H. New York, CRC Press. 2007, 617-54. Anon. Mexican crabmeat, Greek pasta automatically detained. Food Chemistry News, 29 (19), 43. Anon. FDA checking imported domestic shrimp, crabmeat for Listeria. Food Chemistry News, 1987, 29 (24) 15-7. Archer D.L. Review of the latest FDA information on the presence of Listeria in foods. WHO Working Group on Foodborne Listeriosis, Geneva, Switzerland. 1988, 15-9. Anon. FDA to sample shrimp for Salmonella, Listeria. Food Chemistry News, 1988, 30 (36), 9-10. Anon. Monitoring of changes to listeriosis problem urged. Food Chemistry News, 1989, 31 (20), 13-7. Richmond M. Report of the Committee on the Microbiological Safety of Food. London, HMSO. 1990, 133-7. Swartzentruber A., Schwab A.H., Duran A.P., Wentz B.A., Read R.B. Jr. Microbiological quality of frozen shrimp and lobster tail in the retail market. Journal of Food Protection, 1980, 40, 765-9. Wentz B.A., Duran A.P., Swaartzentruber A., Schwab A.H., McClure F.D., Archer D., Read R.B. Jr. Microbiological quality of crabmeat during processing. Journal of Food Protection, 1985, 48, 44-9. Garren D.M., Harrison M.A., Huang Y-W. Clostridium botulinum type E outgrowth and toxin production in vacuum-skin packaged shrimp. Food Microbiology, 1994, 11, 467-72. Molitoris E., Joseph S.E., Krichevsky M.I., Sindhuhardja W., Colwell R.R. Characterisation and distribution of Vibrio alginolyticus and Vibrio parahaemolyticus isolated from Indonesia. Applied and Environmental Microbiology, 1985, 50, 1388-94.
12. 13. 14.
15. 16. 17. 18.
CRUSTACEAN SHELLFISH 22. Varma P.R.G., Iver T.S.G., Joseph M.A., Zacharia S. Studies on the incidence of Vibrio cholera in fishery products. Journal of Food Science and Technology, 1989, 26, 341-2. Madden J.M., McCardell B.A., Morris J.G. Jr. Vibrio cholerae, in Foodborne Bacterial Pathogens. Ed. Doyle M.P. New York, Marcel Dekker. 1989, 525-42. Twedt R.M. Vibrio parahaemolyticus, in Foodborne Bacterial Pathogens. Ed. Doyle M.P. New York, Marcel Dekker. 1989, 543-68. Oliver J.D. Vibrio vulnificus, in Foodborne Bacterial Pathogens. Ed. Doyle M.P. New York, Marcel Dekker. 1989, 569-600. Pearson A.M., Dutson T.R. Quality Attributes and Their Measurement in Meat, Poultry and Fish Products. Advances in Meat Research Series. Volume 9. London, Chapman & Hall. 1994. Reinhard R.G., McAdam T.J., Flick G.J., Croonenberghs R.E., Wittman R.F., Diallo A.A., Fernandes C. Analysis of Campylobacter jejuni, Campylobacter coli, Salmonella, Klebsiella pneumoniae and Escherichia coli 0157:H7 in fresh handpicked blue crab (Callinectes sapidus) meat. Journal of Food Protection, 1996, 59 (8), 803-7. Buchanan R.L., Palumbo S.A. Aeromonas hydrophila and Aeromonas sobria as potential food poisoning species: a review. Journal of Food Safety, 1985, 7, 15-29. Palumbo S.A., Maxino F. Williams A.C., Buchanan R.L., Thayer D.W. Starch ampicillin agar for the quantitive detection of Aeromonas hydrophila in foods. Applied and Environmental Microbiology, 1985, 50, 1027-30. Wekell M.M., Manger R., Colburn K., Adams A., Hill W. Microbiological quality of seafoods: viruses, bacteria and parasites, in Seafoods: Chemistry, Processing Technology and Quality. Eds. Shahidi F., Botta J.R. Glasgow, Blackie Academic and Professional. 1994, 196-219. Schantz J., McKerrow J.H. The dangers of eating raw fish. New England Journal of Medicine, 1989, 320, 1143-5. Ahmed F.E. Seafood Safety. Seafood Safety. Committee on Evaluation of the Safety of Fisheries Products, Food and Nutrition Board, Institute of Medicine. Washington D.C., National Academy Press. 1991. Blackwell J.H., Cliver D.O., Callis J.J., Heidelbaugh N.D., Larkin E.P., McKercher P.D., Tayer D.W. Foodborne viruses: their importance and need for research. Journal of Food Protection, 1985, 48, 717-23. Kapikian A.Z., Chanock R.M. Norwalk group of viruses, in Virology. Eds. Fields D.M., Knipe D.M. New York, Raven Press. 1990, 359-77. Horner R.A., Postel J.R. Toxic diatoms in western Eashington waters (US west coast). Hydrobiologia, 1993, 269/270, 197-205. Perl T.M., Bedard M.S.M., Kosatsky T., Hockin J.C., Todd E.C.D., Remis R.S. An outbreak of toxic encephalopathy caused by eating mussels contaminated with domoic acid. The New England Medical Journal, 1990, 322, 1775-80.
23. 24. 25. 26.
34. 35. 36.
FISH AND SEAFOOD 37. Ababouch L., Gandini G., Ryder J. Causes in Detentions and Rejections in International Fish Trade. FAO Fisheries Technical Paper 473. Rome, Food and Agriculture Organisation of the United Nations. 2005. Australian Government. Australian Quarantine and Inspection Service Hazard Analysis and Critical Control Point (HACCP). A Guide to Compliance with Export Control (Fish and Fish Products) Orders 2005. FAO/WHO. General Hygiene Requirements. Rome, Food and Agriculture Organisation of the United Nations. 2003.
Further Reading Spoilage and pathogens
Gram L., Huss H.H. Fresh and Processed Fish and Shellfish, in Microbiological Safety and Quality of Food Volume 1. Eds. Lund B.M., Baird-Parker A.C., Gould G.W. Maryland, Aspen Publishers Inc. 2000, 472-506. Ababouch L., Gandini G., Ryder J. Causes of Detentions and Rejections in International Fish Trade. FAO Fisheries Technical Paper 473. Rome, Food and Agriculture Organisation of the United Nations. 2005. International Commission on Microbiological Specifications for Foods. Fish and fish products, in Microorganisms in Foods 6. Microbial Ecology of Food Commodities. Ed. International Commission on Microbiological Specifications for Foods. London, Blackie Academic and Professional Press. 1998, 130-89. Sutcliffe P., Disney J. Handling, Processing and Marketing of Tropical Fish. London, Tropical Products Institute. 1977. Connell J.J. Control of Fish Quality. Oxford, Fishing News Books. 1995 Ward D.R., Hackney C. Microbiology of Marine Food Products. New York, Van Nostrand Reinhold AVI. 1991 Pearson A.M., Dutson T.R. Quality Attributes and their Measurement in Meat, Poultry and Fish products. Advances in Meat Research Series. Volume 9. London, Chapman & Hall. 1994.
4.8.2 Pathogens - general
International Commission on Microbiological Specifications for Foods. Microorganisms in Foods 5. Characteristics of Microbial Pathogens. Eds. International Commission on Microbiological Specifications for Foods. London, Blackie Academic and Professional Press. 1996. Varnam A.H, Evans M.G. Foodborne Pathogens - An Illustrated Text. London, Wolfe Publishing Ltd. 1991. Doyle M.P. Foodborne Bacterial Pathogens. New York, Marcel Dekker. 1989.
rather than for preservation itself. and a preliminary operation to some smoking.5. and before significant spoilage takes place. Fish can be salted in three different ways or their combination: Dry salting. The immersion in concentrated brines for long periods is usually used for fatty species and for longer-term preservation. dependent on the country or the type of product. This can be called ‘kench’ curing and is mainly used for non-fatty white fish.1 Definitions Curing is the preservation of fish by removing or displacing available water in the fish flesh to a level that discourages microbial growth. their combinations and/or variations. The fish is immersed in a brine solution in which different salt concentrations can be used according to the process. split and opened out flat. The processes take anywhere from a few hours to a few days or even months. Simon Derrick The Grimsby Institute Humber Seafood Institute Europarc Grimsby DN37 9TZ United Kingdom 5. both to preserve and to create a variety of new products. and salting is done by stacking the fish in layers. The fish is headed. where the salt replaces moisture in the fish flesh. Salting is both a method of preserving fish. and is sometimes known as ‘pickling’. It includes the salting. to assure a salt concentration in the fish flesh that is appropriate for flavouring purposes prior to the use of other preservation techniques. Fish curing is practised worldwide. fish is immersed for a few hours in a relatively low concentration brine. Wet salting or brining. SMOKED AND DRIED FISH Dr. and marinating processes. 93 . Usually. alternating fish layers with a layer of salt. and the extracted moisture is allowed to drain away. CURED. drying. drying and smoking processes.
this ‘brine’ can be collected and placed again over the fish several times. Usually. Salt. thus making the product unsuitable for microbial growth. Freeze drying. Thus. and the air movement removes the vapour. usually vinegar. depending on the type of product. Heat is transferred to fish from heated air or a heated surface. Marinating or pickling. is used as a preservative instead of salt. 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 . Food products can be dried in three different ways: Air or contact drying. it can also be named ‘pickling’ or ‘blood pickling’. the preservative effect is due to the low pH used in the process. There are some marinating processes where salt is used for the first cure and a firmer product is obtained. When the process is running in a closed system. The principle of preservation of the first method is the inactivation of enzymes and killing the non-spore-forming organisms. without other preservation methods applied after boiling. Drying is the preservation of fish by removing the moisture content of the fish flesh by evaporation. Included in this group are some lightly preserved products where no acid is added to the marinade.FISH AND SEAFOOD Dry / wet salting. evaporated water is removed by vacuum pump. Marinades are acidified brines. The product is dried by contact with heated surfaces or by radiation. This is a mixture of a dry and a wet salting process. are added to the marinade. thus delaying spoilage for a few days. It is not commonly used for fish products. prior to or during the boiling process. The fish is mixed with dry salt but in a watertight container. Salted-boiled fish are the products obtained on either boiling fish in a brine. as concentrated brines are formed. and the fish can be immersed in such brines for long periods. Smoking is an ancient process for preserving and flavouring food. Salt-boiling. or boiling it with dry salt and water. such as ‘gravad’ fish. Longer shelf life is achieved in the products when salt is added. After the freezedrying process. under normal pressure. the ice sublimes and the vapour is removed from the fish by vacuum pumps. thus reducing the water activity of the product. The products are usually consumed raw. As the water is drawn out of the flesh by the salt a brine is formed. the product rehydrates quickly but does not retain reabsorbed moisture on cooking. Vacuum drying. as well as other flavourings (sugar. It has found limited application with fish. where an organic acid. this was done naturally by exposure of the fish to the air. Fish in contact with refrigerated plates freeze. spices). This is the common method of drying fish. Traditionally.
The secondary objective is to impart sensory characteristics such as smoke colour and smoke flavour to the product. or by means of smoke flavouring processes. In this process. including pathogens. A dry or wet salting step is always used in the process before exposure to the wood or sawdust smoke. smoking is now used mainly to give the fish a characteristic flavour. With the exception of some Asian countries. or liquid smoke. 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. The different spices used in the immersion brines are also a potential source of microorganisms. The smoke itself may contain spores of moulds. In this process. Cold-smoking.2 Initial Microflora The initial microflora are those of the raw fish from which the products are made. Also. by exposure to the wood or sawdust smoke. Hot-smoking. which irreversibly combine with the haemoglobin in the tissue to maintain the red colour. such as injection or dipping in a solution of liquid smoke. 95 . It is commonly used in place of traditional smoke. the fish is subjected to heat for a sufficient period of time to coagulate protein throughout the fish. process temperatures are over 60 °C. or by means of smoke-flavouring processes with liquid smoke. and the preservative effect is only slight. leaving only elevated levels of carbon monoxide. Liquid smoke is a natural aqueous condensate of wood smoke that has been aged. Smoked-flavoured fish. The process was designed so that the smoke is filtered to remove all smoke particles. Clear smoke. the salt used in the processes may contain halophilic bacteria. SMOKED AND DRIED FISH of smoke. or added to the brine with other flavourings. by exposure to the wood or sawdust smoke.CURED. However. sporeforming bacteria or osmophilic moulds. fish can be smoked by means other than the direct action of smoke. Smoked fish is prepared by subjecting the fish to the direct action of smoke from wood or wood sawdust burning. Smoking can be either traditional. the sugar used in some products may contain yeasts and moulds as well as spore-forming bacteria. 5. It can be smoked either traditionally. which impart the flavour of smoke to fish. After the salting step. the fish is subjected to heat for a period that does not coagulate protein. Usually. It can be injected into the product. Usually. temperatures below 30 °C are used.
When a milder brine.8 211. a 0º brine saturation means pure water. A 100º brine solution means that the solution is saturated and no more salt can be dissolved. all using salting steps. TABLE 5. Salt concentration means the percentage of salt in the water contained in the fish. processes and producers’ private recipes.4 52.0 g of salt per litre of brine). it causes swelling because the fish absorbs water from the salt solution (1).6 g of salt or more per litre of brine) may cause salt burn in this way. smoking or marinating processes. % 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. which is the percentage of salt in the whole weight of fish that is water plus protein plus fat.8 211.6 132. it is impossible to establish standardised processing stages in the curing of fish. or salt in the water phase. it is important to state the temperature. The dissolution of salt in water is dependent on the temperature.3 Processing and its Effects on the Microflora Owing to the wide variety of products.8 79. When a 100º brine is used. This should be distinguished from the salt content. it is important to understand the concept of salt concentration. and salt content. When working with salting.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. about 50º (132.4 184.FISH AND SEAFOOD 5. is employed.0 .6 264. Different brine strengths are required according to the curing processes and/or the type of product.0 158.I). the fish is usually salted with a 70 .2 g of salt per litre of brine).4% salt (Table 5. a 90º or above brine (237.2 105. etc. The strength of the solution is described as degrees of saturation (brineometer degrees). For smoking salmon. Salt in the water phase is the amount of salt compared with the total amount of water and salt in the fish.2 237. Thus.80º brine (184. sometimes the salt crystallises onto the surface of the fish. Therefore. Salt in the water phase links the two conditions of moisture and saltiness. only the most common processes can be described here. At 18 °C a 100º brine is produced when the solution contains 26.
1 Salting Salting of fish for preservation purposes has been used since pre-history. The most common fish used in the northern hemisphere is cod (Gadus morhua). outgrowth of halophilic bacteria may occur. are recommended to help make the product whiter. and a ‘putty’ fish product is also obtained (1). however.CURED. lean white fish is used for this purpose.3. For a light salting.35% and 0. caught in North Atlantic waters.1 Dry salting / Kench curing Usually.0. which include calcium and magnesium. and such delays. can lead to the decomposition of the centre of thick muscles before it reaches the level of salt concentration sufficient to prevent spoilage. A ‘green cured’ product is obtained when the water content has been reduced to about two-thirds of the initial amount.1 ppm and 10 ppm. for which there is a high demand in Mediterranean countries. when the air humidity is high. This condition is known as ‘putty fish’ (4). it tends to lead to a slight yellowing and softening of the product. and for a heavy salting a ratio of 1:3 salt:fish is used (1). and salt can speed this oxidation process. The fish is then stacked in piles of about 2 metres and the water is allowed to drain away. having a short section at the end of the tail. but the concentration of copper and iron must be kept under 0. Also.3. the fish is restacked with fresh salt after about a week.15 . See Figure 5. addition of levels of 0. producing characteristic pink patches. respectively. respectively (1). Usually it is followed by other preservation techniques such as drying and/or smoking. It takes about 3 weeks until proper salt penetration and moisture loss are achieved. can be dried further to produce ‘klipfish’ or ‘bacalão’. and the salt has penetrated throughout the fish (3). calcium and magnesium ions of impure salts can form a barrier to the passage of sodium ions from the surface to the bulk of the fish. ‘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. when rock salt is used the ‘dun’ patches caused by moulds are prevalent (1). The calcium and magnesium impurities. the fish is mixed with one part of salt per eight parts fish. which consequently inhibits the diffusion of the water from the bulk of the product to the surface. 5. The salt used should be of food-grade quality. It is common for the dried salted cod in the Mediterranean countries to be made using solar salts.1.15% calcium and magnesium. This is because fatty fish turn rancid quickly. 97 . SMOKED AND DRIED FISH 5. mainly in tropical conditions. if very pure sodium chloride is used for curing purposes.1. are likely to cause hydroscopicity of the product and. When pure salt is used. low in calcium and magnesium. However. On the other hand. and essentially free from iron and copper (3). ‘Green cured’ split cod with about 60% of the backbone removed. Salting can be done in a number of different ways.
Fig. although toxin production would be unlikely (4). it appears that water activities much less than 0.1. eviscerated and/or split). such as Staphylococcus aureus could survive and grow at such water activities in the product.1. Food-poisoning organisms. so that the risk of spoilage is high.2 Wet salting or brining As referred to above. The greatest barriers to salt penetration are the skin and scales. when a light brine is used.FISH AND SEAFOOD Dry salting can also be used for small species without the need for opening the fish flat.3.90 are rarely achieved. such as 98 . Only small fish can be salted whole. the process is mainly for flavouring purposes prior to the use of other preservation processes. For large fish it is advisable to split it before salting. Processing of dried salted cod 5.5. Where fish are salted in their whole form. and the uptake of salt is dependent on the type of fish (fatty or lean) as well as its presentation (whole.
44 2.47 6.41 4.41 6.04 5.06 4.85 3. but as it is placed in containers.5 4.23 3. and a red burgundy colour will develop in the product. The main objective of the brining method of salting fish is to limit air penetration during processing.98 5. The fish are kept immersed in their own brine near saturation point for several days. headed and gutted.88 5.78 6.23 2.09 8.61 3.80 5. the fish enzymes play an important role by maturing or ripening the product.30 2.70 6.76 4.90 (1).45 3.11 4.98 6.26 4. sardines.19 3.11 3. Anchovies are the main fish used for these purposes.54 5.26 7. 5.3 days.3 Dry / wet salting In this process.70 3. thus. TABLE 5. the water that comes out of the fish forms a brine in which the fish stays immersed for 2 .11 9.66 5.II. For preservation purposes.5 3. with a shelf life of about 4 weeks (water activity (aw) about 0. the liquid that comes out of the fish forms a brine.78) (1).76 4. For example.76 4.91 2.51 3.95 3. This is why it is the preferred method for fatty fish. See Table 5.86 2.65 2.33 5. and can be collected and replaced again over the fish.96 1.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. limiting the amount of rancidity that can develop.5 10.09 8.76 4. It is then taken out and dried in the sun or in kilns until about 35% moisture in the product is obtained. Fresh sardines. the product obtained is called gaspé-cured (5).98 6.60 2.85 3. can be cured in a saturated brine solution for 6 days and.99 2.26 3.76 99 .0 3.35 4. if split fish is dry-salted.51 5.41 3.74 9. Here.16 7.25 5.00 3. herrings and other fatty fish can be used.17 5. Histamine formation is of importance when using scombroid and some clupeidean species.84 1.23 2.26 1. The brining step for smoking purposes also helps the product to develop a desirable glossy appearance. SMOKED AND DRIED FISH smoking.5 3.1. but mackerel.03 2.03 4.73 1.0 4.56 6.17 4.05 7.10 1.41 5.22 6. the brine is then replaced over the fish. Histamineforming bacteria do not generally grow at water activities below 0. There are several variations of this process. after being drained and pressed at 465 kg/m2 for 15 hours.46 4. the fish is dry-salted.26 2. they can be stored at 28 °C. a saturated brine is used.0 2.CURED.61 3.66 5.23 8.64 4.44 2.25 5. with pressure being applied to the top of the container to help the brine to drain away.3.76 4.78 2.49 4.
but it has further smoking. whereas the latter method preserves the fish for only a few days.3 Marinating / pickling Fish can be brined in solutions based on organic acid instead of salt. In a second step.FISH AND SEAFOOD 5. The fish can be soaked in a high concentration of vinegar (about 4 . ‘ikan kaju’. or a pH <5 and they are stabilised for a limited period. usually flavoured with onions and hot chillies. Herring can also be cured for several days in an 80 to 90º brine with 2. flavouring (sugar.4% of salt) for distribution and sale (1). All these products are ‘semi-preserves’.6 days without refrigeration (1). However. Ceviche. Another salt-boiled Indonesian product. The first process can give a longer shelf life. giving a cooked appearance to the product.2 Salt-boiling Fish can be boiled after being dry-salted in a watertight container and heated for at least 1 hour. The cooked portions are poured into bowls containing gelatin dissolved in water (10% solution) and sometimes a small amount of vinegar. is made of different varieties of fish. Once cooled. they are packed for consumption within 2 weeks at chill temperature (6).2% acetic acid and 2 .30 minutes.5% 120grain distilled vinegar. a Peruvian product.3. placed in a marinade of lime juice. A much firmer product is obtained when the fish is salted prior to acidification. then packed in barrels with a 70º brine. The product is usually consumed raw with the high strength acid solution softening the bones enough for them to be eaten. The Indonesian ‘pindang’ is made by boiling the fish in a saturated salt solution and it is sold as is. which is dipped into a boiling brine for 10 . or can be boiled by being placed in a basket. The lime juice plays the role of the vinegar. and lasts for 3 . drying and salt-boiling steps until a hard translucent block is obtained (4). even if the product is hermetically sealed.2 months. As referred to above. outgrowth of bacterial spores after processing may occur (4). and spices) is added to the brine. Afterwards. and packed (3). the occasional implication of pindang in cases of foodborne illness. There is epidemiological evidence that these products have been the cause of some 100 . underlines the need for careful control (4). until the flesh is soft. containing 1 kg of salt for every 25 kg of fish. usually requiring refrigeration below 10 °C for longer-term storage. and even death. 5. up to 1 . and 6% salt. has a shelf life of several years. After this. They will have a salt content >6% (w/w) in the water phase. only the nonspore-forming organisms are eliminated in the process and. it can be repacked in milder brine (1 . they can be cut and cured for 3 days in a solution with 3% white distilled vinegar. Traditional jellied eels are a product where pieces of fish are cooked in boiling water. Usually. Other preservation techniques can be used for extending the shelf life of these products. Another variation uses sour cream instead of the second brine step. salt. because of the salt content of the product.3.7%) for as long as 3 weeks. herring or other fatty species are used.
The most common method used for fish products is the air-drying process. marinated products will keep for several months at a temperature of 4 °C. under a light pressure. Italy and Spain (5). the fish is dried naturally in a relatively cold environment. It takes about 2 days to marinate completely. In Scandinavian countries. used as an appetiser. Shark pectoral fins. Fresh dill is placed between the layers of fish. Some bacteria and enzymes are still active in the presence of acid and salt. jellyfish. In the first case. split and dried gadoid species) are traditionally made in cold climates. and the stacks are placed in the refrigerator. bacterial toxins and parasites (7). In some coastal areas of Portugal. which are placed in a mixture of salt. Stockfish products have a golden brown colour and are very hard.CURED. such as Listeria monocytogenes and Clostridium botulinum. They are an important dried product. It is important to assure the quality of the raw material. air flows naturally. it being preferable to use frozen salmon or to freeze the product at the end of the process to avoid parasite survival. are generally air-dried at sea. are sundried and.) where no acid is added to the process. after frying. Matjes are made of fat young herring. South America and the Caribbean. sea cucumbers and seaweeds. and the flesh will break down. then mild-cured with special seasonings and packed in barrels with 80 º brine (5).There are several products named marinades (matjes. Stockfish is extensively used in Portugal.4 Air drying This is probably the oldest method of fish preservation. but sometimes it is dried without prior salting.5. sugar and spices. the fish is dried after salting or brining. by using either solar heaters or mechanical dryers. Stockfish (dressed. gutted. Other products. Usually. are likely to survive in these mild-processed products. Dried shrimps are widely used in Asia.3. such as squid. and a fine product is obtained. are commonly dried in Asian countries. some fish species such as moray eel and varieties of dog fish. 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). At pH levels below 4. a very fine delicacy for Oriental food lovers. etc. while in the latter case there is a provision for fans or other means of mechanically moving the air (1). gravad fish. Foodborne pathogens. SMOKED AND DRIED FISH foodborne diseases related to the presence of biotoxins. Gravad fish is a Scandinavian product made of salmon fillets. 5.0) (7). 101 . depending on the thickness. it will last for many years (5). They are not cooked prior to consumption and they have a limited shelf life even at refrigeration temperatures. still made without prior salting on a large scale. Provided the product is kept in dry and fairly cool conditions. Dried octopus is also a very common dish in the interior of the country. the temperature and the amount of pressure applied (5). mixed with salt. These can be considered as ‘lightly preserved’ products with a salt content <6% sodium chloride (w/w) in the water phase and low acid content (pH >5.
FISH AND SEAFOOD When the fish is sun-dried in tropical conditions. and it is exported to Portugal. It is important to balance the drying rates and quality/safety issues. The amount of moisture transfer from the fish to the air is dependent on the relative humidity . Dried salted products must have a water activity below 0.III. and the water must be changed several times before cooking. They must be soaked in water. Although controls are in place to ensure that products processed in such relatively uncontrolled conditions do not enter the supply chain. by warm air circulating in a drying chamber. botulinum growth might occur in the bulk of the product.g. aureus) or spores (e. the ‘green cured’ cod is rinsed or washed with water and then hung to dry either by the sun and breeze or. In many such artisanal fisheries the drying is simply conducted by spreading the fish on the beach. Spain. if the rate of water removal is too high in comparison with the diffusion rate. Staph. more commonly nowadays. the better the moisture transfer. neither it. Whilst the production of dried fish products is tightly controlled in industrialised countries. However. that may have contaminated the product during the drying process. The fish must be stored in reasonably dry conditions to avoid moisture absorption. France. Whilst the low aw of the final product will inhibit microbial growth. the diffusion of the bulk water to the surface of the fish is blocked. spoilage and possibly C. botulinum). Portuguese and Spanish fisheries cannot satisfy their own countries’ demand for cod. To prevent this. C. 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. nor subsequent cooking operations will affect the presence of toxins (e. the fish must be dried slowly enough to avoid case-hardening. making a water barrier and consequently. Traditional processing conditions provide ample opportunity for product contamination from a wide range of hazards and sources.83 (9)). The product is dried to a final water content of about 25 to 38% (3). When no more water can be removed from the fish. some of which are listed in Table 5.g. Owing to restriction of fishing areas for Spanish and Portuguese vessels since the 1970s. the protein must be set or denatured with low-temperature drying before applying higher temperatures. and increasing amounts of such products are making their way into the market place.75 (1). After dry salting. the skin will form a hard case (case-hardening).the lower the humidity. the quality is more difficult to control. 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. Iceland and Canada. tarpaulin. or wooden drying rack to dry in the sun. which leads to mould growth (aflatoxins. In conclusion. toxic secondary metabolites of the moulds Aspergillus flavus and Aspergillus parasiticus are not produced at aw <0. A highly demanded dried fish product in Mediterranean countries is dried salted cod. the product can be held for months at room temperature. skin up. Italy and South America. but fast enough to avoid spoilage caused by bacterial and enzymic activities (1). 102 . and has the primary objective of creating a product with specific taste and textural characteristics. Thus. typically in stews and soups.
In the mechanical kiln. Top-quality fish must always be used. mussels and many others from tropical countries. are some of the ways of reducing the polycyclic aromatic hydrocarbon (PAH) compounds (10). white mangrove and abura has an inhibitory effect on some microorganisms. eel. herring.CURED. mackerel. as referred to above.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. temperature and humidity can be automatically controlled and the process is faster because the directed air movement draws the smoke through the fish. It can be used for cold or hot smoking.5 Smoking Smoking today. sprats. aureus and Saccharomyces cerevisiae (10). Aroma and flavour are a blend of smoke components (3). However. making full description impossible here. is mainly a process for flavouring fish rather than a preservation technique. such as Escherichia coli. using electrostatic filtration of the smoke. cod. botulinum (spores) Pesticides / Chemicals Physical Glass / sand etc. Proper cleaning and preparation prior to smoking and washing of the fresh or frozen fish by water spray or a continuous 103 . oysters. It has been reported that the smoke from mahogany. Preventing the occurrence of carcinogens during smoking has been an issue for the past several years. cod roes. There are many fish species that are used for smoking purposes . The quality of the smoke is related to the type of wood used. Maintaining the temperatures of pyrolysis between 425 and 200 °C.3. clams. Staph. and the use of smoke generated by superheated steam or liquid smoke. The smoking procedures vary widely. haddock. aureus (toxin) C. SMOKED AND DRIED FISH TABLE 5. tuna. trout. Blends of various woods can be formulated to produce sawdust with specific flavours.salmon. The production of smoke by wood or sawdust burning can be done in a mechanical kiln or in traditional chimney kilns. swordfish. Hardwoods are considered better because of the milder flavour they impart to the food. 5. softwood colours the product quickly.
3. Drying should be minimal at the start. temperatures and times used in processing coldsmoked fish are very favourable for the proliferation of food-spoilage and foodpoisoning microorganisms. wooden boxes or master cartons (10). Some herring products. canned or bottled. are boiled before smoking (12). it will reduce the salty flavour. Sugar. are not cooked prior to consumption. tuna and swordfish. Also. but this concentration will again depend on the type of product and the length of the brining stage. Thus. thus allowing the deposit of smoke on the fish surface. but a ‘safety’ (i. owing to the elution of soluble proteins. Depending on the type of product. the excess salt is removed with water. after being smoked need to be chilled immediately to 4 °C before being packed. thawing must be done slowly for 12 .10% of the weight of the fish. filleted. as well as many other products prepared in the same way. 5. all cold-smoked products should be cooked before consumption. skin off or on. The fish can be dry-salted using a weight of salt 5 . Cod roes. which is important for the appearance of these products (4). however. whether cold. an attractive glossy pellicle on the fish fillet surface is achieved after draining. 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. Sugar can be added together with the salt.e. it will double the drying time in the kiln to get the same total weight loss as for dry-salted fillets (11). 104 . low temperatures will slow the uptake of salt. but it will increase towards the end of the process (10). 70 – 80 º brine is commonly used. the highly desirable cold-smoked salmon. usually dry-salted and cold-smoked.1 Cold-smoking The major difference between cold. smoked cod fillets and finnan haddock are usually cooked before consumption. spices. Thus.and hot-smoking is the temperature of the process. such as smoked salmon-trout. The salting period is dependent on the size of the fish and its presentation (whole. However. If wet salting is chosen. it will take up salt twice as fast as fresh fish (10). If frozen fish is used. Wet salting or brining is preferable to dry-salting when smoking fillets because. shrink packaged. In cold-smoking. the following step is salting. or quick-frozen and stored at -18 °C (2). When frozen/thawed fish is used. pasteurisation) temperature is not achieved in any stage of the process.50 ppm) should always be done just before process initiation (3). they can be vacuum packaged. giving the product the desired appearance and flavour (4). Brining at 10 15 °C is recommended (10).or hot-smoked. as it retains more moisture. gutted or filleted.18 hours at 4 °C. and the cold smoke is denser. All the smoked products. The maintenance of temperatures below 30 °C avoids protein coagulation. covering the fish surface completely (10). The length of the smoking process for such products is much greater than for the hotsmoked fish. Whether the product is prepared headed. temperatures must be kept under 30 °C throughout the process. skin on or off).FISH AND SEAFOOD water flow system with chlorinated water (25 . or packed in retortable foil pouches.5.
can grow with salt levels of 10% in the water phase. A typical temperature set is as follows: firstly. botulinum type E the product must have at least 3% salt in the water phase and be stored below 5 °C. can be isolated from seafood. This temperature should be maintained during storage and distribution (3). thirdly.3. 5.CURED. an initial humidity of 70% seems to be preferable because it is a good compromise between maximum smoke adsorption and minimum casehardening (1). SMOKED AND DRIED FISH During cold-smoking.5. and their sensitivity to salt (13). secondly. as. in order to produce toxin. Smoked salmon fillets are usually vacuum packed whole or after slicing. Neither the smoking temperature nor the salt content is enough to kill the organism. refrigerated storage and vacuum packaging still allow its growth (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. a drying period at 30 °C. The exception is vacuum packed fish. Although other human pathogens that can grow at refrigeration temperature. are of concern when smoking fish from tropical waters (13). monocytogenes. there is no point in the process that can fully assure the absence of L. If the refrigeration chain is maintained. Vacuum packing is recommended only for coldsmoked salmon. if a 105 . To avoid growth of non-proteolytic C. and hotsmoked salmon (kippered) (3). thus allowing any psychrotrophic C. typically mesophilic. which underlines the danger of storage at abuse temperatures of >10 °C (13). Furthermore. they are more sensitive to sodium chloride and unlikely to grow in smoked products (13). On the other hand. Hot-smoked products require a longer cooling stage. These products should be vacuum packed at room temperature. but they should be cooled to a temperature of 10 °C within 3 hours. See Figure 5. Smoked products can be also packed under controlled-atmosphere packaging. such as Yersinia enterocolitica.2. Afterwards. The protein must be set or denaturated with lower temperatures before the application of higher temperatures to avoid case-hardening. and the recommended gas mixtures are 60% carbon dioxide and 40% nitrogen (15). the products are cooled. condensation will occur inside the pack. a partial cooking period at 50 °C. but the proteolytic types A and B. botulinum type E present an even longer time than before to grow and produce toxin (14). Pathogenic vibrios. stored at 4 °C and packed. further cooled to 3 °C within 12 hours and then packed. if they are taken from a chill room at 3 °C and packed. Length of the smoking process at each stage will depend on the species and the type of product. During smoking. if present.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. gravad salmon (also called gravad lax or gravlax). a final cooking period at 80 °C (12). Vacuum packing extends the shelf life of the lightly smoked products by suppressing the growth of other bacteria and moulds.
Process of cold-smoked sliced salmon 106 .FISH AND SEAFOOD product is packed too warm without being cooled to room temperature. Fig. 5.2. it will have a reduced shelf life. and moulds will grow on the product (12).
including ‘umami’ (16.3.3½ hours using the typical temperature set for hot-smoking referred to above (12). They are generally not cooked prior to consumption. Katsuobushi. white fish about 8 days. 23). rewetted and remoulded to give the desired shape and consistency. moulding it into long thick sections. The classical liquid smoke is 107 . hot-smoked fish can be classified as a ‘heat-treated or pasteurised’ product (7). cooking in brine. Hon kare bushi is a form of Katsuobushi from which shavings are cut to make aromatic stocks for use in Japanese dishes (24). then hot smoking and finally sun drying it. 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). resulting in a characteristic flavour (20. and substances unconnected with smoke but with a smoky flavour or smell. When refrigerated at 3 °C. 18). Aspergillus tamarii) and several Penicillium spp. A type of dried hot smoked tuna called masmeen or masmin is produced in the Lakshadweep islands of the Indian Ocean. The production method is similar to masmeen. break down of fats and the moderation of smoke flavour. fatty fish will remain in good condition for 6 days. Hot-smoked products made from white fish will usually keep better than those from fatty fish. Mackerel.6 hours and smoked for 3 . Vacuum packaging retards the onset of rancidity when fatty fish is used. The product is often allowed to go mouldy by incubating it in a chamber containing a natural fungal flora. including Eurotium repens and Eurotium rubrum. Growth of these moulds on the tuna results in the extraction of moisture. They will keep well frozen. SMOKED AND DRIED FISH There are several species that are usually hot-smoked and the process can vary markedly. 22.3 Liquid smoke Smoke-flavoured fish represents a small percentage of the total smoked fish industry output. but mechanical drying is used instead of sun drying. to have no further advantage over other packing methods for other fish (12). is a boiled smoked fish product that is used in Japanese cooking to impart flavour. apart from the improved appearance. are usually 80º brined for 5 . and many closely related Aspergilli (Aspergillus ochraceus. Eels. Masmeen is usually prepared from bonito tuna by eviscerating and filleting the fish. The product is removed from the drying cycle.5.CURED. are usually hot-smoked after being immersed in 80º brine or dry-salted (6). 5. but seems. 21. chilled or freeze-thawed. 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). similar to masmeen. In terms of safety. a very expensive and fashionable smoked product in the Netherlands. synthetic smoke flavours. 17. The composition of commercial liquid smoke can vary widely depending on the method of manufacture. (Penicillum cyclopium and Penicillium puterellii) (19). Boiled or steamed clams. they can be stored for 6 months at -30 °C and for even longer when frozen in vacuum packs (12). Aspergillus ostianus. Aspergillus oryzae. are used. Natural smoke extracts. such as yeast derivatives.
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).FISH AND SEAFOOD smoke condensate that is dissolved in water or oil. spraying is preferable because it is easiest to control flavour and acceptable with this method (3). and less than 0. drying. or applying it as an atomised spray within a modern automatic smoke kiln.8% and still provide protection for 7 days (25).3.6 to 2. 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. Liquid smoke was considered to provide advantages of safety.4 Spoilage Brining. pickling and marinating fish were originally used to lengthen the short shelf life of fresh fish. It has been shown that liquid smoke can reduce the formation of PAHs (3). Whilst neither the CO or the carboxymyoglobin complex are harmful in themselves.06 μg/kg of benzoanthracene in the final foodstuff (28). There are two groups of chemicals of concern in smoke: polycyclic aromatic hydrocarbons (PAHs) and nitrosamines.03 μg/kg of benzopyrene. it has been reported that the liquid smoke-treated product has less pronounced aroma and flavour (25). 5. so as to impart an unnatural red colouration to the flesh.5. applying it as a dip after brining. nutritional neutrality and antioxidant activity (27). botulinum types A and E spores in several fish species that were stored at 25 °C for 7 and 14 days (25). When liquid smoke was used. 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. In experiments comparing traditionally smoked fish with liquid smoke. injecting it into the fish flesh. established in its guidelines that smoke flavourings added to the food product must assure a level lower than 0. or smoke extracts in organic solvents.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. consistency of flavour and colour. the salt level could effectively be reduced from 4. smoking. The process relies on the generation of the smoke to produce elevated levels of carbon monoxide (CO) which interact with haemoglobin in the muscle to form the very stable bright pink/red carboxymyoglobin complex. 5. Fresh fish spoilage may be measured by 108 . The European Scientific Commitee for Food. both of which are considered potential carcinogens. Smoke-flavoured fish processed using liquid smoke can be prepared by including liquid smoke in the brine. According to some recent research.
the pickle and dry-salting preservation processes require further processing for significant shelf life increases (e. which may also be produced to describe the quality attributes and their loss over time for processed fish. marination or drying). but. and. When acid marinating or smoking. Spoilage needs to be carefully defined for each product. these halophilic organisms may grow and cause spoilage. Stronger and longer brining has an effect like that of pickling.. Often. the shelf life may be estimated and investigation made into the causes of the loss of sensory quality. This is not usually the case for smoked fish as the residual salt concentration is not high enough to inhibit all the non-halophiles. a very dry product is required to produce 'shelf-stability'. Most marinated and smoked fish products are first brined or salt-pickled. contact with the bacteria 109 .3%). also. For some products. a process similar in microbiological effect to that of saturated brining. or extended refrigerated storage. The microflora attached to the fish is relatively unaffected by light brines used before smoking.CURED. Even saturated brines allow some halophilic organisms to grow. Within the limits of safety. Usually. otherwise. limits the types of microorganism capable of growing. the fish flora are transferred to the brine mix. and coryneform bacteria. In salts.000 cfu/ml of brine may build up. the bulk of the microflora comprises Bacillus spp. solar salts investigated carried the highest load and included halophilic organisms as a result of their production method (open-air evaporation of sea water). and high concentrations are usually required to inhibit their growth fully. however. the fish are first cleaned (known as ‘rousing’) by a salt rub to remove any outer slime. rock and manufactured salts did not usually contain halophilic bacteria (29). Upon salt packing. chemical oxidation. Endogenous marine fish bacteria prefer to grow in the presence of salt (marine concentration approximately 2. these bacteria tend to continue to dominate the flora in the liquid. and of smoke to add chemical preservatives. contaminant bacteria from other materials used to produce the product.. pickling and dry-salting all introduce bacteria to the fish. therefore. Pickling and/or the anchovy process uses solid salt extraction of fish fluids to form a brine solution (cure). Some brines are re-used over a period of days and a load of 10. the fish are cut and splayed to allow greater surface contact with the salt. Salts used for brining have been found to carry a microbial load of between 10 and 1000 organisms per gram of wet salt. spoilage is due to endogenous microorganisms and enzymes of the fish. SMOKED AND DRIED FISH grading schemes or quality indices. both of these will affect the spoilage patterns. Brining. The use of drying and/or salt to lower the water activity. Once placed into brines. the production process will modify the microbial flora present on the product. for others. as the fish are added.g. which are found in the salt. Once these changes are defined. When heavy brining or salting is important for product shelf life. a product may become undesirable for consumption as a result of changes in its sensory characteristics. for some. mainly comprising corynebacteria and Gram-negative bacteria from the fish and water (29). or their products require refrigerated storage. the consequent reduction in the pH will help to inhibit spoilage organisms. Micrococcus spp.
rancidity due to lipolysis. the best way of controlling the organisms is to refrigerate the fish. protein breakdown. sometimes covering the whole of the fish with a whitish. (syn. including A. ‘Pink’ describes a condition associated with surface growth of the pigmented halophilic bacteria Halococcus spp. The bacteria form coloured colonies on the surface of salted fish prior to drying. and Halobacterium spp. pisce was the main fungus isolated. Several Penicillium spp. W. repens and E. together with a range of Aspergilli.. or fat breakdown) caused by enzymes rather than by microbial growth. 33). if not too contaminated. ‘Sliming’ or ‘putty’ fish is a condition that occurs in poorly or insufficiently salted cod (e. halophilica was isolated 110 .FISH AND SEAFOOD in the salt is made and the flora will develop accordingly in the blood pickle. it was found in about 50% of samples examined. or after drying if the fish is stored in excessive humidity. yeasts. Their growth produces noticeable spoilage of the fish appearance but is also controlled by refrigeration. A. and still produce a reasonable dried product. ‘dun’ is not characterised by the production of offensive odours. At the pre-drying stage. powdery growth. Therefore. may also be found in the salt used in production. spoilage of these pickled and dried fish in refrigerated storage tends to be due to chemical changes (textural changes due to proteolysis. most notably Penicillium citrinum and Penicillium thomii. A number of spoilage organisms can grow and produce conditions such as ‘sliming’. The microbiology of heavily salted products is dominated by Gram-positive. Unlike ‘pink’. This condition is often associated with case-hardening. niger. 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). Aspergillus sydowii. as the water activity of saturated brine solutions is low enough to inhibit most bacteria. Basipetospora halophila. E. The bacteria are osmosensitive and will lyse in water. although it may occur in other salted fish. halotolerant or halophilic micrococci. Eurotium amstelodami. Insufficient salt penetration into the flesh leaves areas where Gram-negative rod-shaped bacteria and micrococci grow and cause a putrefactive spoilage. spore-forming bacteria. 32. like the ‘pink’ organisms. sebi is considered to be the main spoilage agent of dried fish produced in temperate regions (30). including hydrogen sulphide (26). though only the latter two species were considered likely to have grown on the fish (32). Studies conducted on dried and salted fish from Indonesia (31. flavus. Aspergillus wentii. yeasts and fungi. and Aspergillus penicilloides. as little if any growth will occur below 5 °C on salted fish (29). gaspé).g. (syn Oospora halophila) and Polypaecilum pisce are two white halophilic moulds that are associated with spoilage of salted fish (31. were also isolated. rubrum are also common isolates from dried fish. ‘pink’ and ‘dun’. Bacteria from these two genera also grow sufficiently in brine and even salt to cause pink discolouration. Only halophilic microbes may grow. B. The condition describes the growth of the halophilic moulds (Wallemia sebi. The metabolism of the organisms will produce off-odours. 34) have shown that P. Sporendonema sebi)). it is possible to wash off the infected parts. which. lactic acid bacteria (LAB) and moulds.
so the bacteria spoiling these products will be psychrotrophic or psychrophilic. Vibrionaceae and Enterobacteriaceae may all play some part in shelf life limitation. 111 .CURED. including A. around the water activity of saturated sodium chloride. botulinum type E.V gives details of pH ranges and aw minima for the growth of some important spoilage microorganisms and pathogens (38). The preservative effect of the smoke will inhibit some spoilage microorganisms. but this will not prevent the growth of spoilage bacteria. has been isolated from dried fish (35). Marine vibrios were the dominant flora in smoked salmon that was drysalted. The brining to 3 . It has been reported that the type of salting (dry.5% sodium chloride in the water phase will inhibit some bacteria. 36). Technical studies demonstrate the ability of many of the xerophiles above to grow at typical water activities seen in dried salted fish (32. most marine bacteria are moderately halophilic and are not effectively inhibited by these low salt concentrations.or wet-salting) in the cold-smoked salmon process leads to different microflora growth at the onset of spoilage. The salt in the water phase must be high enough to inhibit the growth of C. flavus. The heat during hot-smoking will kill vegetative microorganisms but not all spores. In Vietnam. LAB. or any smoking process where temperatures in the interior of the fish do not reach >50 °C for a significant period of time. For more information on spoilage see Table 5. Cold-smoked fish will contain microbial flora representative of that of the raw fish. Reports for vacuum packed cold-smoked salmon have identified more than one organism associated with spoilage. with a lower aw limit for growth of 0. Vacuum packing limits chemical oxidation. but often colour changes due to chemical oxidation of the smoked product will limit the shelf life.IV. so the most probable spoilage organisms will be spore-formers or post-smoking contaminants. Aspergillus clavatus and Aspergillus niger. will have a different residual post-smoking flora from 'hot-smoked' fish. and microbial spoilage is more likely. but. SMOKED AND DRIED FISH infrequently at low levels.747. Fish processed by cold-smoking. Smoked fish products tend to have a light brine treatment. Pseudomonadaceae. however. which does not create the salty conditions required for spoilage by those halophilic microorganisms. able to grow on dried salted fish. where a high internal temperature is reached. Refrigeration of hot-smoked fish will greatly reduce the number of organisms able to grow and will lengthen the shelf life. 33. Refrigeration is necessary. Table 5. it was. not the growth of spoilage organisms. whereas a mixture of Enterobacteriaceae and LAB were prevalent in brineinjected salmon (37). as already stated. Aspergillus spp..
FISH AND SEAFOOD TABLE 5. Matjes v3 + ++ v - +++ ++ ++ - v +/- ++ ++ ++/++/- Pindang Bacalao + +++ Gaspé cure Anchovie ++ Anchovies ++ - - - ++ ++ Saturated brined fish Salted then boiled Salted. LAB. Enterobacteria and PPC Spore formers and PPC Variable Salt marinades Salted then boiled Brined then boiled Salted then dried Canned Fish Gravad. . + to +++ strength of preservative effect 4 Post-process contaminants 112 .IV Spoilage of salted.variable. sebi and B. halophila Halophilic spoilage organisms Halophilic spoilage organisms. Dun. smoked and dried Citrus marinade Dried Brined or salted then liquid or cold smoked Brined or salted then liquid or hot smoked 1 2 +++ Jellied eels + Ikan kaju Ceviche Stockfish Coldsmoked fish Hotsmoked fish +++ +++ ++ +/++ + + ++ ++ - ++ + + + ++ ++ ++ ++ + ++ + ++ Antimicrobial compounds added Refrigeration applied 3 v. Histamine producers Halophilic spoilage organisms Spore formers and PPC None Acid-tolerant spoilage organisms Xerophilic moulds Fresh fish spoilage organisms. and Halobacterium spp. boiled.W.not used/unchanged. 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.
It has been shown that. with this change.0-9.8 1. salted and pickled products accounted for 10.86 0. SMOKED AND DRIED FISH TABLE 5. when present in the raw material. aureus Yeasts Moulds Lactic acid bacteria pH 5. sodium 113 . pressure on the traditional methods of curing. and C. botulinum type E and production of toxin. In the United States. botulinum. respectively. types A and B are not affected by 3 .5 5.V pH ranges and aw minima for growth of microorganisms (38) Organisms Most Pseudomonadaceae C. Since neither the smoking nor the drying parts of this curing process are particularly severe. but the concentration of salt in smoked salmon or trout (usually varying from 1 4% salt) is not high enough to prevent growth.9 0. popular demand for healthy foods has led to a change in the tastes preferred by the public and.5-10.5% to 5%. and as botulism is a serious hazard in foods.61 Organisms Most Pseudomonadaceae C. temperature ≤5 °C and salt ≥3% in the water phase is sufficient to prevent any growth of type E. botulinum type E Staph.5 4. but if the fish products are stored at temperatures <10 °C. the non-proteolytic type E strains can grow in up to 5% sodium chloride. 3% sodium chloride in the water phase is enough to inhibit growth of type E for at least 30 days (39). However. 9 and 8 of the outbreaks. smoked and dried. Before.CURED. although approximately 10% salt in the water phase is necessary to fully inhibit the mesophilic types A and B strains (14). aureus Most spoilage bacteria Most spoilage moulds Halophilic bacteria Xerophilic moulds Aw 0. botulinum Staph.5% of salt in the water phase and great care must be taken to avoid any temperatures >10 °C (13). botulinum type E. although the combination of the two factors. the importance of cold-chain integrity must be emphasised.8 0.5-10 4. The concentration required to prevent growth at room temperature can vary from as low as 3. A number of surveys have shown that fish from temperate waters is mainly contaminated with the psychrotrophic C. but the presence of the mesophilic types A and B is of concern in fish from warmer waters (13).97 0.1 Pathogens: Growth and Survival Clostridium botulinum In the last few decades.5 5.5-8. the fermented products being the most dangerous group (7).5-8. under optimal conditions.5. could not grow and produce toxin.75 0.5 0-11 3. the products were heavily salted. Examination of 165 outbreaks of botulism caused by fish products showed that smoked. Sodium nitrite in conjunction with sodium chloride is known for its ability to inhibit C. The presence of salt in cured products has a great effect on the growth of the bacteria.97 0.
5. the human nose. they can survive lightly cured processes when the sodium chloride content is less than 5% and water activities are higher than 0. the risk of infection can be eliminated by proper cooking.5. Once the product is contaminated. more importantly. The degree of concern in coldsmoked salmon is low unless temperature abuse >10 °C occurs (13). they will grow rapidly (7). related to the consumption of a raw smoked fish meal.2 Staphylococcus aureus This is a ubiquitous organism but its main reservoir and habitat is animals and. gills and guts as well as in several lightly preserved fish 114 . The FDA established that hot. but. aureus growth.94 (7). Thus.86 and 10% sodium chloride. but requires higher temperatures for toxin production (>10 °C). 5. if the person handling the product is a carrier of Staph.5% salt in the water phase. It is a halotolerant bacterium able to grow at water activities as low as 0. An outbreak of Shigella in an Italian cruise ship. the organism may contaminate the product. after processing. Infection by Shigella sonnei has increased in the UK. Shigella and E. The human carrier rate can be as high as 60% of healthy individuals with an average of 25 . the organism can survive most of the light curing processes.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.5. if storage temperatures are over 10 °C. it should contain not less than 3. if it contains sodium nitrite ≥100 ppm. they do not grow well in the raw material. has been reported (42). As they are poor competitors.3 Enterobacteriaceae The pathogens Salmonella. are particularly susceptible to contamination and.0 . 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. aureus. As with other organisms. However.5. 5.5. growth and toxin formation may occur. if recontamination occurs and storage conditions are favourable. its occurrence in a final product is due to contamination during processing by an infected asymptomatic carrier with poor personal hygiene. monocytogenes is a ubiquitous organism that has been detected in fresh fish skin.30% of the population positive for enterotoxin-producing strains (41). Sliced smoked fish products. Usually. coli all occur on fish products as a result of contamination from animal/human reservoirs.or cold-smoked fish should have at least 3.4 Listeria monocytogenes The psychrotrophic L. and the minimum pH for growth is 4. as they require more handling. In foodstuffs. when the bacterial load of the product is lower.0% salt in the water phase (30). throat and skin. The organism is mesophilic with a minimum growth temperature of 7 °C.4. but.
the round worm Anisakis spp.95 (45). a number of fish products. Vacuum packaging (VP) of sliced smoked fish creates a microaerophilic atmosphere. Thus. SMOKED AND DRIED FISH products such as cold-smoked fish.92 . As freezing to -20 °C and maintaining this temperature for at least 24 h will kill all the nematodes. which means that cooking a fillet 3 cm thick for 10 min at 60 °C will kill any worms present (48). from freshwater fish. 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. Marine LAB is part of the natural flora of chill-stored vacuum packed salmon and cod. have been shown to have an anti-listerial activity in many food products. such as ‘lightly preserved’ fish products. As they are heat-sensitive. favours growth of psychrotrophic LAB. marinades (ceviche) (43) and gravad fish (44). salt levels of 8 . are considered unsafe. pH 4. and the tape worm Diphyllobothrium spp. do not survive 1 min at 60 °C. in combination with curing agents and refrigerated storage. widely used as starter cultures in food processes. when an outbreak of nine cases of listeriosis was suspected to have been caused by gravad or cold-smoked trout (47). and other local traditional products (7). The use of LAB in fish preservation is a topic that has been studied in projects (UP.6 and water activities below 0. The round worms. the USA established ‘zero tolerance’ for the organism. gravad fish. cold-smoked fish. which. Until 1997. Of microbiological concern in the northern hemisphere are two parasites . which means that the process must conform to a level of ‘no detectable L. which include intermediate hosts. which considered L. most of them are of little concern to public health. has also been detected in cod (7). from seawater fish. a short period of freezing either of the raw material or of 115 . As its presence has been reported in lightly preserved fish products whose processing chain cannot fully assure the absence of the organism. Its limiting conditions for growth are 0 . and in CT95-1207FAIR). 5. both being detected in cold-smoked salmon. Two outbreaks in the United States have been reported to be linked to salmon (10).5 Parasites Despite the presence of parasites in fish being very common. monocytogenes in levels at or above 102 per gram (46). in the EU Fisheries R&D-FLAIR program. lightly salted caviar. the CT 3162-FAIR program. there were no reported outbreaks related to cured fish. a round worm. Some LAB.0.8 . monocytogenes pathogenic for only specific segments at risk.2.514.CURED. Unlike some European countries. or nematodes. They have complicated life cycles.45 °C. This includes matjes herring.5.9. researchers are looking for other ways of preventing the pathogen’s occurrence or growth. monocytogenes’ in 25 g of the finished product. Pseudoterranova dicipiens. the only concern to public health is when eating raw or uncooked fish products.12%.
<100 >104 >100 N/A N/A <20 20 . Acceptability is based on appearance.<104 100 . and unpasteurised yoghurt.<100 200 .<107 ≥107 N/A <100 <20 100 . soft cheese. monocytogenes) (cfu/g unless stated) <106 106 .<104 20 . monocytogenes Staph. aureus C. cholerae Vibrio parahaemolyticus L. 5. taramasalata and cooked shellfish sampled at the point of sale (adapted from (11)) Criterion Satisfactory Borderline limit of acceptability (cfu/g unless stated) Unsatisfactory Unacceptable/ potential hazard* (cfu/g unless stated) (cfu/g unless stated) Aerobic plate count† (30 °C. salami. and the levels or absence of indicator organisms or pathogens. (not L.<100 20 .<104 present in 25 g ≥103 >100 ≥104 ≥104 <103 103 . † Guidelines for aerobic colony counts may not apply to certain fermented foods. perfringens B. These foods fall into category 5. 48h ± 2h) Indicator organisms‡ Enterobacteriaceae E.<104 104 .<100 10 . the organism should be identified # N/A denotes not applicable 116 . for example.6 Published Microbiological Criteria TABLE 5. ‡ On occasions some strains may be pathogenic. cereus and Bacillus subtilis# <20 <20** <20 <10 20 .FISH AND SEAFOOD the final product should therefore be included in the processing as a means of controlling parasites (7). smell. texture. E.<100 20 . coli (total) Listeria spp.<103 N/A 100 .<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. coli O157 & other VTEC Vibrio.VI Guidelines for the microbiological quality of ready-to-eat smoked fish. If the Bacillus counts exceed 104/g.100 >100 N/A Pathogens Salmonella spp. not detected in 25 g Campylobacter spp.
Regenstein C. Leatherhead. coli (ETEC) L. Assurance of Seafood Quality.H. Moody M. 1994. Marinades. Wareing P. aureus * Vibrio cholerae V. 2007. 3273. 1984. 117 . Doré I. New York. Preservation of fish by curing (drying.E. in The Seafood Industry. 1979. Ed.A. Torry Advisory Note No. monocytogenes Salmonella spp. 3. Urner Barry Publications Inc. Eds. Flick G. Torry Advisory Note No. where 30 ppm is permitted Paralytic shellfish poisoning* Amnesic shellfish poisoning * *These criteria are applied to all fish 5. in Fish Processing Technology. Eds. Fernandes R.F. Fernandes R. Smoked. 9. or presence of toxin 0. cured and dried fish. USA. 6.. 254-82 5. salting and smoking). The Smoked and Cured Seafood Guide.W. Bannerman A. Hall G. botulinum * Limit of tolerance 1x 103 ETEC/g..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. Catching. FAO Fisheries Technical Paper..7 1.8 ppm (saxitoxin equivalent) 20 ppm domoic acid. Handling and Processing Eels. Wareing P. Hot Smoking of Fish. Introduction to Fish Technology. 381-406. 2. 1991. 1990. London. * Staph.. 4. 1982. Torry Advisory Note No.M. 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. Leatherhead Publishing. 1993. Flick G. SMOKED AND DRIED FISH TABLE 5. except in the viscera of dungeness crab. 1997. Food spoilage fungi.M. Van Nostrand Reinhold. References Regenstein J. 7.F. Birnie K. New York.CURED. Martin R. 120-38.. in Micro-Facts: The Working Companion for Food Microbiologists. McLay R. 37(revised). 82 (revised). Van Nostrand Reinhold. 334. 8.. Huss H. Horner W. Horne J. Chapman & Hall. 56. parahaemolyticus Vibrio vulnificus C.
Torry Advisory Note No. Yajima I.. 18. 2007. Effect of dried bonito (Katauobushi) and some of its components on GABA-A receptors. Pelroy G. Shiemori Y.. Biotechnological Applications..E. in Fisheries Processing. Biotechnology. O-methylation of 2. Matsui M... 2324-5. Aoshima H.C. Recommendations for the Preparation of Smoked Salmon. Bioscience. 59 (12). Kanayama T. Martin A. Control of biological hazards in cold smoked salmon production.. 843. 51-81. Oxidation of acetophenone by Aspergillus species and their possible contribution to Katsuobushi flavour.M. Technomic Publishers. Bioscience. Journal of Food Protection.. 118 . 1980.M. Cann D. Lindemann B. Horne J. 6 (6). 14.. Paranjpye R.H. Shuto Y.B. 24. 1994. Peterson M. Degradation and omethylation of phenols among volatile flavour components of dried bonito (Katsuobushi) by Aspergillus species. 17. 1982. Sakakibara H.. Kinoshita Y. 19.. 13.. Ninomiya Y. Doi M. 1989.. Jeppesen V. Asymmetric reduction of acetophenone by Aspergillus species and their possible contribution to Katsuobushi flavor. Biotechnology and Biochemistry. Food Product Design. Ed.. 16. Flavour constituents of dried bonito (Katsuobushi). 1998.K. Chapman & Hall. Martin A. 6 (4). 56 (6). 958-60..W.E. Druschitz A. The discovery of umami. 5 (revised). Hayashi K. 2008.F. 17 (9). Botulism and Fishery Products. Microbiological control for fish smoking operations.. 335-40.C.. Patel T. Perusing Asian soup options. and Biochemistry. 1981. Bannerman A. 22.R. Doi M. Torry Advisory Note No. 45 (10). Fish Drying and Smoking: Production and Quality. 553-72. What’s in a name? Are MSG and umami the same? Chemical Senses. Hosokawa M.. 1995.. Cann D. Mitou M. Ogiwara Y. 11. Dillon R.A. Processing Cod Roes. Doe P. Ninomiya M. 22 (revised). Kinoshita Y. 1995. Huss H.. 935-41. 1977. 18. Matsui M. 1990.. 20... 108 (3). 53 (4). Lancaster..FISH AND SEAFOOD 10.. 2002. Doi M. Shuto Y. Doi M. Inhibition of Clostridium botulinum types A and E toxin production by liquid smoke and NaCl in hot-process smoke-flavoured fish. Food Control.M. London. Eklund M. Agricultural and Biological Chemistry. Bioscience. 6-dimethoxy-4-methylphenol by Aspergillus glaucus and their possible contribution to Katsuobushi flavour.. Packing Fish in a Modified Atmosphere. Bannerman A. 88. Yamauchi H. 21. 1051-5. Biotechnology and Biochemistry. Embarek P. 61 (8). Halpern B. Food Reviews International. Torry Advisory Note No. 27 (9). 840-6. Torry Advisory Note No. 1386-7. Yokayama S. 23. 1984. 15. 1992. 12.P... Food Chemistry. Shuto Y. 1997. 508. Teeny F.. 25.
Cox J. Fungi and Food Spoilage.F. Sirvas S. 98100. Effects of sodium chloride and relative humidity on growth and sporulation of moulds isolated from cured fish. 30. 29 (2). Simini B. 49 (1). 33. Anggawati A... Smoked and smoke-flavoured fishery products.D. V. 742. Hocking A. 348. Borgstrøm G. 1941. Fungal flora of South Vietnamese fish and rice. Washington DC. 14. 1986. Maga J. Hess E. 22. 38.E.E. Boca Raton. An outbreak of Shigella. New York. Seafood Safety. Food Microbiology. 164-9. 1994. 1997. Food Technology. 1996. Hocking A.T. 70-2. Loessner M. Letters in Applied Microbiology. 88-90. Pitt J. 197-208. 38 (3). Studies on Sporendonema epizoum from ‘dun’ salt fish. 74 (2). 2005. 31.J. New species of fungi from Indonesian dried fish. Pitt J. 3 (4). Food and Drug Administration. Smoke flavourings. Shewan J. Oxford. Wheeler K. 36. Hocking A. 107-11.. Sprouse R. Academic Press. The control of botulism in hot-smoked trout and mackerel. Mycotaxon. 29. Nahrung. 32.D. CRC. Incidence of Listeria monocytogenes in an acidified product. Pitt J. Diyaolu S.D. Jay J. Food and Drug Administration..C.F. Lancet. Scientific Committee for Food. SMOKED AND DRIED FISH 26. Hansen L.. Huss H.V.O. US Food and Drug Administration. Journal of the Fisheries Research Board of Canada. Blackie. Hocking A. 12. 34. Frank M. Ahmed F.. 74. 1995. 39. Modern Food Microbiology. in Fish as Food. 27686. Journal of Food Technology. 1971.. The microbiology of sea-water fish.. 35. 351-7.. 218-25. Committee on Evaluation of the Safety of Fishery Products.. 1993. Luxembourg. 311-7. 28...S.R. National Academic Press. 41. Journal of Tropical Medicine and Hygiene. CEC. Røntved S. Tour highlights production and uses of smoke-based flavours. Interactions among xerophilic fungi associated with dried salted fish.M.H. 74.. Studies on salt fish.CURED. in Smoke in Food Processing. Taylor L. Golden D. 40.. 43.I. 5. 1991. 1996.A. Lucas F. Fungi associated with Indonesian dried fish. 1979.. Gill T.Y. Adebajo L. 123-9. 1985. Springer Science. Press Inc. Importance of autolysis and microbiological activity on quality of cold-smoked salmon. 1995. Fuchs R. Ed. Antimicrobial properties of smoke. Commission of the European Communities.D.A.. 119 . Food Research International.A. Pszczola D.M. in Fish and Fishery Products Hazards and Controls Guide. Ed. Cann D. Wheeler K. 37. London.I. 1961. Institute of Medicine. Ed. ceviche. Ed. in Reports of the Scientific Committee for Food (34th Series).B.. Townsend J.I. Journal of Applied Bacteriology..M.K. Rockville.A. 181-8. 1994. 42.. 27.1988. 1991.
1994. Wooten R. 1996. US FDA Centre for Food Safety & Applied Nutrition. Harrison M.A. Food Control.J. 22.. Microbiological guidelines for some ready-to-eat foods sampled at the point of sale. The effect of NaCl on the upper temperature limit for growth and enterotoxin synthesis by Staphylococcus aureus.H... Appendix 5: FDA and EPA Guidance Levels.. Little C. Hurst A. Pelroy G. Loncarevic S. Richards J. Fletcher R. Cured. Incidence and growth of psychrotrophic Clostridium botulinum in food. Bolton F.. 135-85. Peterson M.. 5-7. Clostridium botulinum and its importance in fishery products. Moral A. Hughes A. and smoked fish establishments good manufacturing practices.A. 46. Tham W. Pelaez C.. Huang Y. 188 (3).V. 507-10. Journal of Clinical Microbiology. Inhibition of Clostridium botulinum types A and E toxin formation by sodium nitrite and sodium chloride in hot-process (smoked) salmon.. Journal of Food Protection. Nye K.. Jacobsen M. Round Worms in Fish. Loncarevic S.. Liston J. in smoked and "gravad" fish. 80. Advances in Food Research. Eds. 2001...E. 163-7. 26 (4).FISH AND SEAFOOD 44. http://www. 37 (1). Hobbs G. Davies A.... Journal of Food Protection. Danielsson-Tham M-L. 120 . 1980.. Elsevier Science Publishers B. Canadian Journal of Microbiology. Washington D. Torry Advisory Note No. Garren D. 45 (9). 1980.W.. Communicable Disease and Public Health. Susuki E.N.. Pathogenic microorganisms of importance in seafood. 48. Mentzing L-O. Ribeiro C. Persson I. de Louvois J. Beltran A..8 Further Reading Gibson D.J.R. in Quality Assurance in the Fish Industry. Unnerstad H. Tham W.M. 1992. 35 (11) 2904-7. Danielsson-Tham M. Keeping quality of vacuum packed smoked sardine fillets: microbiological aspects.fda. Paranjpye R. Growth and production of toxin of Clostridium botulinum type E in rainbow trout under various storage conditions.. Ed..C.html Gilbert R.. salted. 197-209. 1991. Eklow A.C. 58 (8). 232-6..W. Acta Veterinaria Scandinavia.A. 13-8. 2000. 45.D.S. Zeitschrift für Lebensmittel-Untersuchung und –Forschung. 1989. Gibbs P.. Roberts D. Prevalence of Listeria monocytogenes and other Listeria spp. 5.. 49-59. 5 (1). 55 (4). 1995. Huss H...cfsan. Eklund M. 833-41. Amsterdam. Donovan T. US FDA Centre for Food Safety & Applied Nutrition.L. Cann D. 863-6. 1982. 3. Ericsson H. Journal of the Association of Food and Drug Officials.gov/~comm/haccp4x5. Food and Drug Administration. Association of Food and Drug Officials...M. 1997.. An outbreak of listeriosis suspected to have been caused by rainbow trout. in Fish and Fishery Products Hazards and Controls Guide. 47. 1976.
58 (5). Dillon R. 1994. 1992. Presence. van Spreckens K. Journal of Food Protection. Occurrence of Listeria monocytogenes in freshwater farms and fishsmoking plants. Poindexter C.. International Journal of Food Microbiology. 844-6.T..K... 1734. Journal of Food Protection. Liston J. Dallas H. 1988. 19-27. 1995. solute. 502-8. detection and growth of Listeria monocytogenes in seafoods: a review. 121 .. Poysky F. Nicolaides L. Eds. in Seafood Quality Determination. Contamination pattern of Listeria monocytogenes and other Listeria spp. Food Microbiology. Competition of food bacteria with Listeria monocytogenes during enrichment culture. 4389-95. Combined effects of water activity. Taylor S. 1987. Letters in Applied Microbiology. Rørvik L. Fate of Aeromonas and Yersinia on modified-atmosphere packaged (MAP) cod and trout. Incidence and sources of Listeria monocytogenes in cold-smoked fishery products and processing plants. Interacting effects of pH. Journal of Food Safety.S. 1991. Slade A. SMOKED AND DRIED FISH Sutherland J. Hitchins A. Beuchat L. 27 (6).178-80.E. 19.. pH and sodium chloride. 1075-80. International Journal of Food Microbiology. 394-6.CURED. 42. 25 (5).. Letters in Applied Microbiology. Davies A. 21 (3). Kramer D... 1994. Keusch A. 11. Jemmi T. 61 (12).. 293301. 1995. Eklund M. in a salmon slaughterhouse and smoked salmon processing plant.W.R. Applied Microbiology. Yndestad M. Pelroy G. 309-18. 217-36.A. Prevalence of Listeria in smoked fish. 866-70. New York. 1973.R.. 309-16. Paranjpye R. Lashbrook L. 1995. 14. 25. and temperature on the growth of Vibrio parahaemolyticus. Elsevier. Letters in Applied Microbiology. Marine toxins of microbial origin.A. Caugant D..C. 1994.... Ratnam S. Predictive modelling of growth of Staphylococcus aureus: the effects of temperature. 11. Histamine production by psychrophilic flora. Roberts T. 23 (1). Applied Microbiology. International Journal of Food Microbiology. temperature.N.. 94-8. Incidence of Listeria in hot and cold-smoked fish. Ben Embarek P. Food Technology. Hudson J. 55 (11).P. Efficacy of high sodium chloride concentrations for the destruction of Listeria monocytogenes.J. and salt concentration on growth and survival of Vibrio parahaemolyticus. 1994. Beuchat L. Peterson M. Patel T.R.. Tran T. 1974. Fuchs R..A. 1992. Bayliss A.
lactic fermentation can play a significant role. Carbohydrates are added to some fish sauces during their preparation.6. 123 . the best-known of the fish sauces (gyoshoyu) of Japan is shottsuru. Martin Adams Faculty of Health and Medical Sciences University of Surrey Guildford Surrey GU2 7XH United Kingdom 6. For example. It is produced from the sandfish Arctoscopus japonicus and has the rice-based enzyme preparation. Fermented fish products are largely confined to east and south-east Asia. in economic terms.1 Fish/salt products such as the fish pastes and sauces tend to contain relatively high levels of salt. typically in the range 15 . producing the fish sauces and pastes. 6.25% and are used mainly as a condiment. the term ‘fermented’ describes a spectrum of processes that ranges from the largely autolytic degradation of fish protein. As with most traditional products that are still produced principally on a cottage industry or domestic scale. added during preparation. whereas in those where a carbohydrate source such as rice or sugar is also added. though some are produced elsewhere. there are numerous variants of some common themes and a host of local names used to describe them. The principal fish sauces and pastes of south-east Asia and their local names are listed in Table 6. Those to which only salt is added tend to be mainly autolytic. Sugar can also be added to the Indonesian fish sauce bakasang and the Thai product nam-budu. Essentially. The relative importance of these two activities depends on the formulation of the product. they are exported to European.1 Definitions As applied to fish products. koji. to those processes in which the activity of lactic acid bacteria (LAB) plays an important part. FERMENTED FISH Prof. the most important of all fermented fish products. North American and other markets and are. The fish sauces are produced on the largest scale.1.I. they can be divided into two categories: fish/salt products and fish/salt/carbohydrate products.
Fermented fish products are of very minor importance outside Asia. fish sauces known as liquamen or garum were ubiquitous condiments for the Romans. a variety of products can be produced within a single country. and pissala. In Europe. to products analogous to other lactic fermented foods such as salami. who had adopted them from the Greeks.1. salty taste. 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.II. may be direct descendants.III. and the more widely known Worcestershire sauce a very distant relative. 124 . made from whole fish in France. A selection of Asian products is presented in Table 6. 6. These have long since fallen from use.FISH AND SEAFOOD TABLE 6. where the bacterial production of lactic acid is a major feature. 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. 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.I Fish sauces and pastes of south-east Asia Country Sauce Amber/brown liquid. made from fish livers in Greece. although garos.
burong-ayungi.III Fish/salt/carbohydrate products of south-east and east Asia Country Japan Kampuchea Korea Laos Products I-sushi. pla-som. pla-chao. e. burong-bangus pla-ra.g. crustaceans Variety of fish Variety of brackish or marine fish 1:4 3 . som-pa. mam-pa-kor. cencalok burong-isda.4 months Condiment and main dish Whole 1:3 18 months Condiment Whole 1:3 After 3 .12 months 10% raw sugar added.3 months Main dish Tai-pla Bowels 1:3 6 . salt/fish ratio Production period Type of use Hoi-dong Molluscs Whole or without shells Whole - Main dish Kapi (fish or shrimp paste) Nam-pla (fish sauce) Nam-budu Small fish. funa-suchi. burong-dalag. som-pa-keng pekasam. ayu-sushi. pa-chao. pa-khem. e.8 months Main dish TABLE 6. som-fak/ som-fug 125 Malaysia Philippines Thailand .FERMENTED FISH TABLE 6. shrimps.g. the mixture is then boiled and bottled - Condiment Nam-khoei Shrimps. mam-chao. mam-seeing sikhae som-kay-pa-eun. crustaceans Mackerel eviscerated fish Variety of fish Whole 1:4 Condiment Pla-thu-khem Whole 1:3 2 . tai-suchi phaak.II Fermented fish/salt products common in Thailand Name Type of fish Form Approx.
The diversity of fish used in fermented products (see. rakefisk or surfisk. in Sweden.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. paddy ponds beside rice fields used for collecting fish when the field has dried up. relatively mild-tasting product. A modern account of their production. for example. especially in fish/salt products. some fermented fish products remain popular. made from herring. Norwegian gravlaks. 126 . 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. which has been increasing in popularity over the last 50 years.FISH AND SEAFOOD In Northern Europe. which employ lower levels of salt than Asian products. For example. or buried salmon. it was found that several different sources were used for the fish . malt or flour (1). and the initial microflora is likely to be considerably larger in these cases. freshwater fish are the usual raw material. is a traditional.the flooded rice fields. Here. the most popular varieties of which are rakørret. These are often described as fish/salt products.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. Penaeus indicua Many of the fish used in fermentation are uneviscerated.IV) and in the way in which they are handled prior to processing means that the initial microbial levels are far from uniform. and their microflora tends to reflect their local environment more than that of marine species. states that sugar is often included with the salt. This could lead to quite marked variations since the same fish can be obtained from a variety of sources. Table 6. TABLE 6. however. 6. fermented trout. and counts ranging from below 104 /g to in excess of 107 have been reported. and old descriptions of their production often refer to the addition of whey. and a large local freshwater reservoir (2). in Norway and surstrømming. There are also more heavily fermented products. in a survey of fermented fish products in north-east Thailand.
(3.1 – 6.3. in many cases. Mucor. they can contribute to the initial microflora. Generally. its contribution is likely to be insignificant compared with that of the fish microflora in terms of both spoilage potential and public health concerns. Studies have shown that the number of bacteria declines rapidly during the production of the Thai fish sauce nam pla and that. 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.4. work with nam-pla in Thailand has isolated Halobacterium and Halococcus spp. In another microbiological study of four fish sauces. It also demonstrated their significant proteolytic activity during the first month of fermentation. however. Bacillus spp.FERMENTED FISH Where other ingredients are used.74 or below. 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. where the total counts dropped from above 107 cfu/g to below 103 cfu/g after 14 days and below 102 after 40 days. It has also been shown that a perfectly acceptable patis could be produced from fish that had been sterilised with ionising radiation (6). mean that microorganisms have been generally held to play little or no role in the process that follows. and studies 127 . 6. and the anaerobic conditions that prevail in the bulk of the material. the product contained about 500 cfu/g comprising mainly Micrococcus and Bacillus spp. traditional starter culture preparations such as koji (Japan). Endomycopsis and Saccharomyces . principally species of the genera Rhizopus. In some cases. which included nam-pla and patis. These preparations contain a mixed microflora of moulds and yeasts. the Philippine fish sauce.3 6. 4). in numbers reaching 108 /ml after 3 weeks of fermentation but declining thereafter. were found to be the predominant isolates. angkak (Philippines and Thailand) and look-pang (Thailand) are used. Similarly microbial inactivation by irradiation has not been found to adversely affect the properties of Korean fermented squid (7). More recently. suggesting that halophilic bacteria do play an important role in the production of fish sauce (8). however. sufficient to saturate the water phase present and produce an aw of 0. Similar results were seen with patis. after one month.organisms that play an important part in the production process but can also contribute to the ultimate spoilage of the product. or boiled or roasted rice. This. Hansenula. probably reflecting their ability as spore-formers to survive rather than any capacity to multiply under the prevailing conditions (5).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. Other halophilic species have been identified from fish/salt products. When this is a simple carbohydrate source such as sugar. either directly or as the partially saccharified rice they produce.
6.2. Fig. 6.1. Belacan (Condiment) Fig.12). Patis (Condiment) 128 .FISH AND SEAFOOD have demonstrated the production of halophilic proteinases from fish sauce isolates (9 .
FERMENTED FISH Fig. Myulchijeot (Staple food) The extensive proteolysis and liquefaction that occur during production are.3. and muscle cathepsins have been shown to be inhibited at relatively low salt concentrations (13. In fish sauce production. eviscerated fish are sometimes used to slow the autolytic process. thought to be largely the result of autolytic breakdown of the fish tissues. 14). 6.6.4. however. Ika-Shiokara (side dish) Fig. To produce semi-solid fish pastes as opposed to fish sauces. the 129 . This is more rapid when whole fish are used since the head and viscera contain higher concentrations of proteolytic enzymes than other tissues.
Figure 6.5).6 – 6. filtered and bottled. Fig. snack) 130 . 6.8. particularly glutamate. In contrast to free-swimming fish. which is eventually separated from the residual fish tissues. Their main feature is that the provision of a source of carbohydrate allows a lactic acid bacterial fermentation to occur. 6.2 Fish/salt/carbohydrate products of Asia This category encompasses an even greater diversity of products than the fish sauces and pastes.V and Figures 6.3. Some examples of products and their formulation are presented in Table 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. for example. volatile fatty acids and nucleotides. shellfish such as mussels often have appreciable levels of carbohydrate in their tissues and in these cases addition of carbohydrate is not necessary for a lactic fermentation to occur (see.FISH AND SEAFOOD high salt level used also assists in the osmotic extraction of a supernatant rich in amino acids.5. Hoi-Malangpu-Dong (side dish.
side dish) 131 . north-east. 3:1:1-3 Fish:salt:boiled rice: garlic 10:2:1:0. S.10 days 5 days Main dish Main dish Main dish Condiment and main dish Note: Products 1 .5-1. Fig. Ang-kak: red mould rice obtained by fermenting rice with Monascus purpureus. south of Thailand.FERMENTED FISH TABLE 6.4 are also produced using shrimps. NE 10 . 3:1:0.12 months Shelf life Use 1 .03 Preparation time 6 . C NE.6. N.3 years Condiment and main dish 2-3 months 3 weeks 2 weeks 2 weeks 6 . central. north. NE. Kung-Chom (staple food. N S 5 .2-4 Fish (pieces): salt:khaomark (fermented rice). NE Formulation Fish (whole or pieces): salt: roasted rice. C NE.5:2-3:1 Fish:salt:boiled rice: ang-kak 3:1:3:0. In these cases.12 days 3 . N.V Fish/salt carbohydrate products of Thailand Name 1 Pla-ra Region C.7 days 5 . C. N. the prefix 'pla' is replaced by 'kung'.12 months Main dish 2 Pla-jao C.6.25-1 Fish:salt:roasted rice: garlic 10:1:3:1 Fish (minced):salt:rice: garlic 10:0.20 days 3 Pla-som 4 Pla-jom 5 Som-fak 6 Pla-paengdaeng NE.
but there is considerable variation in this. It also indicates that. then this will take longer to develop than if lower salt levels are used. their growth and metabolism are nonetheless slowed.FISH AND SEAFOOD The extent to which lactic fermentation occurs depends largely on the salt content.VI). (from Arroyo et al.3.1 The use of higher salt levels results in a longer production phase but a better keeping quality product.6. The LAB isolated from these products are those with more marked salt tolerance but.salt and lactic acid. A survey of fish/salt/carbohydrate products in Thailand (15) noted that there were two empirical rules governing formulation:6. 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. 106) Fig.. these products are salted at lower levels than the fish sauces and pastes.10%. Generally.2. 2. giving a reduced rate of acid generation. of the two factors contributing to shelf life . If acidity is a desired sensory property of the product. Red burong isda 132 . while they are able to grow at high salt levels. typically in the range 2 . Phillipines Journal of Food Science and Technology. This is readily explained by the susceptibility of LAB to elevated salt concentrations. the salt is of far greater importance in determining the product’s long-term stability.7.
0 .188.8.131.52 3.3 5.5. White balao-balao TABLE 6.3 years 1 .2 4.9 .9 4.4.8 2. 106) Fig.3 .FERMENTED FISH (from Arroyo et al.5.5.6. 2. Philippines Journal of Food Science and Technology.4 .9 4.4. 133 .6 3.8 pH 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 .0 ..2 years 2 weeks 2 weeks Source: A Concise Handbook of Indigenous Fermented Foods in the ASCA Countries (16).12 months 1 .5 .7 .2 4.5.1 .6.0 Shelf life 3 weeks 6 .5 .1 4.8 .184.108.40.206 .2 7.
also serves as a valuable source of the more readily fermented carbohydrate.1 . efficient lactic acid production will depend upon the presence of amylolytic LAB or saccharification by other microorganisms or enzymes that may be present. a number of amylolytic species have been described and an amylolytic strain of Lactobacillus plantarum has been isolated from burong bangus in the Philippines (17). although the final population of LAB may be similar.VII Fish/salt/carbohydate products in which saccharified rice or a saccharifying starter culture is used Product red burong-isda i-sushi. they increase the amount of soluble sugars produced and thus the range of LAB that can grow. a saccharifying agent such as koji (Japan) or ang-kak (the Philippines) may be added separately (Table 6. The effect of this will be to decrease the product’s final pH and increase the amount of total acidity. Where they are used. a survey of burong-isda in the Philippines reported consistently lower pH values for red burong-isda where ang-kak was used (pH 3.9) compared with the white product (pH 4. inulin. 19). also lower the unit cost of the product and this is probably a significant factor in many cases. of course.FISH AND SEAFOOD 6.3. This results in a more active lactic fermentation and a lower pH. such as salt content. being similar (18. alternatively. For example. the rice may be added in a partially saccharified form such as kao-mark (Thailand).5) despite other factors. In several products.VII).3. Garlic. Although the ability to ferment starch is not widespread in the LAB. a common ingredient in these products. Addition of carbohydrate will.2 Inclusion of more carbohydrate gives a faster fermentation and a stronger acid taste Increasing the proportion of carbohydrate in the formulation decreases the product’s buffering capacity and provides the LAB with more substrate on which to act.2.0 . 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. TABLE 6.4. 21). Rice is the most common carbohydrate source used. When added as boiled or roasted rice alone. 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 .
It seems that a lactic fermentation takes place in many types of pla-ra but is not significant in others. 135 . the pH drops from 6.5 in the first 2 .3 days. the salt levels in the latter are sufficient to rule out microbiological spoilage in all but the most exceptional of cases. 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). and there is a marked regional variation in the preferred type. presumably correlating with the salt content. Results of a product survey were consistent with this. around 12%. reporting the product’s pH to range from 4. Clearly. Cleaned pieces of eviscerated fish flesh are salted with 6% salt overnight before mixing with cooked millet. red pepper powder and garlic in the ratio fish:millet:pepper:garlic of 80:7.4 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).2 at the end of fermentation. Excessive growth and acid production by the LAB could render products too sour for acceptability. Traditional packing techniques used with lactic fermented fish products. which is sufficient to prevent lactic fermentation by all but the most halotolerant LAB.17. The mixture is allowed to ferment for 2 .7 up to 6. Generally. There is little published information on the factors that limit the shelf life of the low-salt products. the level of salt employed is on the high side for fish/salt/carbohydrate products. 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. The Thai product pla-ra exemplifies these basic rules as well as the difficulties associated with attempts to classify such products.9%. the Korean fermented fish product.7 to below 4. will not reliably exclude air. and surface mould growth has been shown to be a cause of rejection in the Thai product som-fak (24). The growth of yeasts and moulds could. allowing other spoilage organisms to grow. lead to an increase in pH. thereafter declining to about 4. Any loss of acceptability must arise from chemical or physical changes occurring over prolonged storage.2. One study found that any effective measure to exclude air from a fermenting fish/carbohydrate mix prevented yeast growth (23). but they are likely to behave similarly to lactic fermented products produced from other commodities. such as wrapping in banana leaves.3 weeks at around 20 °C. but in most cases it is probable that yeast and mould growth would be an important cause of spoilage. This suggests that use of oxygen-impermeable wrapping films could extend the shelf life of such products.FERMENTED FISH Sikhae. is something of an exception since millet is used along with garlic as the carbohydrate source.5:9:3. In a successful fermentation.5. in turn. 6. A combination of irradiation and chill storage has been shown to prevent both over-acidification and fungal growth in som-fak. markedly extending the shelf life (25).8 . which ranged from 7.
The fish/salt/carbohydrate products depend on the combination of salt and pH/lactic acid for their safety. outbreaks of botulism are the only well documented instances of illness associated with fermented fish products. Their safety therefore is critically dependent on the quality of the raw materials used and the inhibitory effects of the fermentation process. As in Norway. The combinations of pH and salt in many fermented products are likely to preclude growth of the organism completely and.5 Pathogens: Growth and Survival Many fermented fish products are stored at ambient temperatures. fish products are the most common source of botulism. During fermentation.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. but we are unaware of any reports of this happening. which accounted for 88% of outbreaks in the period 1952 . and the most frequently implicated product is the fish-salt-carbohydrate fermented product Izushi. After capture. are Vibrio parahaemolyticus and Clostridium botulinum. the pH decreases below 5 and safe processing is recognised to depend on proper salt levels and temperature control until the pH drops. 6. outbreaks of 136 . In Japan. under conditions inimical to growth and high ambient storage temperatures. V. however. and failure to achieve a sufficient combination of inhibitory factors could lead to their growth or survival.8 and at salt levels as high 10%. parahaemolyticus has been reported growing down to a pH of 4. fish can potentially be contaminated with almost any foodborne pathogen as a result of improper handling and storage. The Scandinavian fermented fish product rakefisk was responsible for 16 (84%) of the outbreaks of botulism recorded in Norway in the period 1961 90. although these limits apply when all other factors are optimal (26). Because of the severity of the illness it causes. and consumed without cooking. The bacterial pathogens most commonly associated with the living environment of fish. The epidemiological evidence points to greater awareness of the importance of these factors since outbreaks have declined over recent years (27).87. typically 30 °C. 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. There is little in the way of documented information associating fermented fish products with foodborne illness. Several measures have been suggested for reducing the risk 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. viable numbers will probably decline quite rapidly. Improper storage of fish prior to processing could result in the production of toxins capable of persisting through to the product. In the fish sauces and pastes.
105-10.. 31.. Liston J. Thailand. 2. Infection with the trematode Opisthorchis viverrini is associated with cholangiocarcinoma. they could act as an important vehicle of transmission. and salmon heads and eggs. Rakefisk and Herring in the Norwegian Tradition. Institute of Nutrition. viverrini were a strong risk indicator and that consumption of fermented fish products was an independent risk factor in the contraction of cholangiocarcinoma (31). Novus Press. botulinum and the production of toxin. Microbiology and chemistry of fermented fish. 137 . Freshwater fish are the most important intermediate reservoir hosts of foodborne trematodes. beaver tail and paw. They are packed into containers such as buckets or pits in the ground and left at ambient temperature to putrefy.. Oslo. An investigation of survival of the fish parasitic nematode Anisakis in marinated herring has given some indication of what these studies might find. 1989. Yhoung-Aree J.O. One study with the trematode Haplorchis taichui. Kasemsarn B. and warm weather can result in rapid growth of C. Salting levels are often insufficiently low to prevent growth of C. Botulism has also been associated with a number of traditional foods of the peoples of northern Canada.M. though 27% survival was still recorded (33). walrus nose. Raw materials subject to this process include seal flipper... Saisithi P. With this particular organism. 6. The more widespread current use of non-traditional containers such as glass or plastic jars is thought to be one contributing factor (28). although the absence of any source of fermentable carbohydrate means that the process is putrefactive. A case-control study conducted in northeast Thailand showed that elevated levels of antibodies against O. whale meat. Foodborne trematode infections are emerging as a major public health problem. Where these products are consumed without final cooking. particularly in south-east Asia and the western Pacific regions (30). Journal of Food Science. Greenland and Alaska. and these are often used in the production of fermented fish products. affecting an estimated 40 million people. 1966.FERMENTED FISH botulism have become less frequent in recent years. Many of these products are described as fermented. Dhanamitta S. probably reflecting greater awareness of the critical factors that determine the product’s safety (27). Mahidol University. Fermented Fish Products Study (Thailand) Phase 1. which will persist into the product. salt content was found to have the greatest impact on the viability of larvae. giving an alkaline fermentation. Dollar A. showed that inclusion of fermented fish in the Thai dish lab-pla improved inactivation of the metacercariae. Puwastien P. while increasing the acetic acid content of the formulation had little or no effect (32). which is generally consumed raw (29). botulinum. References Riddervold A. Studies to determine the effect of traditional processing techniques on the infectivity of metacercariae was identified as a priority research area by a WHO study group (30). 1990. Baseline Study (1987-1988). 3. Lutefisk.6 1.
Im W. Jae-Hun K. Olympia M. Crisan E. Hiraga K. Bioscience. Yasir M.. Process Biochemistry... Tanasupawat S.M.. Nov. Catheptic activity of fish muscle. 80. Dong-Ho K. Tropical Science. Nov. Rattagool P. Phil. 29. microbiological and chemical properties of salted and fermented squid.. 138 . Kudo T. A Concise Handbook of Indigenous Fermented Foods in the ASCA Countries. Tanasupawat S. 2007.. Saono S. 106-8. 1986.. 6338. Thongthai C.H. Suntinanalert P..S.H. Fermented fish products of south east Asia. Lentibacillus kapialis spp. Plenum Press.. Applied Microbiology. 57... 1395-401. 1995. Constantinides S. 12. A halophilic serine proteinase from Halobacillus spp. Production and characterisation of NaCl-activated proteinases from Virgibacillus spp.J. Yook H.M.. Journal of Food Protection. Lim J.. 934-9. 1972. Lee S. Sands A. isolated from jeotgal.. New York. Canberra. Thawai C. 1975. Protein hydrolysate from fish. Dhamcharee B.. Dymsza H. Some chemical changes in fish silage. Pakdeeto A. 43.. Sinsuwan S. 1985. 141-2. SK33 isolated from fish sauce fermentation. Aslam Z. 9. Namwong S. Yongsawatdigul J. Liston J. International Journal of Systematic and Evolutionary Microbiology. Agents of proteolysis and its inhibition in patis (fish sauce). Cooke R.FISH AND SEAFOOD 4. Orejana F. Government of Australia. Takano M. 10. 11. 2008.A. Microflora of four fermented fish sauces. 61-73. 15.. 5. Journal of Food Technology. 3649. Halophiles in Thai fish sauce (Nam pla). 57. Rodriguez-Valera F..K. 37. 17-30. Moonmangmee S. from fermented shrimp paste in Thailand.. 353-63.. Ono H. 25. Ed.R.. Itoh T. Reddi P.. 2000.. 16.V.T. Journal of Scientific and Industrial Research (India)...D. 643-7. 63. in General and Applied Aspects of Halophilic Microorganisms. Rodtong S. Effects of gamma radiation on sensory qualities... Characterisation of starch-hydrolysing bacteria isolated from a fermented fish and rice food “burong isda” and its amylolytic enzyme...T. Salinococcus jeotgali spp.. 7. International Journal of Systematic and Evolutionary Microbiology. SR5-3 isolated from fish sauce: purification and characterisation. Kyong-Haeng L.. 11. 1957. Chung Y. 17... 8. 381-8.K. Adams M. Biotechnology and Biochemistry. 185-92. Journal of Food Science. Tsunemi M. Backhoff P. Ahn H. a traditional Korean fermented seafood. 16A.. 3.W. Oda K. 6.. 2007. Byun M. Takada K. 1976. 14. 70.R.R. 1991. 2006.. 13.. Journal of Fermentation and Bioengineering. 1979. Velankar N. Journal of Food Science and Technology. Fukuda H. Hull R. Kaneko Y. 124-30.
1998. 25. Gram L. The influence of oxygen accessibility on the growth of yeast in fish/rice fermentation.FERMENTED FISH 18.. Saisithi P. Tanaka M.. 1815. Huss H. 103. Riebroy S. Owens J.D. Food Chemistry.A... Blackie.W.. Martin J. Moody L. Characterisation of lactic acid bacteria isolated from a Thai low-salt fermented fish product and the role of garlic as substrate for fermentation. 28. 22... 3-10. Control of foodborne trematode infections. Ed. Wedell-Neergaard C. Patrick D... Canada Communicable Disease Report..H. 3. Journal of Agricultural Science (Tokyo). Fung C. Paludan-Müller C... Caldo G. Sakai H. Paludan-Müller C. 2002. burong-isda. 28.. 105-12. WHO Study Group. Marcel Dekker. Hauschild A.. Ben Embarek P. 307-14. 305-22. Hazards from northern native foods. 1983.D. 46. Traditional fermented fish products with special reference to Thai products. in the Philippines. 30.. 1996.. 2002. a Thai low-salt fish product. 1-4. in Clostridium botulinum: Ecology and Control in Foods. 2007. 28 (6). 20.. Isaac-Renton J. Hauschild A. 139 . Kozaki M. Marcel Dekker. Hauschild A. World Health Organisation Technical Report Series. Caldo G.L. The fermented fish food. New York. 426-35. Huss H. Visessanguan W.. Journal of Food Science and Technology. 219-29. 26. 92. Huss H. 1992.. Gram L. 69-104. Journal of Applied Microbiology. Dawar M. Epidemiology of human foodborne botulism. a Thai low-salt fermented fish product.H. International Journal of Food Microbiology. ASEAN Food Journal. Fermentation and spoilage of som fak. 1992. Dodds K. Genotypic and phenotypic characterisation of garlic-fermenting lactic acid bacteria isolated from som-fak. International Commission on Microbiological Specifications for Foods.W.. 21. 1983. 138-44.K. New York. in Clostridium botulinum: Ecology and Control in Foods. Effect of irradiation on properties and storage stability of Som-fug produced from bigeye snapper. 28.L. 23.H. Avurhi J. Valyasevi R. 27... 274-86..W. Ostergaard A. Kozaki M. 29. Vibrio parahaemolyticus. 27.. 104-6.H.H. Eds. 19.B. London. in Microorganisms in Foods. Dodds K. Erikson U. Eds. Gram L. Volume 5: Microbiological Specifications of Food Pathogens. Two outbreaks of botulism associated with fermented salmon roe – British Columbia. Rustad T... Journal of Agricultural Science (Tokyo). 1987. Yamprayoon J. Tropical Science. 24. Benjakul S. 1995. International Commission on Microbiological Specifications for Foods.H. Wainwright R. 1990.. Yeast-flora in red burong-isda a fermented fish food from the Philippines.M. 1999. 38.A. Sakai H.B.
Southeast Asian Journal of Tropical Medicine and Public Health. Honjo S. Ruamsuk A. Horst K. Bloemsma B.H. 15998. 23.. 36. 1995. Aspen Publishers Inc. 117. Fermentation and Food Safety.7 Further Reading Adams M. 2001..J. Jedpiyawongse A. Huss H. Sriplung H..R. 854-60. Uchida K... Survival of Anisakis larvae in marinated herring fillets.. Deerasamee S.. Wongsawad C. Nithikathkul C. Kikukawa H. Srivatanakul P.R..R. 32. 110-1. Adams M. 140 . Gaithersburg. Hanai S.. Fermented flesh foods. 661-70. Roepstorff A. 1986. Survival of Haplorchis taichui metacercariae in lab-pla.... 33. International Journal of Food Science & Technology..... Progress in Industrial Microbiology. Chuboon S. Sripa B. Miwa M. Nout M. Genetic and environmental determinants of risk for cholangiosarcoma via Opisthorchis viverrini in a densely infested area in Nakhon Phanom. Kittiwatanachot P. 2005. 6..FISH AND SEAFOOD 31. 29. northeast Thailand. Todoroki T. International Journal of Cancer. Thai traditional food preparation. 2005..
including the well known marine toxins 141 .1 Introduction Shellfish and finfish toxins continue to be a threat to public health and local fisheries around the world. produced by the Intergovernmental Oceanographic Commission (IOC) of UNESCO (United Nations Educational.g.D. Scientific and Cultural Organisation) has been updated (7).C.. Nevertheless. The manual on harmful marine algae and their products. Liquid Chromatography/ Mass Spectrometry/ Mass Spectrometry -LC/MS/MS) is employed to identify and confirm the presence of known toxins. Since the original chapter in this series was published (6). several scientific reports have described their efficacy and potential. and a second updated edition dealing with the pharmacology. and is often necessary in light of the complex toxin profiles that can occur in shellfish tissue. known and unknown. the export of exotic seafood that may contain such toxins can lead to unexpected public health issues. FCIC UNC Wilmington Center for Marine Science Marvin Moss Lane Wilmington NC 28409 United States of America 7. Wright.3) and some practical applications for detecting shellfish toxins have been reported (4. physiology and detection of marine and freshwater toxins has also appeared (8). 5). Ph. a number of new toxins and toxin families have been identified. In the last decade seafood monitoring has intensified and detection methods have continued to develop in selectivity and sensitivity. Although most of these antibody-based test kits have yet to receive international regulatory approval. C. While some of these toxins only occur in seafood from a restricted geographical region. more traditional methods such as the mouse bioassay continue to be used in many countries to warn of the presence of toxins. Another important development is the introduction of sensitive and selective antibody-based detection methods for a number of toxins. It remains to be seen if this approach gains momentum in the future.. Another book dealing with the clinical effects of a variety of poisons and toxins. An emerging technology is the use of immunosensors (1 .7. FISH AND SHELLFISH TOXINS Jeffrey L. most notably the azaspiracids and the cyclic imines as well as some additional polyethers.M. Increasingly sophisticated instrumentation (e.
2.1). In addition. 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. The remarkable frequency of neurotoxins from marine dinoflagellates has been noted (13). and sub-tropical waters and habitats (15). Around twenty PSP toxins are known. Not all the toxins found in shellfish extracts are microalgal products. with intoxications occurring in many countries even today (15). a great variety of shellfish from all latitudes.2 Paralytic Shellfish Poisoning (PSP) Toxins The PSP toxins are arguably the most recognised group of marine toxins and continue to be regarded among the most dangerous. can become contaminated with these toxins. neoSTX and Gonyautoxins (GTX) 1-4.1 Origins and distribution The PSP toxins are produced by several dinoflagellate species belonging to the genera Alexandrium. Gymnodinium catenatum. The Ncarbamoyl sulfates are readily hydrolysed to STX.FISH AND SEAFOOD has been produced (9). dealing with the detection methods and regulatory requirements for many marine toxins has been published (12). All PSPs degrade rapidly in alkaline conditions. Many of these species are extremely common and are found throughout the world in a variety of cold. Consequently. Saxitoxin (STX) and neosaxitoxin (neoSTX) are the parent members of the group and are the most toxic. and a review on the risk assessment of marine toxins has also been produced (14). and a tropical species Pyrodinium bahamense (16). 7. An account of this meeting detailing recommendations and scientific advice on marine biotoxins has been published (10). 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. some are the result of biotransformation processes by the filter-feeding host. temperate. all of which contain a tetrahydropurine skeleton (Figure 7. A brief report on the status of rapid detection methods for phycotoxins in the European Union (EU) has appeared (11). 7. even crustaceans such as lobsters. Saxitoxin Group instead of the Paralytic Shellfish Poisons). PSPs are watersoluble and heat stable so cooking has little or no effect on PSP toxicity.g. 142 . and another.
and fingers within 5 .2.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.2 Toxicology PSP toxins affect the central nervous system of mammals. this is never sufficient to render the seafood safe to consume. leading to paralysis within a few hours and in extreme cases death by asphyxiation due to respiratory paralysis. Neurophysiological studies have shown that PSP toxins all possess the same mode of action though some derivatives such as STX and neoSTX are considerably more potent than the C-toxins (15). PSPs bind to site 1 of sodium channels reversibly blocking sodium conductance in nerve and muscle membranes by binding to a specific receptor site located on the outside surface of the membrane close to the ion channel opening. However. Cooking does not destroy these compounds and although some of the toxins may be extracted into the cooking water. 7.30 minutes of consumption. Structure of saxitoxin and neosaxitoxin and their sulphated derivatives 7. and humans seem to be particularly susceptible. tongue. 143 .1. the less potent C toxins are readily hydrolysed by acid to the more potent GTX toxins. Human symptoms begin with tingling and numbness of the lips.
as well as other parts. and while simple and relatively inexpensive to operate. China. a pre-column method has received Association of Analytical Communities (AOAC) approval (18). or conversion before HPLC separation (pre-column derivatisation) and these have recently been thoroughly reviewed (12. a criticism has been that such methods are time consuming. and has also been evaluated by a number of EU laboratories. namely oxidative conversion to fluorescent derivatives after High Performance Liquid Chromatography (HPLC) analysis (post-column derivatisation).FISH AND SEAFOOD 7.3 Tetrodotoxin Tetrodotoxin (TTX) possesses a unique chemical structure and was the first marine toxin to be chemically characterised. After some development and various trials. Two approaches have been taken here.2. The mouse bioassay has been in place for decades and the method has been standardised. including Japan. 17). other derivatives have been reported. Alternative HPLC methods have subsequently reduced this to a single HPLC run. TTX most commonly occurs in many species of puffer fish. 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. However it remains as the monitoring method of choice in many countries. Thailand.3. specifically 144 . 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. While the chemical detection methods are most promising for the future. There continues to be a significant effort applied to the development of alternative chemical and biochemical detection methods for these toxins. 7.8 µg/g) of shellfish or seafood tissue. and islands of the South Pacific. Since its discovery. One approach has been to take advantage of the ready conversion of these toxins to fluorescent derivatives. 24) (Figure 7. 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. 7.2). and more standards of individual PSP toxins are required to validate the complete profile to toxins that could occur in contaminated shellfish tissue. This kit was found effective in all the samples tested in Europe (20) and was applied recently in a survey of Scottish shellfish (21). and a second rapid dipstick kit (MIST Alert™) is available for the qualitative detection of positive and negative samples (19).1 Origins and distribution Unlike many other marine toxins there is no temporal distribution of TTX episodes. An antibody-based kit that can detect STX in shellfish is commercially available (RIDASCREEN®).3 Detection methods The safe limit set by most countries for PSP toxins is 800 µg/kg (0. Most intoxication events have occurred in South East Asia. Taiwan. 15. 23.
Chemical methods for TTX detection have also been investigated and are mainly based on its smooth conversion by heating in an alkaline solution to a detectable fluorescent (C9) base.2 Toxicology Like the PSP toxins.3. nausea and vomiting. raising speculation that TTX is produced by a bacterial source. and once again the mouse bioassay has been used to detect the presence of the toxin. this has expanded to include bacteria within the Alteromonas and Pseudomonas genera. 25. respiratory failure and coma.3 Detection methods TTX can be extracted from suspect tissue using dilute acid conditions.3. 23. and more recently by an actinomycete Norcardiopsis dassonvillei (26). gonads and roe (24). where puffer fish of the genus Fugu are a delicacy. preventing depolarisation and propagation of action potentials in nerve cells. and ingestion of larger amounts leads to cardiovascular effects including bradycardia. 7. Structure of tetrodotoxin 7. the blue-ringed octopus. horseshoe crabs. molluscs. and even newts. However TTX has been found to occur in a remarkable variety of genetically unrelated species including gastropods. hypotension. TTX acts on the central nervous system by binding extracellularly to site 1 of voltage-gated sodium channels. This was indeed confirmed and a number of marine Vibrios have been shown to be capable of TTX production (24. An oral dose of 1 .2 mg of purified toxin can be lethal. Severe poisoning causes paralysis and respiratory failure. 7. 26). In Japan. A review of TTX detection methods has been published (12. and in mild cases results in distal muscle weakness. and blocks diffusion of sodium through the sodium channel. 27). it is served in speciality restaurants with qualified chefs to properly prepare the fish.2. The onset of symptoms usually occurs rapidly.FISH AND SHELLFISH TOXINS in the liver. Since then. Fig. All of the observed toxicity is secondary to the action potential blockade. Typically this has been accomplished using HPLC followed by post 145 .
but 146 .FISH AND SEAFOOD column oxidation to the fluorescent derivatives. 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. but it is known that such congeners are less toxic than DA itself (30).3 for examples). Surveys by a number of groups have identified over a dozen species of Pseudonitzschia capable of producing DA. and while promising. Fig. Immunoassay methods have also been investigated. but all of them are considerably less toxic than DA itself (29). These isomers arise as a result of isomerisation and location of the double bonds in the side chain of the molecule.3.4 Amnesic Shellfish Poisoning (ASP) Toxins The parent compound of this group is domoic acid (DA). However this method has been criticised for being complex. Structure of domoic acid and three naturally occurring photoisomers 7. Since the original episode of human poisoning following consumption of DA-contaminated shellfish in Canada in 1987 (28). 7. 7.4. and more rapid LC-MS methods [(M+H)+ m/z 320] have been reported for the detection and identification of TTX and its various derivatives (27).1 Origins and distribution The cosmopolitan diatom Pseudonitzschia multiseries was identified as the culprit organism in the 1987 poisoning event (31). need further development (24). a polar glutamate agonist.
33). which has been well documented in studies along the west coast of the US. Victims display a variety of symptoms including headache. Penetration of the blood brain barrier is poor in healthy subjects. Four deaths were attributed to the Canadian incident and years later a few seriously affected individuals still display chronic neurological deficits. 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. cormorants and pelicans (34). and this has led to ASP toxicity in birds. scallops. and clams. as does impaired renal function which increases serum concentration and residence time of the toxin. multiseries. including persistent short term memory deficits (36 . 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. but any impairment of the blood brain barrier results in additional risk. 147 . herbivorous fish become toxic after ingesting strains of Pseudonitzschia spp. The oral absorption of DA is around 5 . Indeed. found in the Pacific around Vietnam. the Philippines. disorientation and coma.FISH AND SHELLFISH TOXINS only three strains (P. and sea animals such as sea lions and even whales (35). nausea. To a great extent. 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.2 Toxicology DA is an acidic amino acid and potent neurotoxin unaffected by cooking or steaming. and is mainly distributed in the blood. the Mediterranean. and in locations around New Zealand (32. another diatom species Nitzschia navisvaringica.4. Levels of DA in the other strains are generally <1 pg/cell. seizures. southern and western portions of Ireland.. For example. 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. 7.10% of the administered dose. Another factor is the movement of the toxin through the food chain.39). Okinawa and more northerly parts of Japan. The effects of DA may be augmented by the excitotoxic effects of glutamate and aspartate in the sample. 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. Recently. has been reported as a medium-level producer of DA (33). Pseudonitzschia australis. and Pseudonitzschia seriata) are considered to be high producers of DA (>10 pg/cell). and review of the toxicologic pathology of DA has been published (36).
and consequently were often overlooked as the real source of the toxins. as can LC/MS. are the more potent source of toxins – often very low cells/ml of seawater can result in human poisoning. Of additional interest is the use of molecular probes originally developed to detect the presence of cells of potentially toxic Pseudonitzschia spp. though this method is most commonly used to analyse plankton and seawater (42). Dinophysis spp. 48. Alternatively. 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.5 Diarrhetic Shellfish Poisoning (DSP) Toxins The active agents responsible for DSP incidents are a group of polyethers that include okadaic acid (OA). Detection levels can be further lowered by conversion of DA to the fluorescent fluorenylmethoxycarbonyl (FMOC) derivative. Since then. Provided the concentrations in shellfish are high enough (>50 µg/g shellfish tissue). DTX-2 has been found to be extremely widespread throughout many European maritime countries and is often the major DSP toxin present. and dinophysistoxin-2 (DTX-2) (Figure 7. DSP toxins are produced by dinoflagellates of the genera Prorocentrum and Dinophysis (47.. The probes have been employed in trial studies in New Zealand (44) and Scotland (43) to detect P. produce a variety of other 148 . australis. the heterotrophic dinoflagellates Dinophysis spp. An EnzymeLinked ImmunoSorbent Assay (ELISA) kit (Biosense) for DA has been approved by the AOAC. In the context of this chapter. The benthic Prorocentrum spp.30 ng/g) with a rapid clean-up step using a strong anion exchange resin. and this is typically the method of choice.FISH AND SEAFOOD 7. To further complicate matters.4). Pseudonitzschia fraudulenta and Pseudonitzschia delicatissima were found to be less effective. Since their first characterisation it was believed that the only causative agents of this group were OA and DTX-1 (45). 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). and these methods have been reviewed (40. but just over 15 years ago a third toxin. are amenable to laboratory culture and consequently are used in many experiments involving studies of DSP toxins. the term “DSP toxins” will only be used to refer to these three polyether compounds and any derivative based on them.4. and an immunosensor method has also been reported (4). The sensitivity of the LC/UV (ultra violet) method (detection limit ~ 1 µg/g) can be improved (20 . DTX-2 was discovered in contaminated Irish shellfish (46). although in the latter study the probes for Pseudonitzschia pungens. 49). 41).. shellfish tissue can be efficiently extracted with aqueous methanol (1:1). 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. 7.3 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. California. in Monterey Bay. dinophysistoxin-1 (DTX-1). However.
In comparison. Dinophysis caudata.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. Lately these compounds. and include Dinophysis acuminata. Structure of okadaic acid and the two related DSP toxins 7. DSP toxins produced by benthic Prorocentrum spp. While these esters are not toxic in vitro. Fig. The parent acids are the only DSP congeners to be found in shellfish tissue. Dinophysis fortii. although occasionally fatty acyl esters linked through the 7-OH group (referred to as DTX-3 compounds) are also found and are likely a result of bioconversion in shellfish tissue.5. are also recognised as a threat to human health. analysis of Prorocentrum cultures have uncovered an additional series of diol esters of OA and sulphated diesters (called DTX-4. and anecdotal evidence now suggests that several seafood poisoning episodes during the 1960s and 1970s in the Netherlands. Today over ten Dinophysis spp. This was probably not the first DSP episode. and DTX-6) all of which are rapidly converted to OA when the plankton cells are ruptured (48). 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.4. DTX-5.FISH AND SHELLFISH TOXINS lipophilic compounds that are toxic in the mouse bioassay.. Dinophysis acuta. such as Prorocentrum lima. and Dinophysis norvegica among others (47. 149 . but do not have the same mechanism of action as DSP toxins nor induce the same symptoms. they can be converted to the parent toxins by nonspecific esterases or lipases either in the producing organism or in the host shellfish. Prorocentrum maculosum. which include the pectenotoxins and the yessotoxins. Norway and Japan were likely due to DSP toxins produced by a Dinophysis spp. 7. 49). and Prorocentrum belizeanum. In this case Dinophysis was suspected as the culprit organism. are recognised as DSP toxin producers.
3 days. Due to the lipophilic nature of the toxins. 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. which can lead to an underestimation of the total DSP content in a sample. Since the late 1970s. 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. Immunoassays have been developed for OA detection but an inherent problem of these methods has been the low cross reactivity with DTX-1. DSP episodes have been reported worldwide and are included in many monitoring programs (47). 7. This regulatory level includes the possibility of LSPs in the DSP fraction being tested. 7. All the DSP toxins have around a thousandfold greater affinity for PP2A than PP1. Human symptoms include diarrhoea. 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). 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). even with the minimal of clean up steps (48). New Zealand and Chile. and a detection limit of 100 µg/kg has been reported (12). In recent years.2 Toxicology The DSP toxins are potent inhibitors of the serine/threonine phosphatases PP1 and PP2A. and abdominal pain. The onset of symptoms can occur within 30 minutes of consuming contaminated shellfish and generally last about 2 . At the first Meeting of the EU National Reference Laboratories on Marine Biotoxins and Analytical Methods and Toxicity Criteria in 1996. A regulatory level of 80 .FISH AND SEAFOOD Perhaps as a consequence of these multiple sources of DSP toxins. DSP episodes have been reported from Japan. nausea. and while this provides critical seafood safety information. vomiting.3 Detection methods Once again the mouse or rat bioassay is relied upon to detect DSP toxins in contaminated seafood. but the method is complex and matrix effects as well as interferences with other lipophilic compounds such as the pectenotoxins compromise the method. most progress in DSP toxin detection has been using new or improved LC/MS methods and a detection limit of 10 µg/kg has been reported. DSP-Check is a commercially 150 . some careful clean-up steps are required to avoid such matrix interferences and to simplify data interpretation (48). it does not provide information on the DSP toxin profile.5.160 µg OA equivalents/kg of whole shellfish tissue has been implemented in some countries. various European countries. 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. Preparation of a fluorescent 9-anthryldiazomethane (ADAM) derivative followed by HPLC offers reasonable sensitivity.
6.FISH AND SHELLFISH TOXINS available ELISA test kit that can detect both OA and DTX-1 with comparable sensitivity. and the latter group are usually found to be responsible for any reported shellfish toxicity (59). they are toxic in the mouse bioassay and can result in false positives and hence are included here for completeness. 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.5 and 7. If a European collaborative study of the fluorometric protein phosphatase inhibition method is successful it may eventually be approved for regulatory purposes in the EU. 55).5. Structure of the principal pectenotoxins and one of the seco-acid derivatives 151 . with a claimed detection limit of 20 µg/kg. As a result these compounds are now being referred to as lipophilic shellfish toxins (LSTs) (49). with good agreement with the mouse bioassay and LC–based methods (54.6 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). However. In yet another approach. 7. fluorometric protein phosphatase inhibition assays have been found to perform better than colorimetric assays. 58). Some representative examples of both groups of toxins are shown in Figures 7. but with the increased availability of pure toxins permitting detailed toxicology studies it is now recognised that both these groups of compounds possess a different toxic profile from the DSP toxins. Indeed. 7. 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. and another rapid antibodybased test kit for DSP toxins has also been reported (53). Within this group.
2 Toxicology PTXs show varied toxicity in the mouse bioassay (59). While 152 .6. many parts of Europe and the Mediterranean.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). 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. 57. 58). PTX-1 and PTX-2 are frequently the most common derivatives to occur and PTX-2 is the most potent following intraperitoneal (IP) injection (230 µg/kg body weight). which may co-occur in blooms with other organisms. the toxin content and profile may vary among different strains. Structure of the two parent yessotoxins. New Zealand and South America (49. but nevertheless the toxins are extremely widespread and have been found in various shellfish in Norway and Scandinavia. 7. The sources of YTXs have now been identified as the dinoflagellates Protoceratium reticulatum and Lingulodinium polyedrum. Japan.6.6. YTX and homo-YTX 7. eastern Canada.FISH AND SEAFOOD Fig. 7. As is typical for many algal species.
Although there is no evidence that yessotoxins are responsible for human illness. There are two skeletal types known as PbTx-1 (or brevetoxin A) and PbTx-2 (brevetoxin B) (Figure 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.7 Neurotoxin Shellfish Poisoning (NSP) Toxins NSP episodes are caused by a family of fused polyether compounds known as the brevetoxins (63. While LC/UV detection is possible for PTXs. but less so following oral dosing (61).FISH AND SHELLFISH TOXINS demonstrating considerable toxicity following IP injection. The crude extract may be used directly in the mouse bioassay. The structures and chemical data including the molecular weight of each member have been collated (56). but a fluorescent derivative has been prepared for others. Some of the PTXs are potent cytotoxins in vitro (60).6. 7. The YTXs are retained in the aqueous methanol fraction while the PTX and DSP toxins are concentrated in a dichloromethane fraction. which is further compounded by differences in the experimental design. 58). Estimates of acute toxicity of the parent compound following IP injection vary considerably. some of which are found as glycosides and sulfates. Acetone extracts of shellfish tissue are dried and re-suspended in 60% aqueous methanol before partitioning against dichloromethane. There are no clear cases of human illness resulting from ingestion of PTXs (59).7). chemical analysis is best carried out by LC/MS and several approaches have been described (57. the PTXs show considerably less toxicity when administered orally. and this is believed to be due to hydrolysis of PTX-2 to the seco acid derivative. For many years it was believed that 153 . though its cell effects may be linked to modulation of calcium channels (62). 64). it may be compromised by other biological materials in the matrix. 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. a regulatory level of 1 mg/kg shellfish tissue has been set. However. The YTX group has not been reported to cause human illness. the PTXs are susceptible to bioconversion in shellfish tissue and consequently it is often desirable to determine the profile of toxins present. Because of the number and complexity of the YTX group. which aids detection by LC/UV. but various congeners are acutely toxic in the mouse bioassay following IP injection. Their toxicology and mechanism of action remain uncertain. Many YTX toxins contain a conjugated diene system. 7.3 Detection methods Typically these toxins are co-extracted with DSP toxins following acetone or aqueous methanol extraction of the shellfish tissue.
66). and Heretosigma akashiwo.7. PbTx-1 and PbTx-2. but in 1992/1993 they were identified as the culprits in a human poisoning event in New Zealand (65. A complicating factor both in terms of establishing the profile and toxicity of brevetoxin-contaminated shellfish has been the discovery of the so-called “brevetoxin metabolites”.7. the same toxins have been reported to be produced by three genera of Raphidophytes. In the Gulf of Mexico. They have also been associated with fish kills along the eastern mid-Atlantic coast of the US. brevetoxins are found in mussels. whelks. Interestingly. 66). and one of the brevetoxin metabolites BTX-B2 7.S. was chemically characterised as PbTx-2 (67). which are found in Japanese waters (66).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 cockles (65. Fibrocapsa japonica. These are biotransformation products of principally PbTx-2 by addition of certain amino acids to the conjugated aldehyde function (66). 154 . and off the coast of New Zealand (65). oysters. Structure of the two parent brevetoxins. verruculosa off the mid Atlantic coast of the U. Fig.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. namely Chatonella marina. 7. Many of these species are associated with massive fish kills.
In this application. but this method is now used routinely in New Zealand and the US to monitor for the presence of brevetoxins (68). shellfish homogenates. malaise. the competitive ELISA method correlated most closely with the mouse bioassay results.FISH AND SHELLFISH TOXINS although recently it has been shown that herbivorous fish can store brevetoxins in their flesh (68). and it has also been used in studies to quantify occupational and recreational exposure to aerosolised brevetoxin exposure (73). The mouse bioassay is used as the primary detection method. this family of toxins has proven to be a challenge both to identify and monitor.7. In a comprehensive multi laboratory evaluation of five different detection methods as possible replacements for the mouse bioassay. A second generation competitive ELISA assay has been developed that has successfully quantified brevetoxins in a variety of biological and environmental matrices including seawater. While the mouse bioassay continues to be used. nausea. The human health implications following exposure to NSPs have been reviewed (69). 7. The brevetoxin metabolites are reported to be less toxic than the parent compound (70). with a mean duration of symptoms of around 1 day. Ingestion of brevetoxins results in a variety of symptoms including parathesia. 69).2 Toxicology Brevetoxins bind to site 5 of voltage-gated sodium channels. Ciguatera toxins (CTXs) are extremely potent and occur in finfish rather than shellfish.8 Ciguatoxins Known for centuries and originally referred to as ciguatera by Spanish explorers in the Indo-Pacific ocean and Caribbean.7. 7. and mammalian body fluids without pretreatment or dilution (72). 7. vertigo. The mean time to onset of symptoms is around 3 hours.3 Detection methods Three basic approaches for the detection of NSP toxins have been recently reviewed (71). 66. beginning with herbivores that feed on benthic dinoflagellates associated with reefs and which are 155 . with hot spots in the coral reef areas of the Caribbean and French Polynesia. pre-treatment or clean-up of the shellfish sample is essential in order to obtain reproducible results. Exposure to brevetoxins can also occur through inhalation of brevetoxin aerosols resulting in irritation of the upper respiratory tract (65. and diarrhoea. and in severe cases seizure. considerable progress has been made in the development of LC-MS methods to routinely monitor for brevetoxins in shellfish. 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. Outbreaks of ciguatera poisoning are confined to tropical and sub-tropical regions of the ocean. which can result in a cascade of associated events in a cell (66).
7.FISH AND SEAFOOD transferred through the food chain to the larger carnivores. A detailed comprehensive review of ciguatoxins has recently been published (74). complex. Structure of the Pacific variant of ciguatera toxin (P-CTX-1) and the Caribbean variant (C-CTX-1) 7. most likely a consequence of biotransformation of the parent toxins as they pass up the food chain. green and brown seaweeds and also occur in sediments and coral debris. but more are suspected to exist. and other genera of neighbouring reef dinoflagellates can further contribute to an extremely complex profile of toxins in finfish. Indeed even the chemistry of CTXs from the Pacific differs subtly from those found in the Caribbean (Figure 7. CTXs are large. The diversity of benthic phytoplankton that exist on reefs further complicates the picture. a group of closely related polyethers that are considered the source of ciguatoxins.8. Distribution through the food chain begins with 156 .1 Origins and distribution It is recognised that several species of benthic dinoflagellates can produce CTXs and the gambiertoxins. These include species belonging to the genera Ostreopsis and Coolia. lipid soluble polyethers. and only a few structures have been identified.8.8). Culprit species producing CTXs may simultaneously produce unrelated toxins such as maitotoxin. Fig. although the dinoflagellate Gambierdiscus toxicus is considered to be the major source of these toxins. Such microalgae contribute to the primary production of tropical and sub-tropical reefs and live in epiphytic association with bushy red.
A good knowledge and careful observation of the symptoms in the bioassay (Minimal 157 .8. barracudas. but fish toxicity can vary widely depending on where the fish are caught and how long they have been feeding in that area. which in turn are consumed by carnivores such as snappers. and temperature sensation reversal. Like the brevetoxins. and inducing membrane depolarisation resulting in prolonged symptoms indicative of damage to nerve tissue. Patients have reported symptoms lasting weeks and even months. which can be confused with chronic fatigue syndrome. pruritus. Even when suspect samples are available. The recommended method to extract fish tissue has been described in detail (74) and the initial warm acetone extract is partitioned against hexane and diethyl ether where the CTXs are concentrated. the absence of a strong chromophore means that simple detection methods based on UV or fluorescence-based detection methods are ineffective. myalgia. and in fact CTX poisoning is often only identified after consumption of contaminated fish. and the sensation of loose teeth (77). Other symptoms include a metallic taste. The pharmacological effects following CTX exposure are numerous and include severe gastrointestinal distress and neurological symptoms. Local island populations attempt to avoid poisoning by catching only smaller fish. Another symptom that can develop over time is extreme fatigue. Globally. and it is estimated that ingestion of as little as 0.2 Toxicology Ciguatoxins (and maitotoxin which is often co-produced with CTXs) are extremely poisonous. 78). arthralgia. all of which can be linked to the effect of CTX on excitable membranes. 7. Differences in potency among the CTXs correlate directly with their binding affinity for the sodium channel.76). they bind to voltage-dependant sodium channels in the cell membrane causing them to open. and moray eels. and there are suggestions that such incidents are increasing and spreading worldwide (74). These include paresthesias (numbness and tingling of the extremities).8. the neurological symptoms can persist for days. sharks. weeks or even months (74 . though the mouse bioassay continues to be relied upon as the most common method of detecting CTXs. The onset of symptoms can occur rapidly within 24 hours. 75). where cold objects feel hot and vice versa. 75.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. Nevertheless CTX poisoning continues to be a threat to local and visiting populations.3 Detection methods As a consequence of their potency. 7. only trace amounts of CTXs can cause illness following consumption of contaminated fish. although not all CTXs are equipotent. there are neither standards nor an official testing program.1 µg of ciguatoxin can result in human illness (74. groupers. which can return following the consumption of alcohol and certain foods such as oily fish (74.
following consumption of contaminated mussels Mytelius edulis (82). but this only has application in a clinical setting (81). Chemical detection methods are severely compromised by the lack of standards. and only a few structures (AZA 1 . with AZA2 (10 .FISH AND SEAFOOD Lethal Dose (MLD) 0. and the separation of these polyethers from the lipid-soluble fraction of fish provides an additional challenge. at C-3 and C-23).g. minor revisions of a few stereocenters were made. and the corrected structure was published in 2004 (86) (Figure 7. Usually the major AZA derivatives found in shellfish are AZA1 (50 . Since the original discovery and identification of the toxin. 7.30%) and AZA3 (5 . and following synthesis of the molecule some time later. In this way Caribbean and Pacific CTX-1 compounds can be distinguished (74). and it is likely that most of these compounds are biotransformation products formed in the shellfish tissue. specific for CTX activity. The original structure proposed revealed an entirely new class of shellfish toxin.9 Azaspiracids Azaspiracid (AZA1) was originally identified in 1995 as the causative agent for a poisoning episode in Ireland. almost 20 additional derivatives have been reported (83 .5) have been fully characterised by Nuclear Magnetic Resonance (NMR) and MS analysis. 84).80%). which are based on an LC-MS/MS approach. A sodium channel-specific assay using mouse neuroblastoma cells has been described although this approach has still to be developed in a routine. Usually the additional AZA derivatives found in shellfish tissue are hydroxylated derivatives (e.85). 158 .25 µg/kg) can provide useful clues as to the toxins present but more specific and informative assays are required.20%) present in lesser amounts (Figure 7. The majority of these have been proposed based on LC/MS analytical data. Consequently. although specificity and sensitivity remain major hurdles (74) and the presence of DSP toxins affects the test (80). which features characteristic fragment ions corresponding to sequential losses of three molecules of water. has been developed. This is then subjected to LCMS/MS analysis. Immunoassays may be an alternative detection method (79). high-throughput screen (74). another in vitro sodium channel-based assay. Recently.9). and another dealing with the chemistry and detection methods has been published (85). Recent reviews of this new group of toxins have appeared (83.9). only a few laboratories are able to undertake such analyses. followed by clean-up on silica and aminopropyl cartridges. Acetone extracts of fish tissue are subjected to solvent partitioning.
Ostrea edulis).FISH AND SHELLFISH TOXINS Fig. and to date no long-term effects have been recorded (83.9. 7. Following the original episode.). cassipes (83). 89). which are ingested by P. 90). Structure of the three principal azaspiracids found in shellfish 7. cockles (Cerastoderma edule). Mytilus galloprovincialis). oysters (Crassostrea gigas. as in other cases of shellfish toxins. and most recently in a crustacean (Cancer pagurus). but current data indicates that unlike the DSP compounds. While this result is not necessarily in debate. which can persist for 2 . diarrhoea and stomach cramps. There has been considerable debate over the source of azaspiracids. AZA occurrence has been reported by several European countries including France. it has been impossible to establish that P. Donax spp.3 days. In fact the routes of trophic transfer of AZAs through the food chain ending in contaminated shellfish remain unknown. However.2 Toxicology Consumption of AZA contaminated shellfish can result in acute symptoms that include nausea. clams (Tapes philipinarum. and off the coast of Morocco (83. 7. 88. these molluscs were never considered to be the biogenetic source.9. Scandinavia. and some time after the original episode in Ireland. vomiting. Ensis siliqua. it was reported that the heterotrophic dinoflagellate Protoperidinium cassipes was the planktonic source of the toxins (87).9. scallops (Pecten maximus).1 Origins and distribution The azaspiracids were first isolated from contaminated shellfish. These symptoms are not unlike those experienced following the consumption of DSP toxins. and variable field results have led to suggestions that the actual producing organisms are Dinophysis spp. Portugal. cassipes produces AZAs in culture. AZA toxins do 159 . 84. and to date this has been the only source from which these toxins have been found at toxic levels. The bivalves affected include mussels (Mytilus edulis.
92). abnormalities in the thymus and spleen. This common cyclic imine feature is believed to be a common pharmacophore of this group of toxins which comprises the gymnodimines.3 Detection methods The regulatory level for AZAs in shellfish has been set by the EU at 160 µg AZA equivalents/kg shellfish tissue. although autopsies conducted after 4 and 8 hours revealed various gastrointestinal (GI) abnormalities including accumulation of fluid in the ileum. it is suggested that the order of toxicity is AZA2 (110 µg/kg) >AZA3 (140 µg/kg) >AZA (150 µg/kg). it is recommended that the entire shellfish be used for AZA analysis since it has been shown that these toxins can migrate to other tissues. the 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. and ASP. Using the same approach. Key fragment ions are useful in identifying the various AZA compounds that can occur in shellfish tissue (83. However. Due to their relatively recent discovery and limited knowledge of their toxicological properties and impact on human 160 . 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. PSP. The limited amounts of AZA toxins available continue to hinder a complete understanding of their mechanism of action. 91. though the possibility exists that they may inhibit other phosphatases. which are particularly amenable to detection by positive ion electrospray mass spectrometry.12). 7.FISH AND SEAFOOD not inhibit the phosphatases PP1 and PP2. Currently LC/MS and LC-MS/MS analysis is the preferred chemical analytical detection method for AZAs. 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). the spirolides. but unlike DSP monitoring methods which use only the hepatopancreas of the shellfish for extraction. Injection of a lethal dose of AZA1 in mice (>150 µg/kg) caused vacuole formation and fatty acid accumulation in hepatocytes. but all contain a spirocyclic or cyclic imine function within their structure. AZAs also display cytotoxicity towards various mammalian cell lines (83. 7. Importantly these pathological outcomes were reported as being different from those caused by DSP. 85). parenchymal cell pyknosis in the pancreas. The mouse bioassay has been used to monitor shellfish extracts. Once again. Based on IP minimum lethal dose data. and closely related ptierotoxins (Figures 7.10 – 7. and necrosis of epithelial cells on the microvilli (91). it is believed that the hydroxylated derivatives AZA4 (470 mg/kg) and AZA5 (<1000 mg/kg) are considerably less toxic.9. and erosion and bleeding in the stomach. the pinnatoxins.10 Cyclic imines These toxins are represented by several distinct structural types.
10. selliformis (97). it was proposed that a dinoflagellate identified as Gymnodinium spp. two additional congeners. 7.. Structure of the cyclic imine gymnodimine 7.10). surf clam and New Zealand abalone. cockles. 95). Some time later. 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. Fig.520 Da (94. a species closely related to Gymnodinium mikimoitoi was the planktonic source of the toxin. Subsequent taxonomic studies revised this again to a new dinoflagellate K. 161 .FISH AND SHELLFISH TOXINS health. Pacific oyster. 7.10. 95).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. gymnodimine B and C were isolated from the dinoflagellate Karenia selliformis (formerly Gymnodinium selliformis). In only two cases (gymnodimines and spirolides) has a planktonic source been identified. At the time of the initial observation of gymnodimine. The gymnodimines are the smallest members of the cyclic imine group with a molecular weight range of 500 . The structure is novel in that it contains a spirocylic imine group and a single tetrahydrofuran ring embedded within a macrocyclic structure. they are sometimes referred to as emerging toxins of uncertain human health concern.1 Gymnodimines The observation of neurotoxic symptoms in the mouse bioassay of lipophilic extracts of dredge oysters (Tiostrea chilensis) collected from around the South Island of New Zealand.1. led to the identification of a new marine toxin with an unprecedented structure named gymnodimine (96) (Figure 7. but they are included here for completeness. scallops.10. Reviews of the current status of the cyclic imine group were published by Molgo and Cembella (94.
Based on this diminished oral toxicity.FISH AND SEAFOOD 7. paralysis of the hind quarters and eventually complete immobility. finishing with 100% methanol.900 µg/kg). Most cyclic imines lack an extended chromophore.11).3 Detection methods The characteristic “fast-acting. 95) (Figure 7. Fractions are then dried and re-suspended in methanol or acetonitrile in preparation for LC/MS analysis (94. 95). Whether isolated from dinoflagellate cells directly or from shellfish tissue. 50%) of 96 µg/kg.to10-fold reduction of toxicity (LD50 600 .1. This led to the suggestion that gymnodimine acts by blocking nicotinic acetylcholine receptors at the neuromuscular junction. and there was a further ten-fold reduction in toxicity when the toxin was supplied with food. In particular. abdominal breathing. However. all or nothing” mouse bioassay response to gymnodimines and cyclic imines in general continues to be a useful monitoring approach.10. extracts exhibited the fast acting symptoms that became associated with cyclic imines following IP injection.10. Since their original discovery which resulted in the characterisation of spirolides A-F (99. edulis) and scallops (Placopectin magellanicus) from eastern Canada that displayed abnormal symptoms in the mouse bioassay. 7. and so spectroscopic detection by UV methods is not favoured. cyclic imines are readily observed by electrospray mass spectrometry in the positive ion mode. A later study using purified toxin established an LD50 dose (Lethal Dose. They were originally discovered in extracts of mussels (M. and short-acting cholinesterase inhibitors such as neostigmine or physostigmine protected mice against the toxic effects of gymnodimine. The time lapse between ingestion and death is between 5 and 15 minutes. 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.1. The original observation of toxicity in New Zealand followed from IP injection of lipophilic extracts in the mouse bioassay. The structure of the spirolides bears a strong resemblance to both gymnodimine and 162 . Reduction of the imine group eradicated the toxicity. it has been stated that regulation against gymnodimine is not required (98). There is a rapid onset of symptoms including respiratory distress. almost a dozen derivatives in total have been identified to date (94.100). 7. Oral administration of the toxin to mice by gavage resulted in about an 8.10.2 Spirolides The spirolides comprise the largest group of cyclic imine compounds. and this is the detection method of choice. together with a lack of evidence of human intoxication.2 Toxicology A review of the toxicology of all cyclic imines has been published (98). and lack of responsiveness to stimuli.
fasted mice).1 Origins and distribution At the time of the original discovery of the spirolides in shellfish from the coastal waters of eastern Nova Scotia. southern Ireland. They have also been reported in the southern hemisphere in Chilean mussels and in New Zealand shellfish and plankton (94. in fjords in Denmark and Norway. 7.11. fasted mice). the western coast of Scotland. 95). or by oral ingestion (381 .2. 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.707 µg/kg. While oral toxicity is considerably less than by IP injection. Fig.2 Toxicology The spirolides are the most studied group of the cyclic imines. those spirolides possessing the vicinal dimethyl grouping (see Figure 7. The LD50 values vary considerably depending upon whether the compound is administered by IP injection (5 . Interestingly. R1 = CH3) of the cyclic imine ring are the most toxic of the group (98).166 µg/kg. Since then the spirolides have been found to be globally distributed and are found in shellfish or plankton fractions along eastern Canada.10.FISH AND SHELLFISH TOXINS the pinnatoxins (vide infra).10. Structure of four spirolide toxins A-D 7. the gulf of Maine. their localisation in the digestive gland of mussels and scallops led to the suspicion that plankton were the real source of the toxins. This proved to be the case and eventually the dinoflagellate Alexandrium ostenfeldii was identified as the culprit organism (101).2.11. by gavage (87 . All data reported here are for desmethyl spirolide C.8 µg/kg). these are the toxins that are also most resistant to acid hydrolysis of the imine function 163 . 7. the Bay of Biscay and most recently from Mediterranean shellfish and plankton in the Adriatic (102).
12. 7. the mouse bioassay can be used to detect these fast acting toxins.B contain a spiro imine moiety similar to that found in the spirolides.12).10. Fig. 7. which includes two diastereomers. 7. Three members of this group have been reported.2.3 Pinnatoxins and pteriatoxins Pinnatoxins A . consistent with modulation of acetyl cholinergic receptors (103). more useful information on the amount and distribution of toxins present in a shellfish extract is obtained by LC-MS analysis usually following a simple clean-up step. While the mechanism of action remains to be clearly defined. 102).FISH AND SEAFOOD (100). four members of the group have been identified – two of them being the C-34 diastereomers. 95.10. experiments have shown upregulation of muscarinic and nicotinic receptors in mice following dosing with spirolide C. Structure of the pinnatoxin group and the closely related pteriatoxins 164 . The pteriatoxins were isolated from a different mollusc but bear a strong structural resemblance to the pinnatoxins (Figure 7.3 Detection methods As discussed for gymnodimine. To date. and in fact these compounds share a number of other structural features. Once again this relies on the sensitivity of these cyclic imines to detection by positive ion electrospray mass spectrometry. The compounds are zwitterionic and therefore somewhat more polar than the spirolides. However. A number of methods following this procedure have been described (94.
there is no compelling evidence to suggest that they have resulted in human toxicity (98).1 Origins and distribution Shellfish of the genus Pinna. However. 7. have long been suspected of causing human illness after ingestion. but this remains to be conclusively established (106). like the gymnodimines and spirolides. This prompted a chemical investigation of Pinna muricata that ultimately led to the identification of the pinnatoxins in 1995 (104).3 Detection methods No formal detection methods for these compounds have been reported. 7.3. which occupy tropical waters around Okinawa and the south east coast of China. This is generally a result of increased monitoring and continually improving detection technologies and methods. Despite this progress. new toxin groups such as the azaspiracids and cyclic imines have also been reported. Coupled with this. Only the acute toxicity in mice following IP injection has been reported. but the mouse bioassay data and LC-MS can be used to monitor suspect seafood for these compounds. once again reflecting improvements in purification and spectroscopic methods permitting structural identification of new and complex structures. There is strong circumstantial evidence to suspect that these toxins. 7. 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.10.10.FISH AND SHELLFISH TOXINS 7. and in some cases it is known that one toxin can potentiate or augment the toxicity of the other.11 Conclusion There has been considerable progress in the detection of known marine toxins. With increasing knowledge of the chemistry of toxins.100 µg/kg). The LD99 dose for the pteriatoxins is reported to be in the range (8 .10. 99%) values for pinnatoxins A-C are in the range (22 .3. which in many cases are now found to be more widespread than previously believed.180 µg/kg).most likely a dinoflagellate.2 Toxicology There is little information describing the toxicology of these compounds (98). while pinnatoxin D was essentially inactive. The LD99 (Lethal Dose. and later the closely related pteriatoxins were isolated from the digestive glands of another Okinawan bivalve Pteria penguin (105). While this is recognised as a real possibility for a wide range of toxins. and more advanced means of detection.3. arise from an algal source . has come the realisation that a “toxin profile” can be a complex mixture of many different classes of toxins. to date action or 165 .
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Boca Raton. 479-516. and detection.. Clark R.. Ciguatera fish poisoning in industrial ship crew members: a retrospective study in a seaport general practice in Trinidad and Tobago. Zaias J. 551-8. Cheng Y. Pierce R. Jacocks H. Ed. CRC Press.. Gandini C. 110.C.. Pierce R.. Toxicon. 187-94.. 2008. 68.. Baden D. Kirkpatrick B. Squicciarini D. CRC Press. 74... Pierce R.C.. 110. 220-6. Watkins S. Benson J. 179-85. Ramsdell J. 171 .. Food and Agriculture Organisation of the United Nations. Buonocore M. Lenzi A. 72. Poli M.. Granade R. 76.. 77.G. 117-71. 2004. 2007.. Poon-King T. Bourdelais A. 2004. Ciguatera fish poisoning: treatment prevention and management.D..R.M. Baden D... 73.S.. Naar J. Kubanek J.. The molecular and integrative basis to brevetoxin toxicity....M.. Backer L. in Seafood and Freshwater Toxins. Naar J. Food and Agriculture Organisation. 131. in Seafood and Freshwater Toxins. Chest.. 69. in Seafood and Freshwater Toxins. Environmental Health Perspectives. 99-115. Tomas C.. Wanner A. Flewelling L. Baden D.P.. 67. Environmental Health Perspectives..M. Tomas C. Fleming L. Backer L.P... Dickey R.E. Cheng Y.E. Muscle Nerve. Kirkpatrick B..S.S. Mild ciguatera poisoning: case reports with neurophysiological evaluations. Prockop L. Weisman R. Rome. Harmful Algae. Bean J.S.A. in FAO Food and Nutrition Paper 80... 2000.FISH AND SHELLFISH TOXINS 65.. Fleming L. Chen A.. 3. Botana L.D.. 2007. Marine Drugs. 456-79. Neurologic Shellfish Poisoning (NSP). Botana L.. Ciguatera toxins: Chemistry. Steidinger K. Ayyar R.. Whitney P.. 75.M. Aerosolised red tide toxins (Brevetoxins) and asthma. 465-70. Literature review of Florida red tide: implications for human health. Fernandez M.. Poon-King C. Granholm A. Baden D. 2008. CRC Press. 2008. Reich A... Flewelling L. 2007. Butera R.. Bossart G..B. Freidman M.. Ed.G..A. 707-23. Baden D. Bottein M-Y.G.. 1598-603.. Johnson D. Food and Agriculture Organisation. Fleming L.. 66. Naar J. Ed.G.. 2004..M. Abbott J. Brevetoxins. 70.. 825-34. Boca Raton. 2002. New fish-killing alga in coastal Delaware produces neurotoxins. Henry M.J. 71.W. Abraham W.. Dechraoui M-Y.. Locatelli C. Landsberg J. Ed.. Wang Z.. Lancaster J. Manzo L.N. Botana L. 23. 6.. Bourdelais A. 2002. toxicology. Detection of brevetoxins in the twenty-first century.. are potent ichthyotoxic neurotoxins that accumulate in fish. Marine Biotoxins. Schrank K. 519-50..E.. Abraham W. West Indian Medical Journal. Gannon D. Zaias J.. Kubanek J... Intrinsic potency for synthetically prepared brevetoxin cysteine metabolites BTX-B2 and desoxy-BTX-B2. 2008.P. Naar J. Competitive ELISA to detect brevetoxins from Karenia brevis (formerly Gymnodinium breve) in seawater. like ciguatoxins. Backer L..E.J. shellfish and mammalian body fluid. 53.G.H... Boca Raton. Ramsdell J.. 50. Wolny J.. 50. Dickey R. Toxicon.E.A. Bienfang P. Reich A.A.
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.
food microbiology.HACCP IN FISH AND SEAFOOD PRODUCT MANUFACTURE Step . from primary production to final consumption.3. Verification . 5.A point.Obtaining evidence that the elements of the HACCP plan are effective. 7. 8. quality assurance or control. Establish the corrective action to be taken when monitoring shows that a CCP is not under control. Establish the critical limit(s). Figure 8. 2. operation or stage in the food chain including raw materials. tests and other evaluations. hygiene managers.The application of methods. The team should include members familiar with all aspects of the production process as well as specialists with expertise in particular areas such as production. Each stage is discussed below in detail. food chemistry or engineering. procedure. 8.3 Stages of a HACCP Study The HACCP system consists of the following seven basic principles: 1. The team should also include 177 . Determine the CCPs. Conduct a hazard analysis. 3. Validation .1 Assemble HACCP team HACCP requires management commitment of resources to the process. in addition to monitoring to determine compliance with the HACCP plan. procedures. Establish a system to monitor control of the CCP. 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. 4. Establish documentation concerning all procedures and records appropriate to these principles and their application. An effective HACCP plan is best carried out as a multidisciplinary team exercise to ensure that the appropriate product-specific expertise is available. 6. ingredient and packaging buyers. Establish procedures for verification to confirm that the HACCP system is working effectively.1 is a flow diagram illustrating this 12-stage logic sequence.
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. If expert advice is not available on-site. biological.1.FISH AND SEAFOOD personnel who are involved with the variability and limitations of the operations. it may be obtained from external sources. Fig. 8.g. Logic sequence for application of HACCP 178 . chemical and/or physical).
These factors are likely to have a significant effect on some potential hazards. In addition. the only CCP that can render the product safe from pathogenic organisms is adequate heat treatment during preparation. is a cooking process required?). which should be described in detail.3.4 Construct a flow diagram The flow diagram should be constructed by the HACCP team and should contain sufficient technical data for the study to progress. shelf life.3. composition.). 8.1 Fish and seafood The HACCP team should ideally have access to expertise on the appropriate fisheries. fermentation.). contaminated fish and shellfish should not be used. It should provide an accurate representation of all steps in the production process from raw materials to the end179 . marine toxins associated with particular fish and shellfish species. such as children or the elderly. storage and distribution conditions and instructions for use.g. etc. The description should include information such as the product name. freezing. pH. etc. challenge testing and inoculation studies for evaluation of safety aspects are therefore essential for an effective HACCP team. physical and chemical structure (including water activity (aw). there may be species-specific hazards related to certain fish and shellfish. such as enteric viruses in shellfish. It is also important to identify the consumer target groups.g.3. However. including practices onboard fishing vessels. 8. for example contamination by foodborne pathogens as a result of time/temperature abuse or unsatisfactory handling practice. or in aquaculture operations. If the processing does not include a kill step. it must be noted here that certain toxins (algal and shellfish) are not destroyed by heat treatments. processing conditions (e.3 Identify intended use The intended use should be based on the expected uses of the product by the enduser or consumer (e. packaging.3. heat treatment.3.HACCP IN FISH AND SEAFOOD PRODUCT MANUFACTURE 8.2 Describe the product It is important to have a complete understanding of the product. Expertise on such inherent hazards. may need to be considered specifically. naturally present on fish from warm waters (> 15 °C).3. 8. aquaculture drug residues and potentially pathogenic Vibrio spp. 8. Vulnerable groups of the population.1.1 Fish and seafood Certain products may be contaminated or carry pathogenic organisms as a part of the natural flora.
Many fish-processing operations have relatively few steps and the flow diagrams appear simple. 180 ● ● . However. conduct a hazard analysis.3. 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.1 Fish and seafood Examples of flow diagrams for specific fish products may be found in the appropriate product chapters. 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. time/temperature data. adequate chilling of finfish immediately after capture is vital for the control of several potential hazards. features of equipment design. once determined. it is essential that the details of each step are fully appreciated and recorded.3.6 List all potential hazards associated with each step.4. It is also important to include observation of production outside normal working hours such as shift patterns and weekend working. It may include details of the factory and equipment layout.FISH AND SEAFOOD product. cleaning and hygiene procedures and storage conditions. survival or multiplication of pathogenic microorganisms. it should also include details of CCP steps. and any discrepancies between the flow diagram and normal practice must be recorded and the diagram amended accordingly. 8. It is essential that the diagram is accurate. because the hazard analysis and decisions regarding CCPs are based on these data. then any pre-drawn HACCP plans must be reviewed once the lines/products are finalised. Ideally.3. ingredient specifications. 8. 8. The operation should be observed at all stages. For example.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. If HACCP studies are applied to proposed new process lines or products. the qualitative and/or quantitative evaluation of the presence of hazards. and therefore a detailed description of the procedure applied must be available. or circumstances of any reclaim or rework activity. The analysis is likely to include consideration of: ● the likely occurrence of hazards and the severity of their adverse health effects.
3. Many of the microbiological hazards associated with fish and seafood products are derived from the raw materials.6. and the potential growth of Listeria monocytogenes on smoked fish. Pathogens may be part of the resident microflora of the living animal (e. to generalise about expected hazards and the reader is once again referred to the appropriate product chapter for additional advice on specific hazards. where a hazard is repeated.g. One control measure may be relevant to several process steps.g. The incidence of psychrotrophic. or desirable. any food safety objectives or manufacturer’s food safety requirements. and viruses). the number of consumers potentially exposed and their vulnerability. For example. Some hazards may require more than one control measure for adequate control and a single control measure may act to control more than one hazard. 8. Hazards introduced during processing of fish products depend very much on the characteristics of the process. since this may interfere with the analysis. the hurdle effect. non-proteolytic Clostridium botulinum on fresh fish is also sufficiently high that its presence may be assumed. or poor control of batter mixes in frozen battered fish portions may allow growth of Staphylococcus aureus and production of enterotoxin.). The HACCP team should then determine what control measures exist that can be applied for each hazard. Note: it is important at this stage that no attempt is made to identify CCPs. Therefore it is not possible. or may originate from polluted water or from post-capture contamination (e.1 Fish and seafood The term 'fish and seafood' includes an extremely varied group of products. 181 . 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. there are particular hazards associated with contamination of shellfish by microorganisms from human sewage. Hazards specific to certain types of product are detailed in the appropriate chapters of this manual. Vibrio spp. Salmonella spp. modified-atmosphere packaging (MAP) of chilled raw fish may provide conditions suitable for the growth of psychrotrophic C. For example. and there are an equally varied range of potential hazards associated with them.HACCP IN FISH AND SEAFOOD PRODUCT MANUFACTURE ● ● ● ● production or persistence of toxins.
1 Fish and seafood Specific product chapters provide information on criteria that may be used to set critical limits. 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. and its use may not always be appropriate. it can be said that effective control measures are likely to include the following: • • • • • 8. it is not possible to generalise on likely CCPs. application of the decision tree should be flexible. If a significant hazard has been identified at a step where control is required for safety. critical limits should be specified and validated for each CCP. 8. More than one critical limit may be defined for a single step.3. Where possible.FISH AND SEAFOOD 8. biological or sensory parameters.3.7 Determine CCPs The determination of CCPs in the HACCP system is facilitated by a decision tree (Figure 8.8 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.3. it is necessary to specify both time and temperature for a thermal process.1 Fish and seafood Again. However. It is also essential that the HACCP team has access to sufficient technical data to determine the CCPs effectively. then the process must be modified to include a control measure. chemical. given the enormous variety of fish products and processes in use. structured approach to decision making. and the reader is referred to the appropriate product chapter. 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 .7.2) to provide a logical. but for which no control exists at that step or any other. Criteria used to set critical limits must be measurable and may include physical. For example.8. However. 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.
14-9. WHO.L. 3 (1).HACCP IN FISH AND SEAFOOD PRODUCT MANUFACTURE Huss H. Bryan F. 1988.H. 1992. International Journal of Food Microbiology. International Commission on Microbiological Specifications for Foods. 15 (1/2). Hazard Analysis Critical Control Point Evaluations: A Guide to Identifying Hazards and Assessing Risks Associated with Food Preparation and Storage. Microorganisms in Foods. Food Control.. Volume 4: Application of the Hazard Analysis Critical Control Point (HACCP) System to Ensure Microbiological Safety and Quality. Development and use of the HACCP concept in fish processing. Mayes T. Simple users' guide to the hazard analysis critical control point concept for the control of food microbiological safety. 33-44. Oxford. 1992. Blackwell Scientific Publications. Geneva. 1992. World Health Organisation. 188 .
A number of standards are provided in food legislation. Hygiene is a general concept that covers a wide subject area. from structural conditions in the factory or process facility. to sale or supply to the final consumer.1 Introduction Hygiene is an important aspect of ensuring food safety. however. have been replaced by several linked measures on food safety rules and associated animal health controls. A new package of European Commission (EC) hygiene measures became applicable on 1 January 2006 to update and consolidate the earlier 17 hygiene directives. Microbiological standards have a useful role and help establish requirements for the microbiological safety and quality of food and raw materials.9. under which food is produced to optimise public health and to prevent. EC FOOD HYGIENE LEGISLATION Eugenia Choi and Dr. final product specifications. and conditions of raw materials. Procedures under the new legislation are based on risk assessment and management and follow a 'farm to fork' approach to food 189 . transport and delivery vehicles requirements. with the intention of introducing consistency and clarity throughout the food production chain from primary production. to personnel requirements. The general food hygiene Directive 93/43/EEC and other Directives on the hygiene of foodstuffs and the health conditions for the production and placing on the market of certain products of animal origin intended for human consumption. and one that plays an important role in most countries’ food legislation. including microbiological criteria. The new legislation was designed to establish conditions. the existence of microbiological standards alone cannot protect consumer health. Jenny Pfleger Leatherhead Food International Randalls Road Leatherhead Surrey KT22 7RY United Kingdom 9. 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. eliminate or acceptably control pathogen contamination of food.
handling and offering for sale or supply to the consumer. processing. 24.15). the following European Union (EU) hygiene regulations have applied: • • • Regulation (EC) No. N. transportation. 2073/2005 on microbiological criteria for foodstuffs • The general hygiene requirements for all food business operators are laid down in Regulation 852/2004. 190 • • .1992. 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. as last amended by Regulation (EC) No 806/2003. and sets out what those enforcing the provisions have to do. 7.1).9. 854/2004 of the European Parliament and of the Council laying down specific rules for the organisation of official controls on products of animal origin intended for human consumption Regulation (EC) No.1991. storing.2 Legislative Structure From 1 January 2006. J.1991. 9. Council Directive 92/48/EEC of 16 June 1992 laying down the minimum hygiene rules applicable to fishery products caught on board certain vessels in accordance with Article 3(1)(a)(i) of Directive 91/493/EEC (OJ L 187. 41).B. A number of more detailed implementing and transitional measures have been adopted at EC level. distribution. Regulation 854/2004 relates to the organisation of official controls on products of animal origin. Subsequently. Regulation 853/2004 supplements Regulation 852/2004 in that it lays down specific requirements for food businesses dealing with foods of animal origin. 852/2004 of the European Parliament and of the Council on the hygiene of foodstuffs Regulation (EC) No.FISH AND SEAFOOD safety with the inclusion of primary production in food hygiene legislation. p. European Communities L 268. as last amended by Regulation (EC) No 806/2003. 853/2004 of the European Parliament and of the Council laying down specific hygiene rules for food of animal origin Regulation (EC) No. Prescribed are detailed measures to ensure the safety and wholesomeness of food during preparation. packaging. Council Directive 91/493/EEC of 22 July 1991 laying down the health conditions for the production and the placing on the market of fishery products (OJ L 268. p.9. 24. manufacturing.7. p.
7.g. setting offences. 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 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.I. 2073/2005. revocation of existing implementing legislation.1993. 2006 No. Primary products are defined as products of primary production including products of the soil. The national legislation in all four UK countries also applied the provisions of the EU Microbiological Criteria Regulation No. the legislation is directly applicable law. p. food storage that is for private or domestic consumption.3 Regulation (EC) No. 19. 852/2004. penalties and powers of entry. 191 .) in England. with primary production. of stock farming. 9. domestic preparation or handling.11) The EU hygiene regulations apply to all stages of food production including primary production. There are some exemptions. This Regulation lays down general rules for food business operators on the hygiene of foodstuffs. Additionally the requirements of EC Regulation 853/2004 must be complied with. and equivalent legislation in Scotland. processing and distribution of food under their control satisfy the relevant hygiene requirements laid down in Regulation (EC) No. etc.LEGISLATION • Council Directive 93/43/EEC of 14 June 1993 on the hygiene of foodstuffs (OJ L 175.I. national legislation in the form of a Statutory Instrument (S. and also if the producer supplies small amounts of primary product to the final consumer or local retail establishments supplying the final consumer. 852/2004 on the General Hygiene of Foodstuffs Food business operators must ensure that all stages of production. e. as amended) came into force on 11 January 2006 (separate but similar national legislation also came into force on that day in Scotland. 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. right through to the implementation of procedures based on HACCP principles. and of hunting and fishing.14. and binding in its entirety on all member states from the date of entry into force. is required to give effect to the EU regulations. which will be covered later in this chapter. Wales and Northern Ireland). particularly taking into account a number of factors ranging from ensuring food safety throughout the food chain to begin with primary production. Wales and Northern Ireland. for example. Although the regulations have the force of law. As regulations. The Food Hygiene (England) Regulations 2006 (S. 1 .
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Annex I – Primary production
Annex I (Part A) relates to general hygiene provisions for primary production and associated operations covering: (a) the transport, storage and handling of primary products at the place of production, (b) the transport of live animals, (c) for fishery products - transport operations to deliver primary products (which have not been substantially altered) from the place of production to an establishment Food business operators have the responsibility to ensure primary products are protected against contamination. Any community and national legislation relating to the control of hazards in primary production such as measures to control contamination resulting from surroundings e.g. air, soil, water etc., and measures relating to animal health and welfare, and plant health that may impact on human health should be complied with. Requirements for record keeping are also laid down. This relates to animal feed (nature and origin), veterinary medicines administered to animals (date given and withdrawal periods), any diseases, analysis of samples from other animals that might impact on human health as well as reports on animal checks performed. Part B of Annex I contains recommendations for guides to good hygiene practice. 9.3.2 Annex II – Stages other than primary production
Annex II of the regulation lays down additional general hygiene requirements that must be met by food business operators carrying out production, processing and distribution of food at all stages following primary production. A summary of Chapters I to IV of Annex II is provided as follows: 220.127.116.11 Chapter I Chapter I applies to all food premises, except premises to which Chapter III applies. Food premises must be kept clean and maintained in good repair and condition. Their layout should allow for this. The environment should allow good hygiene practices and provide temperature-controlled handling and storage conditions where necessary ,and enable foods to be kept at correct temperatures and be monitored.
Additionally, there are requirements for provision of adequate lavatories, basins, ventilation, lighting and drainage.
18.104.22.168 Chapter II Chapter II applies to all rooms where food is prepared, treated or processed, except dining areas and premises to which Chapter III applies. The design and layout of rooms should allow for good hygiene practices between and during operations. Therefore floor and wall surfaces, ceilings and windows should be constructed to prevent dirt accumulating. Surfaces where food is handled must be well-maintained and allow easy cleaning and disinfection, preferably using smooth, washable corrosionresistant and non-toxic materials. There should be facilities for cleaning or disinfecting, and for storing working utensils or equipment. Clean potable water and adequate provision for washing food is needed.
22.214.171.124 Chapter III Chapter III applies to temporary premises (e.g. marquees, market stalls, and mobile sales vehicles), premises used primarily as a private dwelling-house but where foods are regularly prepared for placing on the market, and vending machines. Premises and vending machines should be practically sited, designed, constructed and kept clean and maintained in good repair and condition so as to avoid the risk of contamination, in particular by animals and pests. Facilities should allow adequate personal hygiene, and surfaces in contact with food should be easy to clean. Enough potable water and storage arrangements for hazardous or inedible substances are required as well as adherence to food safety requirements.
126.96.36.199 Chapter IV Chapter IV applies to all transportation. Conveyances and/or containers used for transporting foodstuffs are to be kept clean and maintained in good repair and condition to protect foodstuffs from contamination and are, where necessary, to be designed and constructed to permit adequate cleaning and/or disinfection. Food should be maintained at appropriate temperatures.
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188.8.131.52 Chapter V Chapter V refers to equipment requirements. Adequate cleaning and disinfection is to be carried out frequently for articles, fittings and equipment in contact with food where contamination needs to be avoided. Equipment should be installed to allow adequate cleaning, and be fitted with the required control device.
184.108.40.206 Chapter VI Chapter VI refers to food waste. Food waste, non-edible by-products and other refuse is to be removed from rooms where food is present as quickly as possible to avoid accumulation. Such waste is to be deposited in closable containers, to allow easy cleaning. Refuse stores should allow easy cleaning and be free of pests. Waste must be eliminated hygienically in accordance with European Community legislation.
220.127.116.11 Chapter VII Chapter VII refers to the water supply. There must be an adequate supply of potable water, which should be used whenever necessary to ensure foodstuffs are not contaminated. There are also requirements for recycled water, ice in contact with food, use of steam and for the water used in the cooling process for heat-treated foods in hermetically sealed containers. Specific requirements are that clean water may be used with whole fishery products. Clean seawater, may be used for live bivalve molluscs, echinoderms, tunicates and marine gastropods; and for external washing. If it is used, there must be adequate facilities to ensure the supply does not create a source of contamination for the foodstuff. Recycled water used in processing or as an ingredient must not present a contamination risk. It must be of the same standard as potable water, unless the competent authority is satisfied that the quality of water cannot affect the wholesomeness of food in the finished form.
Additionally, ice that comes into contact with food or which could possibly contaminate it needs to be made from potable water or, in the case of chilling whole fishery products using clean water. Ice must be made, handled and stored under conditions that protect it from contamination.
18.104.22.168 Chapter VIII Chapter VIII sets out personal hygiene requirements for those working in a food handling area including clean protective clothing and the requirement that those carrying or suffering from a disease are not permitted to handle food. 22.214.171.124 Chapter IX Chapter IX covers provisions applicable to foodstuffs. A food business operator should not accept raw materials or ingredients, other than live animals, or any other material used in processing products, if they are known to be contaminated with parasites, pathogenic microorganisms or foreign substances, to be toxic or decomposed to such an extent that, even after the business operator applied normal hygienic processing , the product would be inedible. Raw materials must be kept under appropriate conditions throughout production, processing and distribution. In particular, temperature control (i.e. cold chain and food thawing) requirements are laid down.
126.96.36.199 Chapter X Chapter X lays down provisions applicable to the wrapping and packaging of foodstuffs to avoid contamination of any form. 188.8.131.52 Chapter XI Chapter XI lays down heat treatment requirements for food that is placed on the market in hermetically sealed containers. The main relevant process parameters (particularly temperature, pressure, sealing and microbiology) must be checked by the food business operator to ensure that the required heat treatment has been achieved. Also, the process used should comply with an internationally recognised standard (e.g. pasteurisation, ultra high temperature or sterilisation). 184.108.40.206 Chapter XII Chapter XII states training requirements for food business operators to ensure that food handlers are trained in food hygiene matters and in the application of HACCP principles.
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The Regulation requires that food business operators must notify the relevant competent authority of each establishment under their control that carries out any of the stages of production, processing and distribution of food, with a view to the registration of each establishment. Food business operators must also ensure that the competent authority always has up-to-date information on establishments, including the notification of significant changes in activity, and closure of an existing establishment. Food business operators must ensure that establishments are approved by the competent authority, following at least one on-site visit, when approval is required by the national law of the Member State in which the establishment is located, or under Regulation (EC) No. 853/2004, or by a separate decision adopted. Separate rules apply for businesses producing products of animal origin. 9.3.4 HACCP
Food business operators, other than at the level of primary production, and associated operations must put in place, implement and maintain a permanent procedure or procedures based on principles of the system of hazard analysis and critical control points (HACCP). Emphasis is placed on risk-related control, with responsibility placed on the proprietor of the food business to ensure that potential hazards are identified and systems are developed to control them. Under HACCP, food business operators must amongst other considerations, identify hazards to be prevented or eliminated or reduced to acceptable levels, identify critical control points (CCP) at which control is essential to prevent, eliminate or reduce hazards and establish critical limits at these points, implement effective monitoring procedures, and establish corrective actions in the case where a CCP is out of control. Procedures must be followed to confirm the above is in place and up-todate, as well as providing the competent authority with documents and records as evidence, when required. 9.4 Regulation (EC) No. 853/2004 Laying Down Specific Hygiene Rules for Food of Animal Origin
Regulation (EC) No. 853/2004 lays down hygiene rules for products of animal origin that apply in addition to the general hygiene rules of Regulation (EC) No. 852/2004. 9.4.1 Definitions
The following definitions apply for fishery products and live bivalve molluscs:
purification centres or dispatch centres in tanks or any other installation containing clean seawater. shucking. mincing or processing. Factory vessel means any vessel on board which fishery products undergo one or more of the following operations followed by wrapping or packaging and. heading. slicing. where appropriate. after preparatory work such as bleeding. if necessary: chilling or freezing: filleting. live tunicates and live marine gastropods. 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. Prepared fishery products means unprocessed fishery products that have undergone an operation affecting their anatomical wholeness. reptiles and frogs) whether wild or farmed and including all edible forms. such as gutting. slime or sand. to remove mud. Freezer vessel means any vessel on board in which freezing of fishery products is carried out. slicing. where necessary. Bivalve molluscs means filter-feeding lamellibranch molluscs Marine biotoxins means poisonous substances accumulated by bivalve molluscs. gutting and removal of fins and. followed by wrapping or packaging. 197 . 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. Fresh fishery products means unprocessed fishery products. filleting. or in natural sites. whether whole or prepared. heading. 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. live echinoderms. and chopping. including products packaged under vacuum or in a modifiedatmosphere that have not undergone any treatment to ensure preservation other than chilling. Conditioning means the storage of live bivalve molluscs coming from class A production areas. in particular as a result of feeding on plankton containing toxins. shelling. and all mammals. skinning. parts and products of such animals.
For the UK. EG. EO. the food business operator may apply the same identification mark to both types of food. when a product's packaging and/or wrapping is removed or is further processed in another establishment. EK. 198 . the mark must show the approval number of the establishment.2 Fishery product requirements 9.FISH AND SEAFOOD 9. EK. EB or WE. Section I of Regulation (EC) 853/2004 are given as follows: 9.2.4. Section I of Regulation (EC) 853/2004. If an establishment manufactures both food to which Regulation 853/2004 applies and food to which it does not. a new mark must be applied to the product. (ii) However.1.2. 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.2 Form of the identification mark (i) The mark must be legible and indelible.4.1. (iii) Additionally. EC. EÜ. In this case. (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.4. Extracts of the requirements of Annex II.1 Identification marking An identification mark needs to be applied following the requirements of Annex II.2.4. 9. the code would be UK. ES. (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. (iv) If the identification mark is applied in an establishment within the Community.1 Application of the identification mark (i) The identification mark must be applied before the product leaves the establishment of production. the mark must be oval shaped and include the abbreviation CE. EY. the new mark must show the approval number of the establishment where this occurred. for other codes. and the characters easily decipherable and be clearly displayed. please refer to Regulation (EC) 853/2004.
the wrapping or the packaging. (iv) If products of animal origin are placed in a package destined for direct supply to the final consumer. C and D. (iii) In the case of liquid. it applies to those animals when not placed on the market live. including vessels. Chapter III. (v) When the mark is applied directly to products of animal origin. reference should be made to the actual Regulation. for full requirements. the approval number of the establishment and the abbreviation for the community where appropriate. This section does not apply to bivalve molluscs. The requirements of this section accompany those in Regulation (EC) 852/2004.e. an identification mark is not necessary if accompanying documentation contains the name of the country or its two-letter code.LEGISLATION 9. With the exception of Chapters I and II. and are intended for further handling. or be printed on a label affixed to the product. 9. Chapter IV. Section VIII of Regulation 853/2004. Extracts of the requirements of Annex III. i. (ii) For products of animal origin that are placed in transport containers or large packages.3 Method of marking (i) The mark is to be applied directly to the product. the colourings used must be EC authorised colours. processing.1. the requirements accompany those of Annex II of Regulation (EC) 852/2004 199 .2 Hygiene requirements for fishery products The regulation details specific hygiene requirements for fishery products. Part A and Chapter V apply to retail. Parts A. as amended. echinoderms. involved in primary production and such operations accompany the needs of Annex I of Regulation (EC) 852/2004 b) for other establishments including vessels.4. It may also be a tag made of a resistant material affixed in such a way that it cannot be removed. Therefore. granulate and powdered products of animal origin carried in bulk.2. the mark may be applied to the outer surface of the container or packaging. wrapping or packaging in another establishment.4. it is sufficient to apply the mark to the exterior of that package only. tunicates and marine gastropods when placed on the market live. and fishery products carried in bulk. specifically relating to fishery products are given below. its wrapping or the packaging. they would need to be obtained following Section VII on live bivalve molluscs.2. a) requirements for establishments.
4.2. heading. 2. the production of ice used to chill fishery products and the rapid cooling of crustaceans and molluscs after their cooking. i. The definition of fishery products covers primary production as well as its associated operations. they also include (i) the transport and storage of fishery products that have not been substantially altered (including live fishery products.FISH AND SEAFOOD c) for the water supply. in that record keeping requirements stated by point 7 in Part A of Annex I of Regulation 852/2004 do not apply to operators involved in small-scale coastal fishing (as defined through Regulation (EC) 1198/2006) if the activity is under 24 hours. (a) primary production covers the farming. it must be placed somewhere that will not contaminate the water supply. There is a derogation from point a. or to handle or process them after harvesting. fishing and collection of live fishery products with a view to their being placed on the market..1 Chapter I .2. refrigeration and wrapping. removing fins. surfaces that contact fishery products need to be made from cleanable corrosion-resistant material. Structural and equipment requirements refer to the following points 1. Requirements for vessels designed and equipped to preserve fresh fishery products for more than 24 hours 200 . from the place of production to the first establishment of destination. A.) 9. within fish farms on land) (ii) the transport of fishery products that have not been substantially altered (including live fishery products. Requirements for all vessels Vessels should be made in a way that does not cause contamination with substances such as fuel and oil etc. this supplements the requirements of Annex II. gutting.Requirements for vessels Food business operators are required to ensure that they meet the structural and equipment requirements as well as hygiene requirements required for vessels used to harvest fishery products from their natural environment. clean seawater may be used for the handling and washing of fishery products. and (b) related operations include any of the following operations if done on board fishing vessels: slaughter. Chapter VII of Regulation 852/2004.e. if a vessel has water coming in that is used with fishery products. bleeding.
(ii) Refrigeration equipment needs to keep fishery products in storage holds at not more than -18 °C. and not more than 0 °C after 16 hours. recording of temperatures. A temperature recording device must be present and placed where the temperature in the hold is the highest. Holds and containers used for storing fishery products must allow its preservation under adequate hygiene conditions and. (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.LEGISLATION (i) Such vessels need to have holds. any waste processing unit needs to have a separate hold to store the waste. 3. where necessary. (iv) There needs to be somewhere for storing packaging materials and this is to be separate from product preparation and processing areas. (iii) The above requirements for holds in vessels designed and equipped to preserve fresh fishery products for more than 24 hours must be met. 201 . Requirements for freezer vessels (i) These need to contain freezing equipment in order to achieve a core temperature of no more than -18 °C. tanks must incorporate devices that allow a uniform temperature throughout the tanks. (iii) In vessels equipped for chilling fishery products in cooled clean seawater. and allow the monitoring and. 4. 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. (v) Special equipment for disposing of waste/ inedible fishery products into the sea or a watertight tank and allocation of separate areas. where necessary. Also. 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. tanks or containers for the storage of fishery products at the temperatures laid down in Chapter VII. Storage areas for the finished products also need to be sufficiently large and cleanable. to ensure that melt water does not stay in contact with the products.
the product must be kept at no more than -9 °C. Handlers may use spiked instruments to move large fish or fish that might injure them. (ii) In the case of food business operators responsible for auction and wholesale markets etc. 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. do not have to meet the above requirements if no other form of handling or processing takes place on board such vessels. landed as soon as possible. B. d) Any gutting or heading of fish on board is to be done hygienically and quickly after capture. provided no damage occurs to the flesh. 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. c) Fishery products. must be chilled or. where fishery products are displayed for sale.4. Chapter II . b) Fishery products are to be handled and stored in a way that prevents the flesh bruising. e) In cases where whole fish intended for canning is frozen in brine. c) Factory vessels that freeze fishery products must have equipment meeting the requirements for freezer vessels as stated above.2. there need to be lockable facilities to refrigerate detained fishery products and separate lockable facilities to store fishery products that are unfit for human 202 . if this is not possible. and products washed thoroughly. chilled and wrapped. Parts that may present a danger to public health are to be removed quickly and kept away from products for human consumption.Requirements during and after landing 9. 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. other than those kept alive. Livers and roes for human consumption must be preserved under ice (temperature to approach that of melting ice) or be frozen.2.FISH AND SEAFOOD (vi) A water tank needs to be placed where there is no contamination of the water supply and hand washing taps should be designed to prevent contamination. b) Factory vessels on board which crustaceans and molluscs are cooked.
(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. 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 establishments. Chapter I. 9. handling fishery products A. there are requirements that such premises should be well lit and vehicles emitting fumes should not be able to access the premises. part I. including vessels. C. 854/2004. They should be wrapped. Gutting should ideally be done quickly once products have been caught or landed and afterwards be washed thoroughly. Requirements for mechanically separated fishery products (i) Raw materials used must satisfy the following 203 . Packaged fresh fishery products must be chilled to a temperature approaching that of melting ice. (iv) Filleting and cutting is to be carried out in a way that avoids contamination or spoilage of fillets or slices. C. dispatched. points 1 and 2. (ii) Heading and gutting must be carried out hygienically. packaged and chilled as quickly as possible after preparation.LEGISLATION consumption. and where necessary.3 Chapter III . 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.2. B. Fillets and slices must then be removed from work tables as soon as possible after preparation. Requirements for fresh fishery products (i) Where chilled or unpackaged products are not distributed.2.4. prepared or processed immediately after arrival at an establishment on land. (iii) Whole and gutted fresh fishery products can be transported and stored in cooled water on board vessels. 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. they must be stored under ice in appropriate facilities.
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. cooling must continue until a temperature 204 . or a stabilising treatment. Requirements concerning parasites (i) The fishery products below must be frozen at no more than . 9. except when supplied to the final consumer. mechanically separated fishery products must be frozen as quickly as possible or incorporated into a product intended for freezing. Requirements for cooking of crustaceans and molluscs After crustaceans and molluscs are cooked. (ii) mackerel. raw materials must have no guts.2. (iii) A document from the manufacturer.2.4 Chapter IV . (iv) (wild) Atlantic and Pacific salmon. (b) fishery products from the following species.Requirements for certain processed fishery products A. Once produced. stating the type of process they have undergone. (iii) sprat. they must be cooled rapidly. must accompany fishery products referred to above when placed on the market. if the processing is insufficient to destroy nematode larvae. (ii) The manufacturing process must satisfy the following Mechanical separation is to be carried out immediately after filleting. (ii) Compliance with the above is not necessary if epidemiological data indicates that the fishing grounds of origin do not present a health hazard in relation to the presence of parasites and this has been authorised by the competent authority.4. and whole fish must be gutted and washed beforehand.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.FISH AND SEAFOOD Only whole fish and bones after filleting may be used to produce mechanically separated fishery products. and (c) marinated and/or salted fishery products. If no other method of preservation is used. D.
processing. and after this. provided that the freshness criteria are met and the total volatile basic nitrogen (TVB-N) value of the unprocessed fishery products does not exceed the limits set out in point 1 of Chapter I of Section II of Annex II to Regulation (EC) No. pressing.5 Chapter V . (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. 9.2. 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.2. centrifugation. 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). raw materials need to be chilled as soon as possible and should meet the temperature requirements in Chapter VII.Health standards for fishery products In addition to compliance with microbiological criteria laid down in Regulation (EC) No. 2074/2005). However. and the raw material is processed within 36 hours after loading. cooked products are to be frozen or chilled to the temperatures stated in Chapter VII. the following conditions need to be met: A. Requirements for fish oil intended for human consumption (i) In relation to the raw materials used in their preparation and their transport. Histamine Histamine limits must not be exceeded (limits are set in Regulation (EC) No.4. C.LEGISLATION approaching that of melting ice is reached. 2073/2005. separation. 205 . refining and purification steps before being placed on the market for the final consumer. B. 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. Shelling/shucking is to be done hygienically. 2074/2005. B. 2073/2005).
prepared and processed fishery products belonging to the family Gempylidae. E. thawed unprocessed fishery products.2. and when fishery products are wrapped on board fishing vessels. it should be kept at no more than -9 °C. Diodontidae and Canthigasteridae.2. Fresh.6 Chapter VI .2.Storage of fishery products (i) Fresh fishery products. food business operators must ensure that wrapping material is not contaminated and is stored in such a way that it is not exposed to contamination and is easy to clean where necessary.2. The scientific name must accompany the common name on the label.4. and cooked and chilled products from crustaceans and molluscs are to be kept at a temperature close to that of melting ice. Fishery products contaminated with parasites may not be placed on the market for human consumption. 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. (ii) Frozen blocks prepared onboard vessels must be adequately wrapped before landing. 206 . 9. Toxins harmful to human health Fishery products derived from poisonous fish of the following families must not be placed on the market: Tetraodontidae. (ii) Frozen fishery products must be kept at a temperature of not more than -18 °C in all parts of the product.4.FISH AND SEAFOOD D. 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. Parasites Fishery products are to be visually examined so that visible parasites are identified before being placed on the market. 9.7 Chapter VII . in particular Ruvettus pretiosus and Lepidocybium flavobrunneum.Wrapping and packaging of fishery products (i) Containers in which fresh fishery products are kept under ice must be waterresistant and ensure that melt water does not remain in contact with the products. In the case of whole frozen fish in brine intended for the manufacture of canned food.
). 9.4. The temperature of frozen fishery products must not fluctuate above -15°C. (c) 35 mg of nitrogen/100 g of flesh for the species: Salmo salar. 2074/2005.2.3 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. Helicolenus dactylopterus. 9. provided that the journey is short and the competent authority allows this.2.LEGISLATION (iii) Fishery products kept alive are to be kept at a temperature which will not adversely affect food safety or its viability.4. (b) 30 mg of nitrogen/100 g of flesh for the species belonging to the Pleuronectidae family (with the exception of halibut: Hippoglossus spp. species belonging to the Gadidae family. melt water must not remain in contact with the products.8 Chapter VIII .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. Sebastichthys capensis. The following in relation to TVB-N limits is extracted from the regulation: Unprocessed fishery products shall be regarded as unfit for human consumption where organoleptic assessment has raised doubts as to their freshness and chemical checks reveal that the following TVB-N limits are exceeded: (a) 25 mg of nitrogen/100 g of flesh for the species: Sebastes spp. (ii) If fishery products are kept under ice. 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.2. species belonging to the Merlucciidae family. (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 . This covers Total volatile basic nitrogen (TVBN) limits and how to determine them.
FISH AND SEAFOOD Regulation (EC) No 853/2004.1 Chapter I . Extracts of the requirements of Annex III. please refer directly to the regulation. The reference method to be used for checking the TVB-N limits involves distilling an extract deproteinised by perchloric acid as set out in Chapter III For distillation apparatus and routine methods which may be used to check the TVB-N limits. VIII and IX. and point 3 of Chapter VII applies to retail. (b) In the case of other operations.General requirements for the placing on the market of live bivalve molluscs Live bivalve molluscs can only be put on the market for retail sale if done through a dispatch centre. reference should be made to the actual regulation. VI. The information required in the registration document is given in point 4 and includes the health status of the production area.4. Section VII of Regulation 853/2004.3. The copy of 208 . Chapters V. however. Apart from the provisions on purification. tunicates and marine gastropods. Chapter IX applies to pectinidae harvested outside those areas. for full requirements. The sample must consist of about 100 g of flesh. as amended. A food business operator can only accept batch of such bivalve molluscs provided correct documentary requirements are presented. 9. 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. shellfish species etc. they supplement the requirements of Annex II to that Regulation.3 The regulation details specific hygiene requirements for bivalve molluscs. taken from at least three different points and mixed together by grinding. The section is split into 9 chapters for which a summary is provided as follows: Chapters I to VIII apply to animals harvested from production areas that the competent authority has classified in accordance with Regulation (EC) No 854/2004. they supplement the requirements of Annex I to that Regulation. this Section also applies to live echinoderms. specifically relating to bivalve molluscs are given below.4. Member States may set limits at a higher level for certain species pending the establishment of specific Community legislation. Live bivalve mollusc requirements - 9. where the raw material complies with points (a). (b) and (c) of Part B(1) of that Chapter.
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. Requirements for production areas (i) The competent authority classes (classed A.Hygiene requirements for the production and harvesting of live bivalve molluscs A. C.2 Chapter II . 9. collection from: .LEGISLATION document must be kept for at least 12 months post dispatch or receipt or longer as stated by the competent authority.Class C means relaying over a long time before it can be placed on the market For bivalve molluscs not subject to Class B or C treatments.Class B means treatment in a purification centre or after relaying is done before it can be placed on the market . poles or other fixed means are to identify the boundaries of the sites.4. processing or relaying. 209 .Class A means they can be placed directly on the market . B or C) production areas with fixed locations and boundaries for where live bivalve molluscs may be harvested and this must be complied with. Other requirements such as conditions for relaying in order to allow optimal conditions for purification and for permanent record keeping are also specified. (ii) Provided the requirements of chapter V are met.3. Requirements for relaying live bivalve molluscs Only areas approved by the competent authority can be used for relaying live bivalve molluscs with a requirement that buoys. Transport needs to allow sufficient drainage for best survival conditions. Requirements for harvesting and handling following harvesting This includes requirements that any further handling should not cause contamination or excess damage to shells/tissues of the live bivalve molluscs that might affect their suitability for purification. B.
and there are requirements for tanks and water storage containers.Health standards for live bivalve molluscs In addition to microbiological criteria and organoleptic characteristic requirements. 160 micrograms of okadaic acid equivalents per kilogram.Structural requirements for dispatch and purification centres Premises on land should not be subject to flooding by tides etc.5 Chapter V .3.3. Requirements for dispatch centres Handling should not result in contamination to any degree. 1 milligram of yessotoxin equivalent per kilogram.4.3 Chapter III . the purification system to allow them to resume /maintain filter-feeding activity.4.3. the levels of marine biotoxins should not exceed the following limits. (iv) yessotoxins. 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. Before dispatch. containers and a label to identify purification.FISH AND SEAFOOD 9. 210 . requirements if there are many batches of live bivalve molluscs.Hygiene requirements for purification and dispatch centres A. B. dinophysistoxins and pectenotoxins together. 20 milligrams of domoic acid per kilogram (iii) okadaic acid. Requirements for purification centres There are requirements for live bivalve molluscs to be washed.4 Chapter IV . 800 micrograms per kilogram. 9. for: (i) paralytic shellfish poison (PSP). (ii) amnesic shellfish poison (ASP).4.
4. they cannot be re-immersed in or sprayed with water.3.LEGISLATION (v) azaspiracids.9 Chapter IX .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.2. must be waterproof. (ii) Once the live bivalve molluscs have been packed for retail sale and have left the dispatch centre. (Note: testing methods for detecting marine biotoxins are detailed in Annex III of Regulation (EC) 2074/2005) 9. the label. 9. the species of bivalve mollusc and date of packaging (at least the date and month) are required.7 Chapter VII .3.3.6 Chapter VI .4. 211 . (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. by way of derogation from directive 2000/13/EC. Registration document requirements also apply.1).Identification marking and labelling In addition to general identification marking (contained in 9.Wrapping and packaging of live bivalve molluscs (i) Oysters must be wrapped or packaged with the concave shell downwards. 9.4. Also. Requirements of Chapters III and IV are to be complied with.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. 9. The requirements for production areas as stated earlier apply to pectinidae and these can only be placed on the market through a fish auction.4.3. including the identification mark. the date of minimum durability may be replaced by the words ‘these animals must be alive when sold’.4. Finally.8 Chapter VIII . dispatch centre or processing establishment.
Chapter V of Regulation 853/2004 apply.FISH AND SEAFOOD 9. The official controls include audits of good hygiene practices and hazard analysis and critical control point (HACCP)-based procedures.5. (ii) fishery products not complying with the requirement concerning biotoxins. Therefore the health standards for fishery products as previously stated in Annex III. The requirements refer to hygienic conditions and regular inspections both on land and at port to ensure compliance of temperature and other requirements.1 Regulation (EC) 854/2004 Annex III Chapter III states that fishery products are to be declared unfit for human consumption if: organoleptic. or (iii) bivalve molluscs. 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. general principles for official controls in respect of all products of animal origin falling within the scope of the regulation are given.5 Regulation (EC) No. 854/2004 of the European Parliament and of the Council Laying Down Specific Rules for the Organisation of Official Controls on Products of Animal Origin Intended for Human Consumption Regulation (EC) 854/2004 gives requirements for official controls on products of animal origin and states requirements for those enforcing the provisions. or - - 212 . In this regulation. 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. 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. chemical. they derive from: (i) poisonous fish. Fishery products in particular must comply with the requirements of Annex III of the regulation. tunicates or marine gastropods containing marine biotoxins in total quantities exceeding the limits referred to in Regulation (EC) No 853/2004. echinoderms. 9. Section VIII.
live tunicates and live marine gastropods. - - - - B. The reference method for this analysis is the five-tube. Regulation (EC) 854/2004. three dilutions Most Probable Number (MPN) test specified in ISO 16649-3. Annex II 9.000 E. 000 E. that microbiological criteria and chemical contaminants amongst other requirements are met. section VII. Classification of production and relaying areas: This section reinforces that specified in Annex III. In the remaining 10 % of samples. three dilutions MPN test specified in ISO 16649-3. The reference method for this analysis is the five-tube.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. Sampling plans and frequency are required.2. live bivalve molluscs must not exceed 46. coli per 100 g of flesh and intravalvular liquid. as previously stated. 4.1 Chapter II . Also. live echinoderms. chapter II on hygiene requirements for the production and harvesting of live bivalve molluscs in Regulation (EC) 853/2004. Requirements for the competent authority are stated in relation to the classification or a production or relaying area is stated.2 This states official controls for the production and placing on the market of live bivalve molluscs. Live bivalve molluscs from class A areas may be collected for direct human consumption provided health standards are met.5. Monitoring of classified relaying and production areas This is to ensure there is no malpractice. 213 . Live bivalve molluscs from Class B areas must not exceed.5.Official controls concerning live bivalve molluscs from classified production areas A. Live bivalve molluscs from Class C areas must not exceed 46.600 E. coli per 100 g of flesh and intravalvular liquid. in 90% of the samples. chapter V is reinforced. coli per 100 g of flesh and intravalvular liquid. A summary is given as follows: 9.
Procedures for re-opening a closed production area are stated D. 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. which has applied since 1 January 2006.5.3 Imports By way of derogation from Article 11(1) of Regulation (EC) No 854/2004. 2073/2005 on Microbiological Criteria for Foodstuffs Regulation (EC) No.Official controls concerning pectinidae harvested outside classified production areas These are to be carried out in fish auctions. 9. establishes microbiological criteria for a range of foods. F. 9.2.5. 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. E. 214 . opening or closing a production area. Decisions after monitoring There are specific requirements whereby if sampling results indicate that health standards for molluscs are exceeded or there is a risk to human health. 9. the competent authority is to close the production area.FISH AND SEAFOOD C.6 Regulation (EC) No. 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.2 Chapter III . dispatch centres and processing establishments in order to confirm that health standards and other specific requirements are met. Food business operators' own checks Controls that food business operators have done are considered by the competent authority when classifying. boundaries or class of a production area or its closure. 2073/2005.
The definition of ‘microbiological criterion’ means a criterion defining the acceptability of a product.2 Process hygiene criteria Chapter 2 focuses on process hygiene criteria. 215 .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.4 referring to fishery products. ensuring that foods being placed on the market do not pose a risk to human health.LEGISLATION The aim of this legislation is to complement food hygiene requirements. 9. per unit(s) of mass.I. and the legislation applies to all businesses involved in food production and handling. and/or on the quantity of their toxins or metabolites. a batch of foodstuffs or a process. Two kinds of criteria have been established: food safety criteria.II.6. volume.6. 9. with chapter 2. based on the absence. The criteria are outlined in Table 9. area or batch. applying to products placed on the market and process hygiene criteria that are applied during the manufacturing process. The relevant criteria are outlined in Table 9. presence or number of microorganisms.
16 Cooked crustaceans and molluscan shellfish Salmonella EN/ISO 6579 Products placed on the market during their shelf life .2 Ready-to-eat foods able to support the growth of L. monocytogenes. 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. monocytogenes. 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.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 1.3 Ready-to-eat foods unable to support the growth of L. who has produced it EN/ISO 11290-2 (6) Products placed on the market during their shelf life 1.TABLE 9. monocytogenes 5 0 Absence in 25 g (7) FISH AND SEAFOOD 216 5 0 100 cfu/g 5 0 Absence in 25 g 1. monocytogenes L. other than those intended for infants and for special medical purposes(4) (8) L.
27 Fishery products which have undergone enzyme maturation treatment in brine.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. coli (15) 1. tunicates and gastropods 9 (18) 2 E.25 Live bivalve molluscs and live echinoderms.Food category Microorganisms Sampling plan(1) n 5 0 Absence in 25 g EN/ISO 6579 c m M Products placed on the market during their shelf life Products placed on the market during their shelf life Products placed on the market during their shelf life HPLC (19) Products placed on the market during their shelf life 400 g mg/kg Limit(2) Analytical reference method(3) Stage where the criterion applies 1.17 Live bivalve molluscs and live echinoderms. manufactured from fish species associated with a high amount of histidine (17) Histamine .
Relevance of matrix effect in determination of biogenic amines in 6 7 8 15 16 17 18 19 218 .0 and aw ≤ 0.94. J. monocytogenes. Pomatomidae. Particularly fish species of the families: Scombridae. — fresh.. soft drinks. when he is not able to demonstrate. including cocoa and chocolate products. References: 1. A pooled sample comprising a minimum of 10 individual animals.25. live bivalve molluscs. spirits and similar products. beer.. This criterion shall apply to products before they have left the immediate control of the producing food business operator. Duflos G. uncut and unprocessed vegetables and fruits. biscuits and similar products. — bread. Clupeidae. that the product will not exceed the limit 100 cfu/g throughout the shelf life. — — bottled or packed waters.4 or aw ≤ 0. 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. to the satisfaction of the competent authority. Single samples may be taken at retail level. Other categories of products can also belong to this category. 1996. 5 This criterion shall apply if the manufacturer is able to demonstrate. Products with pH ≤ 4. excluding sprouted seeds. shall not apply. products heat treated in their final package). 79. Malle P. cider.1. c = number of sample units giving values between m and M For point 1. m = M. Bouquelet S. 1 ml of inoculum is plated on a Petri dish of 140 mm diameter or on three Petri dishes of 90 mm diameter..1 . 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. 43-9. subject to scientific justification. 2. E. AOAC Internat. The most recent edition of the standard shall be used.. Bouquelet S. sugar. to the satisfaction of the competent authority. Engraulidae..92. wine. In such a case the presumption laid down in Article 14(6) of Regulation (EC) No 178/2002. 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. that the product will not exceed the limit of 100 cfu/g throughout the shelf life. Coryfenidae. Malle P. when recontamination is not possible after this treatment (for example. products with pH ≤ 5. according to which the whole batch is to be deemed unsafe. Scombresosidae. Assay of biogenic amines involved in fish decomposition. honey and confectionery. Dervin C. Valle M.FISH AND SEAFOOD 1 2 3 4 n = number of units comprising the sample.
J. — unsatisfactory. if all the values observed indicate the absence of the bacterium. if any of the values are > the limit. — unsatisfactory. AOAC Internat.LEGISLATION plaice (Pleuronectes platessa) and whiting (Merlangus merlangus. if the presence of the bacterium is detected in any of the sample units. the mean value observed is ≤ m 2. tunicates and gastropods in relation to testing E. coli. monocytogenes in ready-to-eat foods able to support the growth of L. if the presence of the bacterium is detected in any of the sample units.I The limits given refer to each sample unit tested. Interpretation of the test results relating to Table 9. where the limit refers to a pooled sample. 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. L. monocytogenes before the food has left the immediate control of the producing food business operator when he is not able to demonstrate that the product will not exceed the limit of 100 cfu/g throughout the shelf life: — satisfactory. no values observed exceed the limit of M. (1) 219 . The test results demonstrate the microbiological quality of the batch tested(1). L. L. — unsatisfactory. if all the values observed indicate the absence of the bacterium. The test results may be used also for demonstrating the effectiveness of the hazard analysis and critical control point principles or good hygiene procedure of the process. Histamine in fishery products from fish species associated with a high amount of histidine: — satisfactory. Salmonella in different food categories: — 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. 1097-101. monocytogenes in other ready-to-eat foods: — satisfactory. — unsatisfactory. 82. if the presence of the bacterium is detected in any of the sample units. 1999. if all the values observed are ≤ the limit. excluding live bivalve molluscs and live echinoderms. if the following requirements are fulfilled: 1. monocytogenes in ready-to-eat foods intended for infants and for special medical purposes: — satisfactory.
4. 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.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 2.1 Shelled and E.
Coagulase-positive staphylococci in shelled and cooked crustaceans and molluscan shellfish: — satisfactory. — unsatisfactory. if one or more of the values observed are > M or more than c/n values are between m and M. if a maximum of c/n values are between m and M. but there is also a general requirement that foods must not be kept at temperatures that would result in a risk to health. The regulations provide for defences in relation to upward variations of the 8 °C temperature. S. The defendant may be required to produce well-founded scientific proof to support his claims. if one or more of the values observed are > M or more than c/n values are between m and M. Schedule 4 of the Food Hygiene (England) Regulations 2006. coli in shelled and shucked products of cooked crustaceans and molluscan shellfish: — satisfactory. 9.7 Food Hygiene (England) Regulations 2006. S. and the rest of the values observed are ≤ m.LEGISLATION 1 n = number of units comprising the sample. if a maximum of c/n values are between m and M. tolerance periods for chill-holding of foods and hot-holding variations. if all the values observed are ≤ m. The test results demonstrate the microbiological quality of the process tested. — acceptable.1 In the U. E.K. 2006 No. the defendant will need to prove that the food was on service or display.II The limits given refer to each sample unit tested. with chill holding tolerance periods.I. — acceptable.I. The regulations prescribe a chilled food holding temperature of 8 °C or less. 2 Interpretation of the test results relating to Table 9. 14 details temperature control requirements for foods in general. For example. if all the values observed are ≤ m. and particularly that perishable foodstuffs must not be kept at above the maximum recommended storage temperature. — unsatisfactory. 14 (Hygiene requirements specific to the U. 2006 No.7. which overrides the 8 °C requirement. had not been 221 . and the rest of the values observed are ≤ m.K) Temperature control requirements 9. c = number of sample units giving values between m and M The most recent edition of the standard shall be used. Hot-held foods (food having been cooked or reheated and is for service or on display for sale) must not be kept below 63 °C.
preparing or processing the food recommending that it be kept at or below a specified temperature between 8 ºC and ambient temperatures. the period for which it can be expected to remain fit for sale when kept in a manner consistent with food safety.food. It should be noted that the temperature control requirements as detailed in Schedule 4 of the Food Hygiene (England) Regulations 2006 (S. For other food. it would need to be proved that the food was being transferred to or from a vehicle used for the activities of a food business. 2006 No.FISH AND SEAFOOD previously put on display at more than 8 °C and had been kept there for less than four hours. 2006 No. such as that below. was kept at above 8 °C or the recommended temperature for an unavoidable reason.S.gov.I. 9. 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.8 Guidance In the U. 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.K. Schedule 4 of the Food Hygiene (England) Regulations 2006 contains several definitions.I. to or from premises (including vehicles) at which the food was to be kept at or below 8 °C or the recommended temperature. including: Shelf life: where the minimum durability or 'use by' indication is required according to Regulation 20 or 21 of the Food Labelling Regulations 1996 (form of indication of minimum durability and form of indication of ‘use-by date’) the period up to and including that date. for a limited period consistent with food safety. S. The permitted reasons are given below: to facilitate handling during and after processing or preparation the defrosting of equipment. 3). or. 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. and was kept at above 8 °C or the recommended temperature. Alternatively. 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.uk/foodindustry/guidancenotes/hygguid/fhlguidance/ 222 . Recommended temperature: a specified temperature that has been recommended in accordance with a food business responsible for manufacturing.
1662/2006. L226. as read with the corrigenda at OJ No. 2074/2005.2004. L139. 854/2004 is now set out in a Corrigendum (OJ No. 1791/2006 and 1020/2008 and as read with EC Directive 2004/41. 30. laying down common marketing standards for preserved sardines. 853/2004 is now set out in a Corrigendum (OJ No.LEGISLATION 9. L226. 83) as amended by Regulation Nos 882/2004. these are not be discussed here. namely. and Regulation (EEC) 1536/92 laying down common marketing standards for preserved tuna and bonito. 2076/2005. 2074/2005.10 Further Reading Regulation (EC) No. 1688/2005.4.2004. L283. 854/2004 of the European Parliament and of the Council laying down specific rules for the organisation of official controls on products of animal origin intended for human consumption (OJ No. Regulation (EC) No. 30. 852/2004 of the European Parliament and of the Council on the hygiene of foodstuffs (OJ No.10.2005. 2075/2005 and Regulation (EC) No. L338. 1663/2006.2004.4. 2076/2005. 852/2004 is now set out in a Corrigendum (OJ No. Regulation (EEC) 2136/89. 55). L226. 22) as amended by Regulation (EC) Nos. Regulation(EC) No. 2074/2005. L139.2004.2004. as amended. 22. Regulation (EC) No. 223 .2004. 2076/2005. 25. 1019/2008 and as read with Regulation 2073/2005. 1. The revised text of Regulation (EC) No. Regulation (EC) No. The revised text of Regulation (EC) No. 853/2004 of the European Parliament and of the Council laying down specific hygiene rules for food of animal origin (OJ No. 206). 2076/2005. Regulation (EC) No.4.12. 62) as amended by Regulation (EC) No. 25.6. 2073/2005 on microbiological criteria for foodstuffs (OJ No. 9. 30.2006. L139.6. 2074/2005 and Regulation (EC) No. 1441/2007. 25. Regulation (EC) No.9 Other Relevant Legislation The following regulations about fish designations need to be considered: • Regulation (EC) No 104/2000 on the common organisation of the markets in fishery and aquaculture products Regulation (EC) No 2065/2001 laying down detailed rules for the application of Council Regulation (EC) No 104/2000 as regards informing consumers about fishery and aquaculture products • Note: Requirements for preserved fish are laid down elsewhere. The revised text of Regulation (EC) No. Commission Regulation (EC) No.6. 1791/2006 and 1021/2008 and as read with EC Directive 2004/41. 3) as amended by Regulation (EC) No. 1). 14.
83) Regulation (EC) No 104/2000 on the common organisation of the markets in fishery and aquaculture products (OJ L 17.12. 27 Commission Regulation (EC) No 2076/2005 laying down transitional arrangements for the implementation of Regulations (EC) No 853/2004. 22.2005. 22) Commission regulation (EC) No 2065/2001 laying down detailed rules for the application of Council Regulation (EC) No 104/2000 as regards informing consumers about fishery and aquaculture products as amended by Commission Regulation (EC) No 1792/2006 of 23 October 2006 (OJ L 278. 6) 224 . derogating from Regulation (EC) No 852/2004 of the European Parliament and of the Council and amending Regulations (EC) No 853/2004 and (EC) No 854/2004.1. 23.2001.10.FISH AND SEAFOOD Commission Regulation (EC) No 2074/2005 laying down implementing measures for certain products under Regulation (EC) No 853/2004 of the European Parliament and of the Council and for the organisation of official controls under Regulation (EC) No 854/2004 of the European Parliament and of the Council and Regulation (EC)No 882/2004 of the European Parliament and of the Council. (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 338.2000.12. as amended by Commission Regulation (EC) No 1022/2008 OJ L 338. 21. 22.2005.
1. PATHOGEN PROFILES 10.3 . 10.1.3.10. The minimum pH value for growth is thought to be pH 5.4 Heat resistance Aeromonads are sensitive to moderately high temperatures. and D-values at 45 and 51 °C have been reported as 29. under optimal incubation temperatures. Aeromonas has the potential to grow over a pH range of ca 4 . no growth is observed (1).5.1. respectively (1).1 Aeromonas spp. 225 .1.0 x 1.3 minutes.1.0 . Some death will occur at this pH if the temperature is 4 °C. 10.000-fold increase in number is observed after one week in food systems (1).10 (3). 0. At pH 4. At 5 °C. The optimum growth temperature is 28 °C (2). Morphology Gram-negative rods.2 Oxygen requirements Facultative anaerobe.5. a 10 to 1.5 pH Under otherwise optimal conditions.1.5 minutes.1 10. 10.45 °C (1). and 2.3 Temperature Aeromonas grows at temperatures of 2 .5 µm 10.
F are the types that are responsible for most cases of human botulism (5. forming at least seven different toxins. and is sensitive to > 4.1.2% sodium chloride (NaCl) (1).22.4 Heat resistance The vegetative cells of C.0 µm. botulinum types produce heat labile toxins.6 Aw/Sodium chloride Growth of Aeromonas is optimal in the presence of 1 . being capable of slow growth and toxin production at low temperatures . 10. Thus an aerobically packed product may not support the growth of the organism on the surface.2. but the minimum temperatures required to inhibit Groups I (proteolytic group) and II (non-proteolytic group) are different. at 85 ºC for 5 minutes. 6).even as low as 3 ºC (7).30 minutes. botulinum are known. botulinum grow reasonably well in the temperature range of 20 45 ºC. 226 . 10. or at 90 ºC for a few seconds. Types A.7 . All C. spore-forming rods. Group I will not grow at temperatures of 10 ºC or less. botulinum is an obligate anaerobe. to a lesser extent.2.2. and groups I and II are those responsible for the botulism cases.5 . 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.2 Clostridium botulinum Seven different types of C.2. types B and F can be either.5% NaCl (4).FISH AND SEAFOOD 10.3 Temperature All strains of C.2 Oxygen requirements Although C. 10. B.1 Morphology Gram-positive. A to G. All type A strains are proteolytic and type E strains are usually non-proteolytic. but the spores of this organism are more so. 0. E and. There are four main groupings of the organism.2. 10. 10. botulinum are not particularly heat resistant. botulinum is prevented. which may be inactivated by heating at 80 ºC for 20 . but Group II strains are psychrotrophic. many foods that are not obviously ‘anaerobic’ can provide adequate conditions for growth.4 x 1.
the minimum aw for growth may be lower. 10.10. therefore.21 minutes) (9).0.97 for Group II (5). perfringens .94 for Group I and 0. they may survive mild heat treatments (70 . The organism will grow well in the centre of meat or poultry dishes.3 10. The spores of Group II (non-proteolytic/psychotrophic) strains are less heat resistant than Group I strains.3 .6 Aw/ Sodium chloride The minimum water activity (aw) for growth of C. particularly by cooking.7 Characteristics of C.220.127.116.11 Clostridium perfringens Morphology Gram-positive spore-forming rods. 0.6 and pH 5. 10.is an obligate anaerobe.2 Oxygen requirements C. 227 . but under optimum growth conditions 10% (w/w) NaCl is required to prevent growth of Group I. 10. pH and temperature.5 pH The minimum pH for the growth of proteolytic and non-proteolytic strains is pH 4. These concentrations correspond to limiting aw of 0.0 and aw > 0. botulinum depends on solute. vacuum packed.PATHOGEN PROFILES 10. parcooked meals with pH value > 5. Note also that if humectants other than salt are used. where oxygen levels are reduced. safety margins must be introduced to allow for process variability. respectively (8. It will not. In commercial situations.97) (10. botulinum are produced by type A and proteolytic B strains (D121ºC=ca 0. 10.2. and 5% (w/w) NaCl is necessary to prevent growth of Group II organisms.85 ºC) and their ability to grow at refrigeration temperatures necessitates their control in foods capable of supporting their growth (e.9 x 2.2.like other clostridia . 9). These values have been established under carefully controlled laboratory conditions.0 µm. 11)).0 .3. However. grow on the surface of foods unless they are vacuum or gas-packed.g. botulinum spores The most heat resistant spores of Group I C.
3. although prior growth at high temperatures or the presence of fat in a food will result in increased heat resistance. pH.5 minutes (17). Spores. As in the case of other factors limiting the growth/survival of this organism.7 Characteristics of C. 10. depending on strain and other conditions. 13). lower concentrations may be effective in combination with other factors.93 and 0. perfringens. however. Under otherwise ideal conditions.FISH AND SEAFOOD 10. etc.3 Temperature The most significant characteristic of C. perfringens in roast beef at 60 ºC is 14. type of solute.6 Aw/Sodium chloride C. 10. 13). (It is unusual for spores to be formed in foods after the growth of this organism (14)). perfringens has the potential ability to grow within the temperature range 15 . 0 . In addition. Salt concentrations of 6 . the enterotoxin is not heat resistant . very limited growth may occur at pH values over the range pH ≤ 5 and ≥ 8. perfringens is not tolerant of low water activities.45 ºC.10 ºC.4 Heat resistance Exposure to a temperature of 60 ºC or more will result in the death of vegetative cells of C. the limits for aw are affected by temperature. While some growth can occur at 50 ºC.3. will survive greater extremes of pH (and aw) (12.8% inhibit growth of most C. At cold temperatures. 16). The lowest aw recorded to support the growth of C.it is destroyed by heating at 60 ºC for 10 minutes (14. death of the vegetative cells of this organism usually occurs rapidly above this temperature (12. Its optimum temperature for growth is 43 . perfringens appears to be 0.97 (glycerol and sucrose respectively) depending on the solute used to control aw (13. Some studies indicate that the presence of 3% NaCl delays growth of C. The D-value for C.50 ºC. and their heat resistance is also affected by the heating substrate. perfringens in vacuum packed beef (18).3. perfringens in relation to food safety is the organism’s ability to grow extremely rapidly at high temperatures. vegetative cells die rapidly (12).3. although C.3. perfringens can vary quite considerably in their heat resistance.3.5 pH C. perfringens spores The spores of C. Recorded heat 228 . perfringens strains. 15. 10. perfringens is not a tolerant organism with respect to pH. It grows best at pH values between 6 and 7 (the same pH as most meats). 10. 18).
growth at low temperatures can be very slow.2 has been suggested as the lowest working limit (22). In foods. and where acetic acid is used as an acidulant.at refrigeration temperatures down to.especially at refrigeration temperatures.0 minutes for heat resistant spores to 1. monocytogenes is not a particularly heat resistant organism. -0. monocytogenes is reported to be 45 ºC (20).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. it is not a sporeformer.2 Oxygen requirements Aerobe or microaerophilic. so can be destroyed by pasteurisation.4 Heat resistance L. 10. 229 .4. respectively (21).4.4 .4.3 Temperature Listeria monocytogenes is unusual amongst foodborne pathogens in that it is psychrotrophic. However. is between 30 and 37 ºC.4 ºC is probably a more likely minimum for growth in foods (19).5 . 10.19 minutes.64. Morphology Gram-positive short rods.0.4. Under ideal conditions.2. using hydrochloric acid as an acidulant).98 and 0. or even below 0 ºC. and temperature. Its optimum growth temperature.4 10.5 x 0. 10.23.1 Listeria spp.8 minutes for heat sensitive spores (12). monocytogenes in crawfish tail meat at 55. however.2.PATHOGEN PROFILES resistance values at 95 ºC (D-values) range from 17. 1. requiring days or weeks to reach maximum numbers.4. pH < 5.albeit slowly .3 .0 µm. D-values for L. 10. 60 and 65 ºC were reported as 10. the lowest limit for growth is likely to be considerably higher . 10. the organism is able to grow at pH values well below pH 5 (pH 4. It has been reported to have slightly greater heat resistance than certain other foodborne pathogens.3 is the lowest value where growth has been recorded.6 . 0. The upper temperature limit for the growth of L. however. being potentially capable of growing .
0.6 10.6 Aw/Sodium chloride L. or even grow.1 Plesiomonas Morphology Gram-negative.5 . but has been shown to be able to grow at an aw level of 0.97.5.8. 10.0 µm.3 Temperature Plesiomonas can grow in the temperature range of 8 . peritrichous flagella. 10.5 . at salt levels found in foods (10 .2 Oxygen requirements Facultative anaerobe.0 .6. 10. It grows best at aw of ≥ 0. the optimum being 30 ºC (23).FISH AND SEAFOOD 10. 10. 10.1. 25).5.5. 10.3 µm.12% NaCl or more). Morphology Gram-negative short rods. 230 .5. It is likely to survive.7 x 1. monocytogenes is quite tolerant of high NaCl/low water activities. 0.5 10.1 Salmonella spp.4 Heat resistance Pasteurisation at 60 ºC for 30 minutes has been reported as being effective in killing Plesiomonas shigelloides (24).5.5.83 (20).45 ºC.90.0 x 1 .3.5 pH Plesiomonas has a pH range for growth of pH 4.6 Aw/Sodium chloride Plesiomonas can grow well at NaCl concentrations up to 4% (24.5 (24. 10.4. 25). 0. The bacterium may survive for long periods at aw values as low as 0.8 .
Salmonellae are quite resistant to freezing.6. temperature and pH. a heat resistant organism. 10. the optimum being 35 . substrate. However.5 pH Salmonella has a pH range for growth of pH 3.3 Temperature Salmonellae can grow in the temperature range of 7 . although salt tolerance increases with increasing temperature (28).9. pasteurisation and equivalent treatments will destroy the organism under normal circumstances.6.6. 10.93 (as reported in dried meat and dehydrated soup).945. However.4% NaCl. Some death will occur at pH values of less than about 4.6. strains vary in their ability to withstand heating. 231 .5.5.5 ºC.6. 10. or possibly 0. depending on serotype. Salmonella has the potential to grow at aw levels as low as 0. some strains are able to grow at temperatures as low as 4 °C (26).20 times more heat resistant than the average strain of Salmonella at high aw (27). The optimal pH for Salmonella growth is between 6. under otherwise ideal conditions. Salmonella senftenberg 775W is about 10 . with a z-value of 4 .0. D60 ºC values normally range from about 1 to 10 minutes. and with an appropriate acid.4 Heat resistance Salmonella is not a spore-forming organism.5 . and appropriate heat treatments must be determined experimentally for low-aw foods.7.37 ºC. 10. high fat or low moisture levels (low aw) will reduce the effectiveness of heat treatments. therefore.8 .PATHOGEN PROFILES 10. Furthermore. Salmonellae are quite resistant to drying.2 Oxygen requirements Facultative anaerobe.6 Aw/Sodium chloride Where all other conditions are favourable. It is not. The growth of Salmonella is generally inhibited by the presence of 3 . depending on the type of acid and temperature.48 ºC. Growth is slow at temperatures below about 10 ºC.
7 . aureus than citric acid).37 ºC (29).7. It can survive well at low temperatures. aureus can grow within the temperature range 7 – 48. 0. however.5). with an optimum of 35 .3 (optimum 7.7.20 minutes. aw and temperature (sensitivity to acid increases with temperature).3 Temperature Under otherwise ideal conditions.7.9 µm diameter. Under most circumstances. The limits for toxin production are also narrower than for growth. 10. depending on aw. aureus viability.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. but may cause some cell damage (30). under otherwise ideal conditions (29. the D60 ºC value is 2 .7 10.7. In meat macerate.7.7. 232 .4 Heat resistance Heat resistance depends very much on the food type in which the organism is being heated (conditions relating to pH. 31). 10.1 Staphylococcus aureus Morphology Gram-positive cocci.0 .2 to 9.0. the organism is heat sensitive and will be destroyed by pasteurisation. NB: The growth of Staph.FISH AND SEAFOOD 10. 10. stressed cells can also be less tolerant of heating. etc. Staph.5 ºC. Freezing and thawing have little effect on Staph. 10. aureus is more limited under anaerobic than aerobic conditions. fat content. The following relate to limits for growth only. Most strains of staphylococci can grow within the pH range 4. aw.2 Oxygen requirements Facultative anaerobe.). As is the case with other bacteria.
an aw of 0.7. with a temperature range for growth of 10 . aureus can grow in 7 .86 is the generally recognised minimum in foods (32). especially vacuum packed foods (33) Temperature: pH: *Aw: Atmosphere: Heat Resistance: Enterotoxins are quite heat resistant. In general. 233 . Staph.5 . 10.>0.8 10.87 and > 0.0 . 10. aureus is unusual amongst food-poisoning organisms in its ability to tolerate low water activities.10% NaCl. and solute.7. 31) between 0.6 µm 10.8. * dependent on temperature.3 Temperature The optimum temperature for growth has been reported as 30 ºC. An effect of increasing salt concentration is to raise the minimum pH of growth.8.8 x 1. 10.43 ºC (35).2 Oxygen requirements Facultative anaerobe.9. However.7 Limits permitting toxin production 10 . atmosphere. It can grow over the aw range 0. 34).99 little or no toxin production in anaerobically packed foods.7. but certain strains can grow in 20% NaCl. strain.2 .PATHOGEN PROFILES 10.2.46 ºC (optimum between 35 and 40 ºC) (very little toxin is produced at the upper and lower extremes) (31) 5. pH.83 .8. In general. 0.6 Aw/Sodium chloride Staph.99 aerobically under otherwise optimal conditions.4 .1 Vibrio cholerae Morphology Gram-negative curved rods. heating at 100 °C for at least 30 minutes may be required to destroy unpurified toxin (31.5) (29. Staphylococci are more resistant to salt present in foods than other organisms.0.0 (optimum 7.
998.6 Aw/Sodium chloride V.8. cholerae in crab meat homogenate at 60 ºC is 2.6 (35. It can grow in the presence of 0. 10. cholerae is sensitive to heat at temperatures of >45 ºC.97 . but not acid conditions. 234 .5 pH This organism is tolerant of high pH values.9.984 (35). 38). 10.2 Oxygen requirements Facultative anaerobe. declining in number during storage. 10.9.9 10. cholerae may persist on frozen fish for >180 days (36). cholerae can grow over the aw range 0. it has a pH range for growth of pH 5 .9.4.9. 10.0. The D-value for V. respectively. with an optimum of 0.3 Temperature The minimum and maximum temperatures reported for growth in laboratory media. with an optimum of 37 ºC (39). 10.1 Vibrio parahaemolyticus Morphology Gram-negative curved rods. with optimum at 7. are 5 and 43 ºC. parahaemolyticus is easily destroyed by drying.4 Heat resistance V.1 .5 ºC). 37.8. V. under otherwise favourable conditions.6.8. and it is sensitive to refrigeration temperatures (0 .0% NaCl (35).FISH AND SEAFOOD Also. there is evidence that V.65 minutes (35). 10. but only moderately sensitive to freezing (40).
10.10. 10.11.2 Oxygen requirements Facultative anaerobe.9.6 .1 Morphology Gram-negative curved rods. 235 .6 (41). with an optimum pH of 7. 10. parahaemolyticus is unable to grow in the absence of salt.02 . parahaemolyticus will grow at aw levels as low as 0.10. with a temperature range for growth of 8 . but the optimum concentration is 2 . its upper and lower limits for growth are 8 (although some reports suggest 10) and 0.8. 41).6 Aw/Sodium chloride V.3 Temperature The optimum temperature for growth has been reported at 37 ºC.4% (18.104.22.1682.4 Heat resistance V.0. The optimum concentration of NaCl equates to aw of approximately 0.98 (35. The D-values at 55 ºC for clam and crab homogenate are 0. 10. The organism can survive for long periods at freezer temperatures (42). and are readily destroyed by cooking. depending on solute.5 minutes.43 ºC (42). 10. 41).10. V. respectively.29 minutes.5% NaCl. parahaemolyticus are not heat resistant. 10. vulnificus is heat sensitive. 10. and 2.PATHOGEN PROFILES 10. it has been reported that cooking oysters for 10 minutes at 50 ºC should ensure destruction of the organism (43).5 pH The pH range for growth is 4.4 Heat resistance V. respectively (39).10 Vibrio vulnificus 10.8 .
121-76.10. Growth and heat resistance of psychrotrophic Clostridium botulinum in relation to ‘sous vide’. 1995. Peck M. 1994. Ed. Fratamico P.W. Lund B. 2001. London. Aeromonas hydrophila group.D. International Commission on Microbiological Specifications for Foods.10.O. 1-27. Johnson E. 1989.M.D.R. in Foodborne Bacterial Pathogens. Doyle M.D..5 .. Beuchat L.P. 383-408. Dodds K. in Foodborne Pathogens. in Clostridium botulinum: Ecology and Control in Foods.L.W.W.0% NaCl. References International Commission on Microbiological Specifications for Foods... Marcel Dekker.P. 6 (1). Stelma G. 11. 1996. Clostridium botulinum. 5.. in Clostridium botulinum: Ecology and Control in Foods. Hauschild A. Marcel Dekker Inc. vulnificus grows over the range 0. 1993.. Diseases Caused by Bacteria. 2. Abeyta C. in Microorganisms in Foods.P. 357-67. Clostridium botulinum... Cliver D. Eds. Volume 5: Microbiological Specifications of Food Pathogens.W.K. Food Control. Volume 1: Diseases Caused by Bacteria. 8.N. Caister Academic Press. 2005.. 10.H. in International Handbook of Foodborne Pathogens. Principles of control. Hauschild A.. Cliver D. Dodds K.11 1.C. Aeromonas hydrophila.L. Foegeding P. Gaze J.. New York. Blackie. Microbiology and Molecular Biology. 1989. Eds.5.H. 97-131. 1994. Eds. Juneja V. Marcel Dekker. New York. 10. Austin J..H. Potential hazards associated with REPFEDS.H. Hauschild A. 279-303. Dodds K.O. Ed.E. New York. Aeromonas. New York.... Doyle M. Aeromonas species.D. Palumbo S. in Foodborne Bacterial Pathogens. Murrell K. 57-63. Bier J. 10.10. Clostridium botulinum. 4. with an optimum pH of 7.6 Aw/Sodium chloride V. Eds. with an optimum of 2.. Novak J. 1-19.FISH AND SEAFOOD 10.L. Gorman J. Bhunia A. in Food Microbiology: Fundamentals and Frontiers. New York. 111-89.M. Washington D. Murrell K. 5-19.H. Ed. 1993. 6.. Montville T. 9. 3. in Foodborne Disease Handbook. Notermans S.. ASM Press. New York. Marcel Dekker..5 pH The pH range for growth is 5 .5% (42). in Foodborne Disease Handbook.R. Eds. Miliotis M. Volume 1.J. 329-49. Kim J. Betts G..N. 236 .W. Marcel Dekker.. Doyle M. Eds.H. Kovacek K.8 (42). Clostridium botulinum and Clostridium perfringens. Hui Y. Marcel Dekker. Stelma G.. Smith J. Eds. 2003. Faris A.A. Hui Y.M. Marcel Dekker.R. 7. Gorham J. New York. Great Britain..L.
Wrigley D. in Foodborne Bacterial Pathogens.A. Volume 5: Microbiological Specifications of Food Pathogens. Evans M. Washington D. International Commission on Microbiological Specifications for Foods. in Foodborne Pathogens: An Illustrated Text. Listeria.. Juneja V. 141-82.D. Eds. 2006.H.A. London. Listeria monocytogenes.. London... 237 . 1989. Marcel Dekker. London. International Commission on Microbiological Specifications for Foods. Lancet. 1996. New York. 129-38. Evans M. in Foodborne Diseases. Marth E. Walker S.H.M. Kim C. Elsevier. London.J. 16. Eds. 24. Marcel Dekker.K.A. Wolfe Publishing Ltd. 191-243. International Commission on Microbiological Specifications for Foods.. Varnam A.R. Ed. 18. Volume 5: Microbiological Specifications of Food Pathogens.C.T. 351-82. Hui Y. Blackie.PATHOGEN PROFILES 12.. 14. Listeriosis and Food Safety. in Food Microbiology .C.P. Cliver D. in Food Microbiology: Fundamentals and Frontiers. Academic Press.. Volume 5: Microbiological Specifications of Food Pathogens. London. 33.H. McClane B. Banks J. International Commission on Microbiological Specifications for Foods. Foodborne disease due to Bacillus and Clostridium species. 735-42. London. Montville T. Clostridium perfringens. New York. 1994. 26.O. Marcel Dekker. 208-13. 23. Doyle M. Avian Disease. International Commission on Microbiological Specifications for Foods. 336 (8721). Johnson E.. Blackie.H.J. Labbe R.. in Foodborne Infection and Intoxication.R. 2001. Varnam A. Plesiomonas shigelloides. 1996. Archer P. Blackie.G.J. Ed. New York.O. 1990.An Introduction. Plesiomonas shigelloides.P. Ed. 13. 19. 1991. in Microorganisms in Foods. Matthews K. Listeria monocytogenes.V. Nagaraja K. Cliver D. 1989.. Clostridium perfringens food poisoning. Marcel Dekker.. Ed. 21..A.. Ryser E. 112-25. Montville T. 1990..R. 2007. Journal of Applied Bacteriology.. 1990. 201-8. Clostridium perfringens gastroenteritis. Rinke H.G.P. Eds. 68 (2).P. Eds. 2005. Clostridium perfringens. Murrell K. Plesiomonas. 311-25. 22. 157-62. 20. 17. 1989. Effect of time and temperature on growth of Salmonella enteritidis in experimentally inoculated eggs. 1996. Volume 1. Koburger J. Lund B.. in Microorganisms in Foods. Ed. in Foodborne Disease Handbook. 15. Riemann H. Eds. 25. in Microorganisms in Foods. Doyle M. Doyle M. Emery D. Washington D.. 137-64. Labbe R.G. Gorham J. Beuchat L.J.M. Growth of Listeria monocytogenes at refrigeration temperatures. Cliver D. Halvorson D. 133-67. ASM Press.A. Diseases Caused by Bacteria.. ASM Press. 229-40. Clostridium perfringens. 982-6. New York. International Commission on Microbiological Specifications for Foods.O. in Foodborne Bacterial Pathogens. Ed. Clostridium perfringens.
Bhunia A. Volume 5: Microbiological Specifications of Food Pathogens. Microbiological Specifications of Food Pathogens.. Golden D.. Wilkinson B.M. Marcel Dekker. Ed. New York. 1989. Griffiths M. London.M.. Springer Science.A.J. Stress Response and Resistance. 543-68. New York. Vibrio parahaemolyticus. 34. 33. 28. in Microorganisms in Foods.. 642. Jay J. Blackie. 31. Wymondham.. 39. 85-106.K. Staphylococcus aureus as a food pathogen: staphylococcal enterotoxins and stress response systems. Jay J. in Foodborne Bacterial Pathogens. On the resisting power of Vibrio cholerae and typhoid Bacillus against low temperatures. Loessner M. 1930.H. Tohyama Y. Volume 5.L. 1996. Gustafson J. 523. Vibrio cholerae. Cliver D. Experimental investigations concerning the prevention of epidemic diseases due to aquatic products (Fourth Report). 1989. in Understanding Pathogen Behaviour Virulence. in Modern Food Microbiology. Doyle M. Doyle M.O. Volume 5. 4 (13). 2005. Cliver D. 29. London. 2005. Lee-Wong A.FISH AND SEAFOOD 27. 30.62. 40.M.C. Blackie. 2005. Woodhead Publishing Ltd. Foodborne illness (Part 1): Staphylococcal (“Staph”) food poisoning. 1996. Food and Environmental Sanitation.J. 13 (11).P. Twedt R. London. 273-84. Staphylococcal gastroenteritis. Eds. in Microorganisms in Foods. International Commission on Microbiological Specifications for Foods.D. Microbiological Specifications of Food Pathogens. Fratamico P. Vibrio parahaemolyticus. Ed. 327-445.. International Commission on Microbiological Specifications for Foods. Academic Press. Golden D. Applied and Environmental Microbiology. Caister Academic Press. Academic Press. 38.S. Reed G.. Japanese Journal of Experimental Medicine. Eds. International Commission on Microbiological Specifications for Foods. Marcel Dekker.R. in Foodborne Disease. International Commission on Microbiological Specifications for Foods. London. Eds.P. Oliver J.. 1993. R15764. 414-25. International Commission on Microbiological Specifications for Foods. 238 . in Foodborne Pathogens.P.C.A. Riemann H. Loessner M. Crowcroft N. Cambridge. 545-66. Blackie. 37. Staphylococcal Food Poisoning. Ed.S. 1990. in Microorganisms in Foods. Ed. Staphylococcus aureus. D’Aoust J-Y.A. 1983. CDR Review. Cholera: current epidemiology. 1996. in Foodborne Infections and Intoxications. Distribution of Vibrio vulnificus and other lactose-fermenting vibrios in the marine environment. Staphylococcal intoxications. 8. in Foodborne Pathogens: Microbiology and Molecular Biology. 36. Cleland D. International Commission on Microbiological Specifications for Foods.S. 217-64. Dairy. Ed.M. 32. Salmonellae. New York. 331-57. Salmonella. Ed.O. Bergdoll M. 327-52. 426-35. Warner R.. Ed. Smith J. Stewart G. 2005. London. 985-98. Bergdoll M.. 45 (3). 1994. 35.
W. 436-9.. Cold storage and mild heat treatment as processing aids to reduce the numbers of Vibrio vulnificus in raw oysters. Jay J.PATHOGEN PROFILES 41. Blackie. Volume 5. in Microorganisms in Foods. Golden D. Journal of Food Protection.D.. 239 . Loessner M. Vibrio vulnificus. 42. 43.A. Cook D. 2005..J. 657-78.M. Yersinia. London. in Modern Food Microbiology. Ruple A. New York. International Commission on Microbiological Specifications for Foods. 1996. and Campylobacter species. Ed. 1992. Loessner M. International Commission on Microbiological Specifications for Foods.. Springer Science. 985-9.. 55 (12).M. Jay J.A. Microbiological Specifications of Food Pathogens.J. Foodborne gastroenteritis caused by Vibrio. Golden D.
FISH AND SEAFOOD 240 .
gov.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/ .email@example.com Institute of Fisheries Management (IFM) 22 Rushworth Avenue West Bridgford Nottingham NG2 7LF United Kingdom Tel: + 44 (0)115 9822317 Fax: + 44 (0)115 9826150 Email: firstname.lastname@example.org/ Department for Environment Food and Rural Affairs (Defra) Nobel House 17 Smith Square London SW1P 3JR United Kingdom Tel: + 44 (0) 207 2386000 Fax: + 44 (0) 207 2382188 241 Email: email@example.com Web site: www.gov.co.gsi.CONTACTS Addresses of Trade Associations and Professional Bodies Association of British Salted Fish Curers and Exporters c/o Cawoods (Fish Curers) Ltd Estate Rd No 6 South Humberside Industrial Estate Grimsby North East Lincolnshire DN31 2TG United Kingdom Tel: + 44 (0) 1472 342248 Fax: + 44 (0) 1472 353100 Email: firstname.lastname@example.org Web site: www.org.uk or v.gsi.gov.defra.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.scotland.uk Web site: www.org.
email@example.com Danmarks Fiskehandlere H. Andersens Boulevard 37.dk Web site: www.is Web site: www.difta.no Web site: www.org Web site: www.C. Northern Ireland Fisheries and Rural Policy Division 4th Floor North Dundonald House Stormont Estate Belfast BT4 3SB United Kingdom Tel: + 44 (0) 28 90524999/ 4991 Fax: + 44 (0) 28 90378323 Email: firstname.lastname@example.org Web site: www.net Irish Sea Fisheries Board PO Box 12 Crofton Road Dun Laoghaire Co.bim.fiskehandlerne.uk/index/fisheriesfarming-and-food.dardni.eapo.uk Web site: www.13 Postbox 6122 N-9291 Tromsø Norway Tel: + 47 77 629000 Fax: + 47 77 629100 Email: email@example.com.FISH AND SEAFOOD Department of Agriculture and Rural Development.is/ 242 . 1 1553 København V Denmark Tel: + 45 35 372023 Fax: + 45 35 372033 Email: Info@fiskehandlerne.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.com/ Danish Institute for Fisheries Technology and Aquaculture (DIFTA) illemoesvej The North Sea Centre DK-9850 Hirtshals Denmark Tel: + 45 98 944300 Fax: + 45 98 942226 Email: firstname.lastname@example.org/s Norwegian Institute of Fisheries and Aquaculture Ltd Muninbakken 9 . Dublin Eire Ireland Tel: + 353 1 2144100 Web site: www.matis.gov. Baelskaai 25 8400 Oostende Belgium Tel: + 32 59321876 Fax: + 32 59322840 Email: email@example.com/templates/homepage.no/ Matís ohf (Icelandic Fisheries Laboratories) Skúlagata 4 101 Reykjavík Iceland Tel: + 354 422 5000 Fax: + 354 422 5001 Email: firstname.lastname@example.org 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.org/fishery/ European Association of Fish Producers’ Organisations H.com Web site: www.gov.
ifr.bsi-global.ifst.uk Institute of Food Research (IFR) Norwich Research Park Colney lane Norwich NR4 7UA United Kingdom Tel: + 44 (0) 160 3255000 Fax: + 44 (0) 160 3507723 Web site: www.ac.fdf.org.seafood.uk Department of Health Richmond House 79 Whitehall London SW1A 2NS United Kingdom Tel: + 44 (0) 207 2104850 Email: email@example.com/en/index.cmar.org CSIRO Marine Laboratories Castray Esplanade GPO Box 1538 Hobart Tasmania 7001 Australia Tel: + 61 3 62325222 Fax: + 61 3 62325000 Email: firstname.lastname@example.org Other Sources of Information British Standards Institution 389 Chiswick High Road London W4 4AL United Kingdom Tel: + 44 (0) 208 9969001 Fax: + 44 (0) 208 9967001 Email: email@example.com Web site: www.com/ 243 .gov.au Web site: www.org Web site: www.htm Food and Drink Federation (FDF) 6 Catherine Street London WC2B SJJ United Kingdom TeI: + 44 (0) 207 8362460 Fax: + 44 (0) 207 8360580 Email: firstname.lastname@example.org 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 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.CONTACTS Fishing Industry Research Institute 15 Lower Hope Street Rosebank 7700 Republic of South Africa National Fisheries Institute (USA) 7918 Jones Branch Drive Suite 700 McLean VA 22102 United States of America Tel: + 1 703 7528880 Email: email@example.com Web site: www.nz/sc-home Health Protection Agency (HPA) 61 Colindale Avenue London NW9 5EQ United Kingdom TeI: + 44 (0) 208 2004400 Fax: + 44 (0) 208 2007868 Web site: www.uk Web site: www.gsi.
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.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.sfam.co.nhs.scot.00187 Rome Italy Tel: + 39 06 487751 Fax: + 39 06 4877599 Email: email@example.com Web site: www. jsp Food Safety 10 .int/en/ WHO Regional Office for Europe Scherfigsvej 8 DK-2100 Copenhagen Ø Denmark Tel: + 45 39 171717 Fax: + 45 39 171818 Email: firstname.lastname@example.org Codex Alimentarius Commission (CAC) Viale delle Terme di Caracalla 00153 Rome Italy Tel: + 39 (06) 57051 Fax: + 39 (06) 57054593 Email: Codex@fao.uk/index.who.who.int Web site: www.sofht.de 244 .org Web site: www.uk/ Health Protection Scotland Clifton House Clifton Place Glasgow G3 7LN United Kingdom Tel: + 44 (0)141 3001100 Fax: + 44 (0)141 3001170 Email: email@example.com Web site: www.php Society of Food Hygiene and Technology The Granary Middleton House Farm Tamworth Road Middleton Staffs B78 2BD United Kingdom Tel: + 44 (0) 1827 872500 Fax: + 44 (0) 1827 875800 Email: firstname.lastname@example.org Geneva 27 Switzerland Tel: + 41 22 7912111 Fax: + 41 22 7913111 Email: email@example.com/foodsafety World Health Organization (WHO) Headquarters CH .int/ SIK .euro.net/web/indexen. Web site: www.uk/scieh.The Swedish Institute for Food and Biotechnology Box 5401 SE-402 29 Göteborg Sweden Tel: + 46 31 3355600 Fax: + 46 31 833782 Email: firstname.lastname@example.org Web site: www.scot.int Web site: http://www.int.
fpa-food.org/ The Association of Food.CONTACTS US FDA .org/cms/ Useful Web Sites Gateway to Government food safety information (US) http://www.FoodSafety.uk/ Society of Food Hygiene and Technology (UK) http://www.org/ Institute of Food Technologists http://www. Van Buren Suite 1000 Chicago IL 60607 United States of America Tel: + 1 3127828424 or + 1 8004383663 Fax: + 1 3127828348 Email: email@example.com/ Food Science Australia http://www.gov/foodsafety/ Food Safety Authority of Ireland http://www.fsai.fda.gov/ Institute of Food Technologists 525 W.Center for Food Safety and Applied Nutrition 5100 Paint Branch Parkway College Park MD 20740-3835 United States of America Tel: + 1 888 SAFEFOOD (+ 1 888 723 3366) Web site: www.org.csiro.who.riph.sofht.int/foodsafety/en/ European Commission: Activities of the European Union food safety http://europa.co.foodscience.gov/ World Health Organization (WHO): Food safety programmes and projects http://www.au/ International Association for Food Protection http://www.org/ Royal Institute of Public Health (UK) http://www.cfsan. Beverage and Consumer Products Companies http://www.htm Centre for Disease Control and Prevention (CDC) (US) http://www.cdc.ift.eu/pol/food/index_en.org Web site: http://www.uk/ 245 .
FISH AND SEAFOOD 246 .
INDEX Achromobacter spp. breakdown of – as indicator of fish freshness 10-1 Azaspiracids 158-9 detection methods 160 origins and distribution 159 toxicology 159-60 Bacalao 97. as 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. 111 Astroviruses. in crustacean shellfish 87 Alteromonas spp. as autochthonous bacterial flora of fish 3 in fish sauce 127 Bacillus cereus. in crustacean shellfish 86-7 ATP. preserved by dry/wet salting 99 Anisakis 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. 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 . 102 Bacillus spp. growth and survival in crustacean shellfish 86 Air drying 101 Alcaligenes. planktonic source of spirolides in shellfish 163 Allergens. 69-70 Aeromonas hydrophila. growth and survival in chilled and frozen raw fish 19 of concern in preserved fish 115 Aspergillus spp. as spoilage organism in oysters 65 as spoilage organism in tropical crustacean shellfish 81 in molluscan shellfish 60 Alexandrium. to test microbiological criteria 19 Aeromonadaceae. as TTX producers 145 Amnesic shellfish poisoning (ASP) toxins 146-8 caused by domoic acid in crustacean shellfish 87 Anchovies. causing spoilage of dried fish 110. growth and survival in crustacean shellfish 86 Aeromonas sobria. as autochthonous bacterial flora of fish 3 Aeromonas spp. source of PSP toxins 142 Alexandrium ostenfeldii.
fish sauce 123 Basipetospora halophila. of recipe fish meals 43 Cholera. 131 Carbon dioxide. preserved by marinating 100 Chemical detection methods. in crustacean shellfish 86-7 Campylobacter coli. associated with consumption of fish/fish products 16. concern with respect to use of MAP 7 growth and survival in chilled and frozen prepared fish products 45 growth and survival in chilled and frozen raw fish 15-6 growth and survival in crustacean shellfish 84 growth and survival in cured. indigenous – in fish (Table) 4 non-indigenous – in fish (Table) 4 Bakasang. description and uses 56-7 effects of processing on microflora 60-4 hot-smoking 107 Clear smoke treatments 108 Clostridium. growth and survival in crustacean shellfish 86 growth and survival in molluscan shellfish 59-60. 136. for PSP toxins 144 Chemical indicators. as autochthonous bacterial flora of fish 3 Clostridium botulinum. 68-9 Capture and handling. mould causing spoilage of dried fish 110 Battering process 36-8 Bivalves. 113.FISH AND SEAFOOD components of seafood ready meals 40 Bacterial pathogens. as gas in modifiedatmosphere packaging 6 CCP Decision Tree (in HACCP plan) (Fig) 185 Ceviche. 137. importance in lactic acid fermentation in fish products 130. affected by azaspiracids 159 affected by cyclic imines 161 Botulism. caused by Vibrio cholerae O1 17 Ciguatoxins 155-8 detection methods 157-8 origins and distribution 1567 toxicology 157 Clams. of fish – effects on microflora 5-6 Carbohydrate. 226 Breading process 36-8 Brevetoxins. growth and survival in crustacean shellfish 86 pathogenic microorganisms in molluscan shellfish 59-60 Campylobacter jejuni. of fresh fish spoilage 11 Chemical pollutants. 98-9 Brochothrix thermosphacta. smoked and dried fish 113-6 hazard in mild-processed marinated products 101 hazard in sun-dried products 102 non-proteolytic – risk of growth in fish recipe meals 44 248 . in crustacean shellfish 87 Chilled prepared fish products 27-51 Chilled raw fish 1-25 Chilled storage. as cause of spoilage of MAP fish 15 Calciviruses. cause of neurotoxin shellfish poisoning 153-5 origins and distribution 1545 toxicology 155 importance in wet salting of fish 96 Brining 93.
as contaminants in scampi 36. of fish – different processes 94 Dun. to determine fresh fish spoilage 11 Eels. processing 334 Cockles. 149-50. smoked and dried fish 114 Enzymic spoilage. 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. description and uses 59 effects of processing on microflora 60-4 Cold-smoking 104-5. growth and survival in crabmeat 84 growth and survival in cured. as autochthonous bacterial flora of fish 3 Crabs. of prepared fish products 44 Escherichia coli. 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. 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. 106 Contacts 241-5 Conway method. to determine fresh fish spoilage 11 Corynebacteria. as spoilage organisms in tropical crustacean shellfish 81 Coryneforms. adverse condition on poorly preserved fish 110 EC grading scheme.INDEX pathogen profile 226-7 Clostridium perfringens. to detect NSP toxins 155 ELISA kit (Biosense). to detect DSP toxins 150-1 Enrobing process 36-8 Enterobacteriaceae. growth and survival in chilled and frozen raw fish 19 of concern in cold-smoked salmon 115 Domoic acid (DA). for detection of DA 148 ELISA kit (DSP-Check). hot-smoking 107 ELISA assay. of fish – analysis of 10 Diarrhetic shellfish poisoning (DSP) toxins 148-51 detection methods 150-1 origins and distribution 14950 toxicology 150 Dinophysis spp. 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. cooked – effects of processing on microflora 83 forms for purchase 80 freshness indicators 83 Crayfish. source of azaspiracids 159 source of DSP toxins 148-9.
non-B) viruses. for evaluation of fish quality 9 Eurotium. of fresh fish spoilage 911 Evisceration. as autochthonous bacterial flora of fish 3 in molluscan shellfish 60 spoilage organism in oysters 65 Food hygiene legislation 189-224 Food of animal origin. concerning crustacean shellfish 87-8 European Grading Scheme. 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 .FISH AND SEAFOOD EU legislation. as source of inulin in lactic acid fermentation in fish products 134 Garos. in crustacean shellfish 86-7 in molluscan shellfish 59-60 Herring. causing pink condition on fish surface 110 in fish sauce 127 Halococcus spp. specific EU hygiene rules 196-224 Fresh fish. preserved by marinating 101 Gravlaks. to measure fish skin resistance 10 Flavobacterium. in crustacean shellfish 86-7 in molluscan shellfish 59-60 Hepatitis A virus. spoilage 9-11 Freshness grading schemes 9 Frozen prepared fish products 27-51 Frozen raw fish 1-25 Frozen storage. of chilled and frozen prepared fish products – preparation 33-6 Fish sauces 123-4 Fish sauces and pastes. member of cyclic imine group 161 Gymnodinium catenatum. of fish at sea 5 Fatty fish. definition 1 Fermented fish 123-40 Fermented fish/salt products of Thailand (Table) 125 Fish and shellfish toxins 141-74 Fish cakes. as source of ciguatoxins 156 Garlic. of fish – effects on microflora 5-6 Hepatitis (non-A. fish sauce 123 HACCP. 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. source of PSP toxins 142 Gyoshoyu. 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. 102 Gymnodimines. preserved by marinating 100 preserved by dry/wet salting 99 Histamine fish poisoning. fermented fish product 124 General Pathogen Surveillance Programme 84 Gravad fish. 130-5 Fish-spoilage bacteria 12 Fishtester. processing 35 Fish component. fermented fish product 126 Green cured split cod 97. of prepared fish products 44 Gambiertoxins.
131 Lactic fermented fish products (Table) 126 fish species used in the Philippines (Table) 126 Lactobacillus. cooked – effects of processing on microflora 83 forms for purchase 80 freshness indicators 83 Mackerel. leading to oxidative rancidity 11 Lipophilic shellfish poisoning toxins 149. 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 source of yessotoxins 152 Lipid content.INDEX Hygiene regulations for foodstuffs production and distribution (EU) 190-6 Ice storage. initial – of chilled and frozen prepared fish products 31-2 of chilled and frozen raw fish 3-5 251 . hot-smoking 107 preserved by dry/wet salting 99 Marinating 94. 100-1 Masmeen 107 Matjes. concerning crustacean shellfis 88 Jellied eels. as autochthonous bacterial flora of fish 3 Lean fish. causing spoilage in vacuum-packed dried salmon 111 Lactic acid fermentation. 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 Karenia brevis. for chilled and frozen prepared fish products 47-9 for chilled and frozen raw fish 19-20 for crustacean shellfish 87-8 for cured. smoked and dried fish 114-5 hazard in cold-smoking 105 hazard in mild-processed marinated products 101 Listeriosis. as source of botulism 136 International legislation. growth and survival in chilled and frozen prepared fish products 46 growth and survival in cured. preserved by marinating 101 Microbiological criteria. outbreak caused by gravid 115 Lobster. definition 1 Legislation. preserved by salt-boiling 100 Ionising radiation. for fresh and frozen fish (Table) 20 Micrococcus spp. 151-3 detection methods 153 Liquid smoke 107-8 Listeria spp. food hygiene 189-224 Lingulodinium polyedrum. in fish fillet 1 Lipid oxidation. enabled by carbohydrate source in fish products 130. smoked and dried fish (Table) 116-7 for shellfish products 70 Microbiological limits. 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. 135 I-zushi.
as TTX producer 145 Norovirus. reduction in smoking process 108 Norcardiopsis dassonvillei. as autochthonous bacterial flora of fish 3 as spoilage organism in temperate crustacean shellfish 80 in molluscan shellfish 60 Mouse bioassay. contaminated by azaspiracids 158 description and uses 58 effects of processing on microflora 60-4 hot-smoking 107 Nam-budu. fish sauce 123 Neosaxitoxin (neoSTS) 142-3 Neurotoxin shellfish poisoning (NSP) toxins 153-5 detection methods 155 Nitrosamines. contamination leading to illness 68 description and uses 57 effects of processing on microflora 60-4 hot-smoking 107 spoilage organisms 65 P. smoked and dried fish 95 of fermented fish 126-7 of molluscan shellfish 59-60 Microflora. caused by lipid oxidation 11 Oysters. growth and survival in chilled and frozen raw fish 17 Paralytic shellfish poisoning (PSP) toxins 142-4 detection methods 144 toxicology 143 252 . in crustacean shellfish 867 Norwalk virus. pathogenic microorganisms in molluscan shellfish 59-60 Opisthorchis viverrini. associated with consumption of fermented fish products 137 Oxidative rancidity. 153 for detection of LST toxins 151 for detection of NSP toxins 155 for detection of pinnatoxins and pteriatoxins 165 for detection of PSP toxins 144 for detection of spirolides 164 for detection of tetrotoxin 145 Mussels.FISH AND SEAFOOD of crustacean shellfish 80-1 of cured. 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. shigelloides. for detection of azaspiracids 160 for detection of ciguatoxins 157 for detection of DA 148 for detection of DSP toxins 150. of crustacean shellfish – effects of processing 81-3 of cured.
as source of yessotoxins 152 Protoperidinium cassipes. as component for seafood ready meals 40 Prawns. effects of processing on microflora 81-2 freshness indicators 83 Pre-column derivatisation. causing spoilage in vacuum-packed dried fish 111 Pseudomonads. 225-39 Pathogens. as cause of histamine fish poisoning 15 as spoilage organism in chilled products 43 as spoilage organism in fresh fish 14-5 Pickling 94. mould causing spoilage of dried fish 110 Potato. as source of toxins 159. 100-1 Pindang. spoilage organism in oysters 65 Protoceratium reticulatum. preserved by salt-boiling 100 Pink. growth and survival in chilled and frozen raw fish 19 growth and survival in crustacean shellfish 86 growth and survival in cured. in fish fillet 1 Proteus. 152-3 Penicillium spp. 69-70 Polycyclic aromatic hydrocarbons. pathogen profile 230 in molluscan shellfish 5960. 145-6 definitions 28-30 non-seafood ingredients 32 Processing. contact details 241-2 Prorocentrum spp. 108 Polypaecilum pisce. 163. 197 Plesiomonas. in molluscan shellfish 60 Pseudomonas spp. as component for seafood ready meals 40 Pathogen profiles. reduction in smoking process 103. planktonic source of azaspiracids 159 Pseudomonadaceae. 161. effects on microflora of fish 5-9 Professional bodies. detection methods 165 origins and distribution 165 toxicology 165 Pissala. fermented fish product 124 Plankton. as autochthonous bacterial flora of fish 3 as spoilage organism in chilled products 43 as spoilage organism in crustacean shellfish 80. causing spoilage of dried fish 110-1 Photobacterium phosphoreum. 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. as source of DSP toxins 148-9 Protein content.INDEX Parasites. adverse condition on poorly preserved fish 110 Pinnatoxins member of cyclic imine group 164. smoked and dried fish 115 Pasta. for detection of PSP toxins 144. smoked and dried fish 113-6 growth and survival in fermented fish 136-7 growth and survival in molluscan shellfish 66-70 Pectenotoxins 151. 81 as spoilage organism in fresh fish 13 as spoilage organism in oysters 65 as TTX producers 145 253 .
as source of tetrotoxin 144-5 Putty fish.FISH AND SEAFOOD inhibition of growth by carbon dioxide 6 Pseudonitzschia multiseries. detected in cod 115 Pteriatoxins. as carbohydrate source in lactic acid fermentation in fish products 134 as component for seafood ready meals 40 RIDASCREEN. as spoilage organism in chilled products 43 as spoilage organism in fresh fish 13-4 as spoilage organism in temperate crustacean shellfish 80. smoking and marinating of fish 96 Salt-boiling 94. chilled and frozen 1-25 Recipe dish. detection methods 165 member of cyclic imine group 164 origins and distribution 165 toxicology 165 Puffer fish. for evaluation of fish quality 9 Radiation. processing and effects on microflora 60-1 Shewanella. as hazard in regard to reef-caught fish 28 growth and survival in crustacean shellfish 84 growth and survival in cured. 100 Salting 97-9 definitions 93-4 Sardines.) 41 ingredients preparation. for fish ready meals – manufacture 40-2 Saxitoxin (STX) 142-3 Scallops. assembly and packaging. preserved in saturated brine 99 Sauce. implicated in amnesic shellfish poisoning 146-7 Pseudoterranova. initial microflora 31 Seafood ready meals 39-40 Seafood recipe meal centres 38-9 Shaped fish products. 36 Seafood. in molluscan shellfish 60 Rice. description and uses 58-9 Scampi. source of PSP toxins 142 Quality Index Method. fermented fish product 126 as source of botulism 136 Rakørret. resulting from poorly salted cod 110 Pyrodinium. 135 Rakefisk.) 41 Recipe fish products 38-40 Rhodotorula rubra. to measure fish skin resistance 10 Salmon. as autochthonous bacterial flora of fish 3 inhibition of growth by carbon dioxide 6 Shewanella putrefaciens. processing 34. for microbial inactivation in fish products 127. growth and survival in chilled and frozen raw fish 19 Pseudoterranova dicipiens. importance in lactic acid fermentation in fish products 130 importance in salting. vacuum packing 105 Salmonella spp. producers of brevetoxins 154 Raw fish. antibody-based kit to detect STX 144 RT Freshmeter. fermented fish product 126 Raphidophytes. flow diagram (Fig. 83 254 . processing 35 Shellstock. smoked and dried fish 114 pathogen profile 230-1 pathogenic microorganisms in molluscan shellfish 59-60 Salt burn. hazard in salting of fish 97 Salt concentration. flow diagram (Fig.
in poorly salted cod 110 Smoked fish 93-121 Smoking 103-8 different processes 94-5 Snow Mountain virus. fish sauce 123 Shrimps. of fish after capture 5 Surstrømming. in molluscan shellfish 60 Tuna. fish and shellfish 141-74 Trade associations. of dried fish 111 of fish 6-7 of fish – (US) NACMCF recommendations 7-8 of fish – Codex Committee 255 . 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. 14 Torry Scale. fermented fish product 126 Taste panel evaluation.INDEX Shigella spp. of cooked fish 9-10 Tetrotoxin 144-6 detection methods 145-6 toxicology 145 TMAO. concerning crustacean shellfish 89 Vacuum packaging. of crustacean shellfish 80. growth and survival in chilled and frozen prepared fish products 46 growth and survival in cured. as measure of fish degradation 10 Sliming. to measure fish skin resistance 10 Total Volatile Base-Nitrogen. for evaluation of fish quality 9 Torrymeter. detection methods 164 member of cyclic imine group 162-4 origins and distribution 163 toxicology 163-5 Spoilage. grading of 31 hot-smoking 107 US legislation. 12-5 of recipe fish meals 43-4 of shellfish 65 Staphylococcus. as indicator of fresh fish spoilage 11 Toxins. 13. contact details 241-2 Trematode infections foodborne 137 Trichosporon. processing and effects on microflora 63-4 Skin resistance. role in fish degradation 10. definitions of forms for purchase 79-80 effects of processing on microflora 81-2 freshness indicators 83 Shucked shellfish. 101 Storage. pathogenic microorganisms in molluscan shellfish 59-60 Specific spoilage organisms (SSOs) 12-5 Spirolides. as contaminant in scampi 36 as contaminant in enrobing process 37 as hazard in dry salting of fish 89 growth and survival in chilled and frozen prepared fish products 46-7 growth and survival in crustacean shellfish 84 growth and survival in cured. 83 of fermented fish 135-6 of fish subjected to different preservation methods 108-10 of fresh fish 9-11. smoked and dried fish 114 pathogenic microorganisms in molluscan shellfish 59-60 Shottsuru.
processing for seafood recipe meal centres 39 Vibrio spp. growth and survival in chilled and frozen prepared fish products 47 growth and survival in crustacean shellfish 86-7 Wallemia sebi. of shellstock 61. growth and survival in chilled and frozen raw fish 16-7 growth and survival in crustacean shellfish 86 growth and survival in fermented fish 136 growth and survival in molluscan shellfish 66-67 pathogen profile 234-5 Vibrio vulnificus. as hazard in regard to reef-caught fish 28 as spoilage organism in crustacean shellfish 80. 68 Vibrio cholerae non-01. as spoilage organisms in chilled recipe meals 43 Yessotoxins 151. 152-3 256 .FISH AND SEAFOOD guidance 7 of fresh chilled products 42 Vegetables. as autochthonous bacterial flora of fish 3 as spoilage organisms in chilled products 43 as spoilage organisms in vacuum-packed dried fish 111 Viruses. growth and survival in chilled and frozen raw fish 17 Vibrio cholerae 01 and 0139. growth and survival in molluscan shellfish 67-8 Vibrio parahaemolyticus. causing dun condition on dried fish 110 Wet salting 98-9 Wet storage. 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. 62 Yersinia spp. 81 as TTX producers 145 pathogenic microorganisms in molluscan shellfish 59-60 risk in cold-smoking 105 Vibrio cholerae. growth and survival in crustacean shellfish 85 pathogen profile 233-4 Vibrio cholerae 01.