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Leatherhead Food International Leatherhead Food International
MICROBIOLOGY HANDBOOK FISH AND SEAFOOD
This edition first published 2009 by Leatherhead Publishing, a division of Leatherhead Food International Ltd Randalls Road, Leatherhead, Surrey KT22 7RY, UK URL: http://www.leatherheadfood.com and Royal Society of Chemistry Thomas Graham House, Science Park, Milton Road, Cambridge, CB4 0WF, UK URL: http://www.rsc.org Regstered Charity No. 207890
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Associate Prof. Covadonga Arias Microbial Genomics Department of Fisheries and Allied Aquacultures Auburn University Auburn Alabama 36849 United States of America Dr. Simon Derrick Special Projects The Grimsby Institute Humber Seafood Institute Europarc Grimsby DN37 9TZ United Kingdom Carlos Abeyta, Jr. Supervisory Microbiologist Pacific Regional Laboratory Northwest U.S. Food and Drug Administration 22201 23rd Dr. S.E. P. O. Box 3012 Bothell WA 98021-4421 United States of America Linda Nicolaides, M.Ph., FRSPH Ethical Trade and Food Management Group Natural Resources Institute University of Greenwich Central Avenue Chatham Maritime Kent ME4 4TB United Kingdom
Prof. Martin Adams Faculty of Health and Medical Sciences University of Surrey Guildford Surrey GU2 7XH United Kingdom Jeffrey L.C. Wright C.M., Ph.D., FCIC Carl B. Brown Distinguished Professor of Marine Science UNCW Center for Marine Science Marvin Moss Lane Wilmington NC 28409 United States of America Rhea Fernandes and Dr. Peter Wareing Leatherhead Food International Randalls Road Leatherhead Surrey KT22 7RY United Kingdom Eugenia Choi and Dr. Jenny Pfleger Regulatory Advisors Leatherhead Food International Randall Road Leatherhead Surrey KT22 7RY United Kingdom
and Meat Products. and Ann Pernet for the work put into typesetting and indexing. and growth characteristics for key pathogens associated with the product. I thank the contributing authors for their collaboration. guides to HACCP. Acknowledgements All the authors involved in writing the first edition of this book are gratefully acknowledged.Goldwyn. Rhea Fernandes Leatherhead Food International v . both in the UK and elsewhere. The books in the series are designed as easy-to-use guides to the microorganisms found in foods.FOREWORD The Microbiology Handbook series includes Dairy Products. and a detailed list of contacts for various food authorities. published by Leatherhead Food International and RSC Publishing. for his scientific input and Victoria Emerton. Each book provides a brief overview of the processing factors that determine the nature and extent of microbial growth and survival in the product. which has been the chief factor in ensuring the completion of this book. All handbooks also contain a review of the related legislation in Europe and UK. I dedicate this book to my husband . Fish and Seafood. My gratitude is extended to Dr Peter Wareing. Training Manager. The books are intended as a source of information for microbiologists and food scientists working in the food industry and responsible for food safety. thank you for all the support. potential hazards associated with the consumption of a range of products. I also acknowledge and thank Jackie Apps. Technical Team Leader for her valuable editorial contribution. Alison Turner.
2 Initial Microflora 4.5 Factors Affecting Fresh Fish Spoilage 1.7 References 3. 3.CONTENTS CONTRIBUTORS FOREWORD INTRODUCTION 1.4 Spoilage 3.3 Processing and its Effects on the Microflora 4.7 Published Microbiological Criteria 2.4 Spoilage 1.4 Spoilage 4.8 References 1. 4.1 Introduction 2.3 Initial Microflora 2.6 Published Microbiological Criteria 4. CHILLED AND FROZEN RAW FISH 1.7 References 4.5 Spoilage 2.1 Definitions 1. vii .8 Further Reading CRUSTACEAN SHELLFISH 4.8 Further Reading iii v xi 1 1 3 5 9 12 15 19 20 24 27 27 28 31 32 43 45 47 49 53 53 59 60 65 66 70 71 77 79 79 80 81 83 84 87 89 92 2.5 Pathogens: Growth and Survival 3.9 Further Reading CHILLED AND FROZEN PREPARED FISH PRODUCTS 2.1 Definitions 3.5 Pathogens: Growth and Survival 4.7 Published Microbiological Criteria 1.3 Processing and its Effects on the Microflora 1.2 Initial Microflora 3.6 Pathogens: Growth and Survival 1.6 Published Microbiological Criteria 3.2 Definitions 2.4 Processing and its Effects on the Microflora 2.6 Pathogens: Growth and Survival 2.2 Initial Microflora 1.3 Processing and its Effects on the Microflora 3.8 References MOLLUSCAN SHELLFISH 3.1 Definitions 4.
SMOKED AND DRIED FISH 5.5 Regulation (EC) No.1 Definitions 5. S.9 Azaspiracids 7.11 Conclusion 7. 8.3 Regulation (EC) No.9 Other Relevant Legislation 223 9. 2006 No. 2073/2005 on Microbiological Criteria for Foodstuffs 214 9.6 Regulation (EC) No.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. 852/2004 on the General Hygiene of Foodstuffs 191 9. 14 (Hygiene requirements specific to the UK) 221 9.1 Introduction 7.4 Amnesic Shellfish Poisoning (ASP) Toxins 7.2 Initial Microflora 6.10 Cyclic Imines 7.7 Food Hygiene (England) Regulations 2006.4 Implementation and Review of the HACCP Plan 8.6 References 6. 853/2004 Laying Down Specific Hygiene Rules for Food of Animal Origin 196 9.10 Further Reading 223 viii .2 Paralytic Shellfish Poisoning (PSP) Toxins 7.2 Definitions 8.2 Initial Microflora 5.4 Spoilage 5.6 Published Microbiological Criteria 5. 9.7 Neurotoxin Shellfish Poisoning (NSP) Toxins 7.4 Spoilage 6.5 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 6.3 Processing and its Effects on the Microflora 6.1 Introduction 189 9. EU FOOD HYGIENE LEGISLATION 189 9.5 Pathogens: Growth and Survival 5.8 Ciguatoxins 7.1 Definitions 6.3 Stages of a HACCP Study 8.I.12 References HACCP IN FISH AND SEAFOOD PRODUCT MANUFACTURE 8. 7.5 Pathogens: Growth and Survival 6.2 Legislative Structure 190 9.1 Introduction 8.7 Further Reading FISH AND SHELLFISH TOXINS 7.3 Processing and its Effects on the Microflora 5.7 References 5. CURED.5 Diarrhetic Shellfish Poisoning (DSP) Toxins 7.3 Tetrodotoxin 7.8 Further Reading FERMENTED FISH 6.8 Guidance 222 9.6 Lipophilic Shellfish Toxins (LST) 7.4 Regulation (EC) No.
10.8 Vibrio cholerae 10.11 References 225 225 226 227 229 230 230 232 233 234 235 236 241 247 CONTACTS INDEX ix .1 Aeromonas spp.6 Salmonella spp.10 Vibrio vulnificus 10.10 PATHOGEN PROFILES 10.4 Listeria spp.2 Clostridium botulinum 10.3 Clostridium perfringens 10.7 Staphylococcus aureus 10.5 Plesiomonas 10. 10.9 Vibrio parahaemolyticus 10. 10.
Milk.A. Seafood Research from Fish to Dish: Quality. Clark S. Hui Y. for fish. and fish and shellfish toxins. Further Reading General Fish/Seafood Microbiology Cross N. Eds. Seafood and Vegetables.H. the consumption of fish and seafood has increased. Oehlenschlager J. crustacean shellfish. Cross N. Wageningen. Seafood processing: basic sanitation practices.. xi .INTRODUCTION Fish and seafood are a main source of animal protein in the diet. Wiley. Poultry. and. 2007. whether they are pelagic (surface to mid-water) or demersal (bottom) feeders will influence the numbers and types of microorganisms present on freshly caught seafood. cured. chilled and frozen prepared fish. molluscan shellfish..B. season. Jacobsen C. 2006. Chandan R. The Microbiology Handbook. Nollet L. Key changes in this edition are the recent regulatory changes pertaining to food hygiene and microbiological criteria for foodstuffs. smoked and dried fish. Wageningen Academic Publishers. Meat. fish and seafood can be involved in the transmission of pathogenic microorganisms and toxins. Safety and Processing of Wild and Farmed Fish. Geographical region.. The second edition of this handbook is a review of the entire book for currency of information.Fish and Seafood consists of the microbiology of seven different product categories: chilled and frozen raw fish. Saebo A. 94767. thus..M.. Hoboken.. Luten J.. in Handbook of Food Products Manufacturing: Health. fermented fish.. Catches can be gathered from seas. rivers and lakes whose water can range from pristine to contaminated. Bekaert K. Often contamination is from human and animal sources. Because of their health advantages over red meats.
431-43. 1997.. 372-6. Woodhead Publishing Ltd. Woodhead Publishing Ltd. in Advances in Food and Nutrition Research.. Noordwijkerhout. Gudmundsson M. 60 (4). 87121. Stanfield P.H.. Luten J. 2005. basic sanitation practices. Eds. Ed. The incidence of Listeria spp. Hui Y.R.N. in Food Plant Sanitation.H. Huss H. New York. Kluwer Academic / Plenum Publishers. 121-37. 543-61. Minimal processing in practice: seafood. London. Amsterdam. Ed. Eds. Biogenic amines in fish and shellfish. 1998. November 1995. 61 (3). 2004. 1996. Salmonella spp.. 36 (1-2). Taylor S. London. in Kosher Food Production. 459-72.H. McCarthy S. 1996.A. Rawles D. Ashie I. Bruinsma B. Academic Press.. 413-30. 318-23. Journal of Food Protection. 245-66 Bremner H. Smith J.L. Proceedings of the International Seafood Conference. Flick G. 8 (2).. Food Control. Simpson B. Davies A. Ventresca P. 2004. Smith J... Tong P. Hall G.B. Safety and Quality Issues in Fish Processing. Cambridge. Ed. in Seafood from Producer to Consumer: Integrated Approach to Quality.M. Predictive microbiological modelling and seafood quality. Ed. 329-65.Y. 1996.E. Ed.. Blackwell Publishing. Blech Z. Gram L.. Ed. CRC Critical Reviews in Food Science and Nutrition. Eds. Hafsteinsson H.K. Hui Y. Noordwijkerhout.J. Spoilage and shelf-life extension of fresh fish and shellfish. Stanfield P.B. Incidence and survival of Listeria monocytogenes in ready-to-eat seafood products. Huss H. Modified-atmosphere packaging of fish and fish products.S.. Control of indigenous pathogenic bacteria in seafood.A. Seafood: processing.. Martin R. 2003. in Seafood from Producer to Consumer: Integrated Approach to Quality.. Seafood processing: basic sanitation practices. 200-23.R.L. in Food Processing: Principles and Applications. and Clostridium botulinum in smoked fish and shellfish. Johnson J..International Commission on Microbiological Specifications for Foods. 2002. in Fish Processing Technology..S. Nip W-K. 33 (1). Blech Z.D. Dalgaard P. Dalgaard P.P. Gram L.Y. Oxford. Luten J. in Minimal Processing Technologies in the Food Industry.L. 91-8. November 1995. New York. Proceedings of the International Seafood Conference. Blackwell Publishing. in Microorganisms in Foods 6: Microbial Ecology of Food Commodities. International Commission on Microbiological Specifications for Foods. Microbiological spoilage of fish and fish products. Huss H. Bengtsson N. Microbiology of fish and fish products. Vo1ume 39. 1997. Amsterdam.H. 1997.. Journal of Food Protection. 174-249. The fish industry. Oxford.A.. 167-77. 2002. Ohlsson T. International Journal of Food Microbiology. Elsevier. 1997. 1997.H. Cambridge.. xii . Blackie. Elsevier. Marcel Dekker.M. Heinitz M. Fish and fish products (Microorganisms in fish products). Gorham J.
Ed.S. 166-73. Gram L. Huss H. International Journal of Food Microbiology. August 1991. Jakobsen M. 26 (3). Sterling Publications Ltd. 265-79. Huss H. Evaluation of the bacteriological quality of seafood. 1995. 319-33. Jakobsen M. Blackie. Food Science and Technology Today. Copenhagen. Elsevier. Eds.H. Dutson T. Microbial hazards of seafood consumption. Dalgaard P. PHLS Microbiology Digest. 2257. London.N. 1990. Bonnell A. 1991. xiii . in Food and Beverage Technology International: USA. Institut fur Veterinarmedizin. Ed. poultry and fish. in Quality Assurance in Seafood Processing: A Practical Guide. 359-403. Volume 5: Microbiological Specifications of Food Pathogens.. guidelines. 1994. Liston J.D. Olley J. Qualitative and quantitative characterization of spoilage bacteria from packed fish. Sofos J. Jakobsen M. Ross T..D. 459-78. Bacterial spoilage of seafood. Liston J. Microbial growth and its control in meat. Gram L.. Garrett E. 46 (1). 16 (1). 93-105. 1994. Microbiological quality of fishery products. London. 5662. in Quality Assurance in the Fish Industry: Proceedings of an International Conference. London. Copenhagen. Eds. Application of predictive microbiology to assure the quality and safety of fish and fish products. 44 (12).M. 5 (3). Amsterdam.A. 1992.M.H. Twigg B. International Journal of Food Microbiology. Amsterdam. 118-20. Ed. 11 (2). 90-106.M. in Quality Attributes and Their Measurement in Meat.. The microbiology of seafood products. Microbiological indicators and seafood processing. A new look at microbiological standards. 1992. 26 (3). 1995. Amsterdam. London.J. Ward D. Seafood toxins of microbiological origin. Elsevier.A. Ward D. in Quality Assurance in the Fish Industry: Proceedings of an International Conference. Liston J. 269-82. Pearson A.International Commission on Microbiological Specifications for Foods. 1-17. Bonnell A. Hobbs G. Ed. 1994..R. 1989. in Public Health Aspects of Seafood-borne Zoonotic Diseases. in Fisheries Processing: Biotechnological Applications... Ed. Poultry and Fish Products. Berlin.R. Fish: microbiological spoilage and safety. Food Technology. Gibson D. Fish spoilage and safety. Liston J.. Copenhagen. 1994.H. in Microorganisms in Foods. Bernoth E-M. Chapman & Hall. Modelling of microbial activity and prediction of shelf-life for packed fish. International Journal of Food Microbiology. Eds. Bacterial pathogens transmitted by seafood. 1990. Eds. and specifications for seafood products. International Commission on Microbiological Specifications for Foods. Dalgaard P. 121-8. Chapman and Hall. Huss H. Martin A. Elsevier. 1992. Activities Report of the R&D Associates. 1996. August 1991. Liston J. Glasgow. McMeekin T. Blackie. 25-39.. Evaluation of the bacteriological quality of seafood. in Quality Assurance in the Fish Industry: Proceedings of an International Conference.R. 1992. 1994. Gilbert R. 54-61. August 1991. 305-17.
1988.K. London.M.R. Robinson R. Volume 1. Food Technology. Elsevier Applied Science Publishers. Simmonds C. in Developments in Food Microbiology. Wiley. guidelines. Part I . 1986. Regenstein J.spoilage of fish. Applied Science Publishers.S. London. Microbiology of frozen fish and related products. in Essays in Agricultural and Food Microbiology. 103. 1988. 104-9. Food Technology. 1982. 117-27. Ed. Norris J.J. 1985. 169-208.. 11 (2)..R. Eyles M. Hackney C. 42 (3). Hobbs G. 1988. and specifications and inspection of seafood products. 71-117. 1987. Microbiological hazards associated with fishery products. 42 (3). in Microbiology of Frozen Foods. xiv . Dicharry A.C. Microbiology of fish.Pedrosa-Menabrito A.K. 199-226.. Microbiological standards. Chichester. Garrett E. 46 (1). CSIRO Food Research Quarterly.a review. 8-16. Hodgkiss W. The bacteriology of fish handling and processing. Journal of Food Quality. Ed. Seafood-borne bacterial pathogens of marine origin. Hobbs G.. 90-3. Davies R. Ed. Shelf-life extension of fresh fish . Lamprecht E.
However. In addition. CHILLED AND FROZEN RAW FISH Associate Prof. which can be up to 67%. Covadonga Arias Department of Fisheries and Allied Aquacultures Auburn University Auburn Alabama 36849 United States of America 1. it shows a typical season pattern especially in migratory species such as herring or mackerel. The lipid content. and spawning season. fins and a streamline body. The connective tissue content of fish is also lower than that found in meat. In addition. while fatty fish have fat cells distributed along their bodies. and excludes molluscs and Crustacea.5% (1). 1 .20%. In this chapter. This super class of vertebrates includes all the bony and cartilaginous finfish. some regulatory agencies such as the US Food and Drug Administration (FDA) will include molluscan shellfish. crustaceans. and is mostly interspersed between the muscle fibres. Muscle composition and structure of fish also differ from those found in other meat. Fish are an important part of a healthy diet since they contain high quality protein. Fish fillet composition can vary significantly within the same species due to feed intake. most fish contain omega 3-fatty acids and other essential nutrients. The lipid fraction is the component showing the greatest variation. offering less than 0. giving it its characteristically flaky structure.1 Definitions Fish are classified as any of the cold-blooded aquatic vertebrates of the super class Pisces typically showing gills. Fish fillets are a poor source of carbohydrates.21%. Protein content in fish fillet varies typically from 16 . Fish can be divided into fatty and lean fish. respectively (1). migratory patterns.1. typically 3 and 15% of total weight. ‘fish’ also refers to the flesh of such animals used as food.2 . Although fish is broadly similar in composition and structure to meat there are a number of distinctive features. and other forms of aquatic animal as part of their ‘fish’ definition. “fish” will be used for fresh and seawater finfish. Fish flesh is dominated by the abundance of white muscle in relatively short segments. lean fish are those fish that store most of their fat in the liver. typically fluctuates between 0. but typically present a low fat percent when compared to other meats.
frozen or preserved. Organoleptic refers to qualities such as appearance. at -2 to -4 °C. Modified-atmosphere packaging (MAP) refers to packaging systems in which the natural gaseous environment around the product is intentionally replaced by other gases. This technique is primarily used for bulk storage. odour and texture. through their development on fish. and texture of the fish. but not below 3 °C during storage. firmness. DMA is dimethyl amine. nitrogen (N2) and oxygen (O2).FISH AND SEAFOOD Chilled fish is fish that has been cooled to. spoilage microflora describes an association of microorganisms that. hence. and maintained at or below -2 °C (normally below -12 °C) during storage. during storage. transportation and sale. Quality refers to palatability and organoleptic characteristics such as tenderness. renders that fish objectionable to consumers. transportation and sale. Shelf life is defined as the time of storage before microbial spoilage of a fish is evident. marbling. and maintained at temperatures just below the freezing point. Fresh fish is raw fish that has not been processed. transportation and sale. Controlled-atmosphere packaging (CAP) refers to packaging in an atmosphere where the composition of the gases is continuously controlled during storage. and flavour based on the maturity. but no further control is exercised during storage. Superchilled fish is fish that has been cooled to. juiciness. and maintained at or below 7 °C. usually carbon dioxide (CO2). Frozen fish is fish that has been cooled to. 2 . Spoilage describes changes that render fish objectionable to consumers. colour. colour. Spoilage potential is a measure of the propensity of microorganisms to render fish objectionable to consumers through the production of offensive metabolic byproducts. The proportion of each component is fixed when the mixture is introduced. Evisceration is the removal of the viscera from a fish. Raw fish refers to fish that has not been cooked but excludes fish treated with curing salts and/or subjected to fermentation.
I). Flavobacterium.II) (3). whenever possible. and those that although not autochthonous to the aquatic environment. However. In fact. It must be noted that mesophilic bacteria can rapidly overgrow psychrotropic organisms. molecular-based methods have not yet been used to address this issue. healthy fish is considered sterile and should not present any bacteria or other microorganisms. Gastrointestinal tracts and gills typically yield high bacteria numbers. For example.2 Initial Microflora The subsurface flesh of live. although these are influenced by water quality and feed. It is crucial to mimic the environmental physicochemical parameters when isolating bacteria from fish. 3 . since they heavily depend on the type of sample analysed and the protocol used for isolation. It is difficult to estimate how many microorganisms are typically associated with fish. The number and diversity of microbes associated with fish depend on the geographical location. Human pathogenic bacteria can be part of the initial microflora of fish. some species (most Vibrios) require sodium chloride for growth. microorganisms colonise the skin. potential human pathogens may be present in both scenarios. More accurate. In general. several culture media containing sodium chloride and more than one incubation temperature should be used. posing a concern for seafoodborne illnesses (2). Members of the families Vibrionaceae (Vibrio and Photobacterium) and the Aeromonadaceae (Aeromonas spp. are present there as result of contamination (anthropomorphic origin or other) or are introduced to the fish during harvest. Fish harvested from clean and cold waters will present lower bacterial numbers than fish from eutrophic and/or warm waters. standard culture-dependent methods can only recover between 1 to 10% of total bacteria present in any given sample. as with other vertebrates. gills and the gastrointestinal tract of fish. Gram-positive organisms such as Bacillus. Shewanella and Pseudomonas. processing or storage (Table 1. Moraxella. On the contrary.CHILLED AND FROZEN RAW FISH Vacuum packaging (VP) refers to packaging systems in which the air is evacuated and the package sealed. the natural fish microflora tends to reflect the microbial communities of the surrounding waters. These pathogens can be divided into two groups: organisms naturally present on fish (Table 1. Lactobacillus and coryneforms can also be found in varying proportions (1). the season and the method of harvest. The autochthonous bacterial flora of fish is dominated by Gram-negative genera including: Acinetobacter. Clostridium. and typical of the fish flora. Micrococcus. 1.) are also common aquatic bacteria.
0.37 °C High for most species.>106 Salmonella spp. stream water. Plesiomonas shigelloides 3 . Escherichia coli Yersinia enterocolitica 101 .109 cells/g) * Some sources considered L. 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.37 °C Unknown Unknown TABLE 1. and mud Ubiquitous. human origin Minimum Infective Dose Variable depending on the strain (>102 cells/g) Staphylococcus aureus 105 . Vibrio cholerae Vibrio parahaemolyticus Vibrio vulnificus other vibrios Aeromonas spp. Toxin levels 0. birds. 4 .19 µg/kg bodyweight from < 102 . and risk associated with the consumption of seafood is low (2. numbers of pathogenic bacteria in raw fish tend to be low.106 cells/g.II Non-indigenous bacterial pathogens frequently present in fish (3) Organism Listeria monocytogenes* Primary habitat Soil. estuarine environments.26 °C 10 . vulnificus) 5 . sewage.103 High (107 . during storage indigenous spoilage bacteria tend to outgrow potential pathogenic bacteria.106 cells/g (exception: V.14 . 105 . 4).I Indigenous bacterial pathogens typically present in fish (3) Organism Temperature range Estimated Minimum Infective Dose 00. Intestinal track of terrestrial vertebrates Human origin Fecal contamination Ubiquitous in environment Shigella spp.35 °C 8 .FISH AND SEAFOOD TABLE 1. However.1 .102 101 . In addition.1 µg toxin lethal dose Clostridium botulinum non-proteolytic type E Pathogenic Vibrio spp.
and spoilage. Of particular concern is the use of wooden or soiled plastic containers for storage and unloading at the quayside.a practice that may have both advantages and disadvantages. This contrasts greatly with the meat industry. The action of intestinal enzymes and activity of the gut bacteria on the flesh around the belly cavity may produce discolouration. and conditions of storage. the fish may be stored in the vessel for periods ranging from just a few hours to several weeks in melting ice. boxes and/or baskets. care should be taken in removing all the gut contents and washing the carcass thoroughly prior to refrigerating.3. in which the bacterial load 5 . Physical damage such as loss of scales. In addition. however. offering better control of fish health prior to. digestion and off-flavours in uneviscerated fish. decks. Used refrigerated brines can be contaminated with high numbers of psychrotrophic spoilage bacteria.3 1.CHILLED AND FROZEN RAW FISH Shelf life depends on the initial microflora on the fish. The length of time that set nets have been in the water or the time trawlers’ nets are towed. with very little control over the condition of the fish at the time of death or the duration of the killing process. Inadequate circulation of chilled brines may result in localised anaerobic growth of some microorganisms. During capture and storage. hook and line. with fish such as tuna and cod being more commonly eviscerated than sardines. 1. and especially when fish is stored on board for longer periods. chilled brine or refrigerated seawater at -2 °C. in recent decades. birds and soil. with the production of off-odours. icing or freezing. bruising and bursting of the gut will increase the number of sites open for bacterial attack and spread. Increasingly. However. and during harvest. ropes. in which the health of each animal can be assessed prior to slaughter. aquaculture practices have been expanding worldwide. has an effect on the amount of stress and physical damage that the fish will suffer during capture. If evisceration is carried out at sea. After capture. potential contaminants added during handling and processing. In eviscerated fish. finfish will almost invariably come into contact with nets. These contacts will not only increase the bacterial cross-contamination between fish batches but will introduce microorganisms from other sources such as humans. or traps. human hands and clothing. handling and processing Wild finfish are usually caught by net. Fish may be eviscerated prior to storage at sea . cortisone levels increase during prolonged stress and can alter the fillet quality. and the killing process is designed to minimise stress.1 Processing and its Effects on the Microflora Capture. freezing facilities (-18 °C) may be used to prevent the catch from deteriorating. the cuttings provide areas of exposed flesh that are open to microbial attack. 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 their re-use will increase the crosscontamination of other fish with such microorganisms. mackerel or herring.
ferric ion (Fe3+)). however. fish have a higher post mortem pH. Bacterial composition under MAP shifts from mostly Gram-negative to predominantly Gram-positive (lactic) bacteria. in time. and minimising handling will all reduce microbial cross-contamination. a reduction in the oxygen tension is achieved which. cleaning of the work environment. These changes can be deleterious. The atmospheric conditions surrounding a product may be passively or actively altered. These containers are also used for displaying the fish during auction at the quayside. 1. Brochothrix thermosphacta and psychrotrophic lactic acid bacteria (LAB) can produce spoilage characteristics. will result in fully anoxic conditions. Both carbon dioxide and low pH inhibit the growth of the typical spoilage bacteria such as Pseudomonas and Shewanella. potential spoilage bacteria are among the psychrophilic and psychrotrophic flora present on temperate-water fish before death (9). In the absence of oxygen. What is more. often in the absence of adequate refrigeration. By vacuum packing foods. oxygen etc. Packaging changes the intrinsic and extrinsic parameters affecting a product. Low oxygen levels have been shown to substantially prolong the freshness and quality life of refrigerated seafood products. nitrogen. the natural atmosphere is replaced with a controlled gas mixture (carbon dioxide. they are usually process contaminants. partial fermentation of sugars occur leading to lower pH. allowing more growth of spoilage organisms. by actively altering the composition of the 6 .2 Modified-atmosphere packaging A natural atmosphere rich in oxygen (21%) is responsible for oxidative processes and for all aerobic respiratory life.3. if there is some oxygen demand from the product. for example. Monitoring of the seawater for algal growth in order to limit the risk of algal toxin ingestion. Carbon dioxide is the most important gas in MAP of fish because of its bacteriostatic and fungistatic properties. and specific spoilage organisms may use other terminal electron acceptors naturally present in the fish (trimethylamine-N-oxide (TMAO). As with all foods. Physical barriers not only protect from physical damage. from water activity (aw) through to physical damage. However. not part of the normal flora of the meat animals (8).FISH AND SEAFOOD can be substantial. 7). but if applied properly should extend the life of the product. and of the quality of the water used for ice and to wash fish. but isolate the food in an environment different from the bulk atmosphere. careful and sanitary handling during processing is required to reduce the risk of contamination with potential human pathogens. use of effective detergents and disinfectants.). and to limit the loss of quality (5). MAP extends the shelf life of most fishery products by inhibiting bacterial growth and autoxidation. Good Manufacturing Practice (GMP) and control of the sanitary conditions of the transport and processing environments are essential to limit additional risk of disease caused by fish consumption (6. Anaerobic atmospheres have less effect on fresh fish shelf life. In MAP.
and the MAP of fish has more challenges to overcome as a result of: a comparatively large initial load of bacteria present. gutted or filleted fish. In the United States. an inert and odourless gas. which provides guidance relating to vacuum or modified-atmosphere packaging for specific fish products (14). Growth and toxin production have been detected in artificially contaminated packs of whole trout after 1 week’s incubation at 10 °C (13). does inhibit some aerobic bacteria and reduce the rate of oxidative rancidity. Modified-atmosphere preservation of fish was first reported in the 1930s. including salt. without overt signs of spoilage. slower-growing flora (11). but residual oxygen and other electron acceptors may be sufficient to allow oxidative spoilage of fish (12). nitrous oxide and carbon monoxide have also been suggested as possible replacement gases in trace amounts for MAP/CAP. and the problems of muscle structure damage by the modified-atmosphere (11). phosphates.CHILLED AND FROZEN RAW FISH surrounding gas. the higher pH and reduction potential (Eh) of the fish muscle. may not be considered ‘fresh’ fish. The principal effect of raised carbon dioxide-MAP is an extension of the 'lag' phase of the growth of the bacteria on the fish. the pathogens that may be able to grow before spoilage occurs. The microflora of meat is not the same as that of whole. VP is one of the oldest forms of altering the interior gaseous environment of a pack. discolour pigmented fish (11). after additive application. The (International) Codex Committee has published a Code of Practice for Fish and Fishery Products (CAC/RCP 52-2003). Gases used in MAP of fish most commonly include carbon dioxide and nitrogen. and increase in-pack drip (12). High concentrations of carbon dioxide have the most pronounced microbial effect. but can dissolve into fish liquids and deform packages. the National Advisory Committee on Microbiological Criteria for Foods (NACMCF) has published a number of recommendations on 7 .3 °C and produce toxins. Micrococcus and Moraxella). but only in recent years has it seen a marked expansion in use and market share. the inhibition of common 'spoilage' bacteria (Pseudomonas. as they are able to grow at temperatures as low as 3. Sulphur dioxide. MAP has been applied to fresh meat and fish with a resulting commercially viable extension in shelf life (10). Replacement of oxygen with nitrogen. sorbates and chelating agents such as ethylenediaminetetraacetic acid (EDTA) (11). This has been driven partly by increased consumer demand for fresh and chilled convenience foods containing fewer chemical preservatives. and the promotion of a predominantly Gram-positive. The products. Flavobacterium. The single most important concern with respect to the use of MAP is the potential for outgrowth and toxin production by C. although less information on their effectiveness is available. Many additives have been tried in conjunction with changed atmosphere. a modified-atmosphere may contain any gas necessary for the desired effect. botulinum. which are able to grow rapidly at low temperature. Of particular concern are the psychrotrophic type E and non-proteolytic type B and F strains.
3. It was also noted that organoleptic spoilage and rejection by the consumer should occur before the possibility of toxin production. The 1985 British Guidelines on MAP/CAP fish from the National Fisheries Institute state that: 1. The growth of these bacteria is reduced by high carbon dioxide concentrations but. The addition of high levels of carbon dioxide to these fish. at all times. 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. These conditions include storage of the product at ≤ 3. Subsequent storage must be at or below 3 °C. but is recognised as a drawback of carbon dioxide application.g.g. however.3 °C at all points from packaging onwards. oxidative rancidity is a significant source of spoilage and hence the complete removal of oxygen and the use of oxygen-impermeable films are preferred. In the United Kingdom. 5. TMAO) that can be reduced to produce 8 . For other less fatty marine fish. For fatty fish. this problem occurs to a lesser extent with meat. Freshwater fish have different intrinsic bacterial flora and also do not contain such high concentrations of amines (e. as already stated. adequate product labelling in respect of storage temperature. The problem of water loss in MAP fish is common. 2. may result in unacceptable colour and textural change and hence little shelf life extension (11). The only processing dip to be used is 5% potassium sorbate and 10% sodium tripolyphosphate. Only the highest-quality fish. Sensitive and accurate time-temperature indicators must be used to ensure that the product has not been temperature-abused. use of high-quality raw fish. The replacement gas should contain at least 40% carbon dioxide. adequate shelf life and cooking requirements. It highlights temperature control as the primary preventive measure against the possible hazard of toxin production by C. should be used. Storage studies have shown that the extension in shelf life of MAP cod fillets was proportional to carbon dioxide concentration up to around 50% (V/V). owing to the dissolution of cellular structures. the Sea Fish Industry Authority has published guidelines for the handling of fish packaged in a controlled-atmosphere (16). and the use of a HACCP plan. 4. greater advantages are seen. leading to the recommendation that the sale of MAP/VP raw fishery products be allowed only when certain conditions are met. however. Although all fish species need to be treated separately. botulinum. from where drip loss and gaping of the muscle meant that organoleptically classified rejection occurred sooner than with no carbon dioxide addition (12). rejection due to alteration in fish texture with greater carbon dioxide makes this unsuitable (13).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. from both a microbiological and a chemical standpoint. trimethylamine and sulfides).
1. and the flesh turns from bright and elastic to dull and soft (1). as traditional spoilage patterns of the external organs are typically very distinct. Gas mixtures of 40% CO2/30% N2/30% O2 for white fish and 40 .60% carbon dioxide with a balance of nitrogen for fatty fish have been recommended (16. C (C=reject) Significantly. smell and 9 . 18) and are probably the most widely used. A. and metabolic activities of microorganisms present in the fish. or for that matter evaluation of the taste of the fish.III. with no smell. loss of the fresh characteristics may be simply measured by comparative visual and smell analysis.4. Spoilage of fish starts upon death due to autoxidation (oxidation of unsaturated lipids).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. rejection of whole fresh fish for human consumption may be made without chemical or microbiological evaluation. Various methods for whole fish freshness evaluation by a trained panel have been proposed. Over time. B. Taste panel evaluation of cooked fish may be used to score or grade colour. The eyes turn from convex and clear to concave and opaque. the gills from pink and shiny.4 1.1 Spoilage Fresh fish spoilage and methods of evaluation Food spoilage can be considered as any change that renders the product unacceptable for human consumption (10). TABLE 1. reactions caused by activities of the fish’s own enzymes. these are summarised in Table 1.CHILLED AND FROZEN RAW FISH off-odours. 17. so lower carbon dioxide concentrations may have better preservative effects. texture. but it is a commonly held view that loss of freshness is related to autolytic degradation and spoilage is more microbial in origin (1). When working with fillets. and this pattern is the basis of freshness grading schemes. evaluation of taste is unavoidable as the visual pattern of degradation of the eyes and gills is unavailable for analysis. the skin turns from iridescent to dull and bleached with bacterial slime. to brown and slimy with an intense off-odour. Degradation of whole fresh fish stored in ice generally follows a set pattern. Loss of freshness and spoilage cannot be separated as processes.
chemical. %). a ratio of concentration of the hypoxanthine (Hx) and inosine (HxR) to total concentration (ATP. HxR and Hx) gives a quantity (K-value.5 (18. TMAO permits a high Eh to remain in the muscle tissue. From being flaccid. ii) Neutral taste. as little endogenous reduction of TMAO occurs (18). AMP. post-rigor values are typically in the range of pH 5. RT Freshmeter or Fishtester). biochemical and microbiological actions. an intense odour compound.g. the Torrymeter. thus. an anti-freeze. By measuring the concentrations of the six components. ammoniacal. spoiled and putrid. ADP. the Eh of fish muscle remains relatively high (20). 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. These processes are strongly influenced by the physical conditions of storage. the conductance generally increases. The processes of degradation being analysed by the quality scoring methods above are a complex mix of physical. development of sickly sweet. or its remaining shelf life (1). To a varying extent. the muscles relax again and the fish returns to a flaccid state. After death. possibly slightly metallic. The process of ATP breakdown is used as an indicator of fish freshness (24). the pH drop of the muscle is correspondingly less. rigor resolves.8 to 6. After death. sea-weedy taste. the muscles harden as residual adenosine triphosphate (ATP) is reduced and the myosin and actin filaments bind to form actomyosin (18). reduction in ATP and increase 10 . this will generally correlate with the length and severity of rigor. or simply a waste product present due to bioaccumulation (22). the cooked analysis of fish passes through four phases of spoilage: i) Delicate sweet. fruity and/or bitter off-flavours. However. After some time. sulphurous and/or rancid smells. 19). which has been ascribed a number of possible functions as: a trimethylamine (TMA) detoxified waste product. During the initial storage. 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. Because fish tend to have relatively little residual glycogen compared with mammals. bacterial reduction of TMAO to TMA. all marine fish use/have TMAO (21).FISH AND SEAFOOD taste of cooked fish. Generally. 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. rigor mortis is the first noticeable change in the fish. 23). which increases from 0% towards 100% with time. iv) Enhancement of the spoilage characteristic of phase iii. is significant and may even be responsible for the ultimate sensory rejection of fresh cod and other fish with high initial TMAO content (20. As the fish degrades. iii) Traces of sour. cabbage-like. an osmoregulator. IMP. little flavour. texture becomes soft and watery or hard and dry.
which may also be formed by bacteria (25). or a specific fraction of the volatile bases. not to describe freshness. a correlation can be made between the spoilage of the fresh fish and the production of TVB-N. Neither does the change in pH during storage correlate well with the production of TVB-N. During spoilage. hypoxanthine. Chemical degradation continues after the initial post mortem phase. so the measurement is effective only during the initial loss of freshness. therefore. 31). colour changes. as microbial spoilage is limited by the low storage temperature (1. the residual acid is then titrated. and are especially important in the spoilage of frozen fish. The variation in post mortem fish pH may likewise influence the amount of bases being liberated to the air and consequently affect the odour characteristics of the fish. However. The Conway method (and variations of it) uses a strong inorganic base to volatilise the bases in the fish sample. 26). Comparing results for different fish species. does not show correlation between muscle pH and the amount of volatile bases contained within the fish at rejection.CHILLED AND FROZEN RAW FISH in hypoxanthine by endogenous enzymes allow this measurement to be used for determining freshness (19). This measure is highly dependent upon the temperature of storage and may not reflect the rate of loss of quality equally at different temperatures. however. Volatile bases are the best-characterised chemical indicators of fresh fish spoilage. also termed TVN). Evaluation of Total Volatile Base-Nitrogen (TVB-N. Oxidative rancidity is known to reduce the quality of fatty fish in particular (19. Quality indices based upon the products of microbial metabolism do not explain changes in quality until microbial growth produces measurable changes in the fish. as significant proteolysis is observed only during the latest stages of spoilage and after rejection (30. using Conway diffusion chambers (29) allows determination of changes of mg-N/100 g fish. the importance of microbial action increases with time (5. Lipid oxidation can be a result of enzymic action or a cascade reaction initiated by free radicals (26. Lipid oxidation and other oxidative changes lead to oxidative rancidity. and a segregated weak acid to absorb them. is later significantly reduced by bacterial action. 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. however. The major spoilage odours and flavours of fresh fish are undoubtedly principally microbial in origin. For some fish species. 7). 11 . these measurements are usually used to quantify the amount of spoilage. for example the TMA fraction. 27) produced by aerobic respiration and other forms of metal ion reduction (28). the majority of volatile bases are produced from the soluble non-protein nitrogen of the fish (free amino acids and other low-molecular-weight nitrogenous compounds).
The concept of specific spoilage organisms (SSOs) is not new. This increase has been reported to be moderately dependent on the temperature of the sea from which the fish are taken (33. biochemical and microbiological reactions taking place within and on the fish. a minority are often associated with the majority of the spoilage. 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.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. Not all the bacteria growing on a food will lead to the production of objectionable characteristics. Fish spoil as a result of the chemical. the pH of the fish muscle may change (intrinsic) and a different microbial population may develop due to the change in atmosphere and pH.FISH AND SEAFOOD 1. the lower the temperature of storage.5. The reasons for the differences between tropical. For example. For fresh fish. careful temperature control during storage is not only important in terms of quality loss. but also crucial for the assurance of consumer safety. 34).5.5. 1. The sum of these changes may result in a different spoilage profile. the results of altering one parameter may have far-reaching consequences on others. realisation that the bulk of the microbial population on newly caught fish does not cause off-flavours and off-odours stems from work 12 .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). 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). including the ability of the endogenous bacteria to grow at low temperatures and lower endogenous enzyme activity. For fresh fish iced immediately after capture and continually stored in ice. the slower the spoilage processes proceed (within limits).5 1. Also. 5). 1. The factors are interactive and cannot be completely isolated. The application of ice storage increases the shelf life of fresh fish from a matter of hours at ambient temperatures to days or weeks.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.
Sh. motile with polar flagella.g.5. oxidase-positive.3. therefore. and may play some role in the spoilage of freshwater fish. The spoilage compounds associated with the growth of psychrotrophic Pseudomonas spp. During the 1970s. 35. more especially. It is thus an important spoilage organism of gadoid fish such as cod. the spoilage of a product may be strongly influenced by the conditions under which the product is held. obligate respiratory bacteria. it is often isolated as about 1 .5. its ability to reduce TMAO to TMA. The importance of Sh. 1.10% of the total flora of fresh fish from temperate marine waters. work with inoculated sterile fish blocks resulted in identification of a specific minority of microorganisms. form methylmercaptane (CH3SH) and dimethylsuphide ((CH3)2S) from methionine and produce hypoxanthine (Hx) from inosine monophosphate (IMP) or inosine.CHILLED AND FROZEN RAW FISH started in the 1940s. They are generally characterised as Gram-negative rods. putrefaciens to the spoilage of fresh marine fish has been recognised since the 1940s. esters and nonhydrogen sulphide sulphur-containing compounds such as methyl sulphide (36. produce hydrogen sulphide (H2S) from cysteine. i. or faster than the rest of the flora of ice-stored fresh marine fish (9). putrefaciens spoilage of fish is due to its biochemical action on muscle. which produced the characteristic spoilage compounds of the fish (30. Pseudomonas spp. ‘oniony’ and ‘faecal’ odours from the production of ketones. although only since the late 1960s has it been realised that specific metabolites of Sh.e. frozen) fish products. aldehydes. 13 . putrefaciens is considered an SSO of temperate-water marine fish species stored in ice. the characterisation of spoilage of each product must be made before the identification of the responsible agent(s) can proceed. putrefaciens growth may be used as indicators of spoilage (30). As previously noted. (9) and they are considered SSO of iced freshwater fish (35). on fish are diverse and in many cases species-specific. and proteolytic and lipolytic enzymes that may affect the quality of fresh and. 37). It is able to grow as fast as.1 Pseudomonas spp. It is also present in fresh water. catalase-positive.2 Shewanella putrefaciens Sh.3. mediated spoilage is characterised by ‘fruity’. Members of the genus are able to produce pigments. These organisms are described as potential spoilers. 36). processed (e. 35). The spoilage of freshwater fish is generally ascribed to the growth of Pseudomonas spp. The Pseudomonadaceae family represent a large and poorly defined group of microorganisms. plus other characteristic compounds of the species responsible for spoilage. 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. Bacteria identified as being associated with the spoilage process of fresh fish are as follows: 1.
putrefaciens growth has been collected (12. This wide-ranging respiratory capability is thought to be unique (39). 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. putrefaciens has been studied. the bacteria have a significant problem to overcome . whereas >108 cfu/g Sh.3 Photobacterium phosphoreum Recognised for some time as being present on spoiling fish (30). unlike many other motile bacteria.5.FISH AND SEAFOOD for which the most compelling evidence showing spoilage as a result of Sh. although. Shewanella gelidimarina.3. This type of intoxication occurs when bacteria convert the histidine 14 . 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). Ph. mesophilic strains of Sh. it is strictly respiratory. as the ferric iron is insoluble. a population of Ph. Shewanella affinis. In terms of its taxonomy. baltica has been identified as the main H2S producer in cod during cold storing (37). Another distinct feature of the physiology of Sh. Shewanella waksmanii. and Shewanella aquimarina. putrefaciens were required. Sh. In 50% N2/50% CO2 MAP cod. Ph. The prevalence of the bacterium in so many environments.how to reduce a molecule that they are not able to take into their cells efficiently. putrefaciens lies in its chemotaxis: the bacterium. The localisation of the reductase on the outer cellular membrane offers the bacteria a solution to this problem (40). additional psychrotrophic species such as Shewanalla baltica. Because no other TMAO-reducing bacteria were present in sufficient numbers to produce the quantities of TMA that were related to rejection. at the time of rejection. putrefaciens may grow in the absence of oxygen using alternative terminal electron acceptors (ATECs). they are heterogeneous. Sh. 41). but Sh. with several new species being described. Sh. it was proposed that this organism. like all members of the Pseudomonadaceae. does not show carbon-source chemotaxis. owing to its cell size and activity. Shewanella olleyana. In fact. Shewanella livingstonensis. Shewanella oneidensis. but does show strong chemotaxis up gradients of most of their alternative electron acceptors (39). phosphoreum has also been identified as responsible for histamine fish poisoning. 1. and its metal ion reduction ability has led to intensive investigation into its role in iron and sulphur cycles. The ferric reductase activity of Sh. Later. putrefaciens were identified as Shewanella algae. Research has shown that approximately 107 cfu/g of Ph. putrefaciens was not present in such numbers (23. 41). Shewanella frigidimarina. Initially. and recent taxonomic description using modern molecular methods bifurcated this species. phosphoreum sufficient to cause spoilage was found. Shewanella denitrificans. phosphoreum were required for 50% of taste panellists to reject a sample. 38). and Shewanella profunda were described. putrefaciens strains are characterised as microaerophiles or anaerobes.
but with proper cooking. types B. botulinum recorded up to 1963. 1. age. 5) reports that fresh fish consumption has never been shown to cause human botulism. 75% (with a 35% mortality rate) occurred in Japan. where a particular 15 . phosphoreum strains have great capacity as histamine producers even under refrigeration conditions. E.1 Clostridium botulinum C. The remaining few bacteria present a major risk involved with the consumption of raw seafood such as sushi or oysters. Of these. (and very rarely F and G) have been reported to cause human disease. especially type E botulinum intoxication. 1.6. Review of outbreak data suggests fresh fish to be safe. Acetate production has been reported as a good indicator of spoilage by this organism. E. 5). they do not produce significant product spoilage). However. 1. Types A. human viruses. B.3.CHILLED AND FROZEN RAW FISH present in fish into histamine. Recent studies have investigated the dominance of B. 5. thermosphacta in the spoilage of some MAP fish. thermosphacta is a well-characterised psychrophilic spoilage organism of meat.4 Brochothrix thermosphacta and lactic acid bacteria B. thermosphacta did not produce off-flavours until 2 . it is bacterial risks that increase after capture of the fish. MAP studies have also demonstrated this organism’s sensitivity to oxygen and have shown that they are also inhibited by high CO2 concentration (43). botulinum. these risks are substantially reduced (1. Adult disease is caused by the production of the botulinum toxin. thermosphacta on spoiling fish in a 40% CO2/30% N2/30% O2 MAP (42). Huss (1. The heterogeneity of the types (A. this is probably due to spoilage occurring before toxin elaboration. E and F include bacteria with minimum growth temperatures of approximately 3. a convenient but diverse species of bacterium with a number of different types. a neurotoxin that causes flaccid paralysis and is associated with a variable (44) mortality depending upon dose. B. Fewer still are capable of growing on fresh fish. B. Some Ph. previous exposure and access to supportive treatment. algal toxins. When inoculated into VP corned beef and sliced ham. bacteria and higher parasites all present some risk associated with fish consumption (1.3 days after having reached 108 cfu/g (42). 6).e. C1. C2.3 °C (Group II) and these are non-proteolytic (i.6 Pathogens: Growth and Survival Endogenous chemicals. Growing evidence suggests a role for B. D. lightly preserved fish products are associated with botulism. including antisera. There are few human bacterial pathogens that can cause primary infections or disease and are capable of persisting in the aquatic environment. presents a potential risk.5. Of 404 intoxication outbreaks of type E C. F and G) is well documented (44).
sushi) (5). storage and preparation (11). Currently.6. vulnificus is associated with warm marine and estuarine waters. use of post harvest treatments and/or cooking of fresh seafood will reduce risks. there remains only an ‘extremely small’ risk of intoxication if reasonable precautions are taken during handling. due to the cultural preference for raw fish dishes. This strain has been responsible for 50 to 80% of all V. is small. parahaemolyticus infections since 1996 and is referred to as the pandemic strain (47). Human disease caused by the organism has only been observed in conjunction with marine bivalve (mostly oysters) and some crustacean consumption. again. but as fresh fish are often cooked before consumption and the botulinum toxin is heat-labile. 1978 . I-sushi. The bacterium causes primary septicaemia.FISH AND SEAFOOD fermented raw fish product that is consumed uncooked. the risk posed to the consumer of fresh fish. Japan. Only about 1% of marine isolates produce a thermostable haemolysin. V.g.2005). France. parahaemolyticus in sea water fish and shellfish that could potentially be hazardous to humans. whether stored aerobically or in modified-atmospheres. smoked. Generally. V. or cooking at 80 °C for 5 minutes) (44) will substantially reduce risk. a Japanese boiled and semi-dried sardine dish. USA. especially in individuals with underlying diseases (patients suffer from immune and liver diseases or blood 16 . were associated with finfish between 1978 and 1998 (46). Since 1996. parahaemolyticus is a marine organism.g. Other lightly or semi-preserved (e. parahaemolyticus O3:K6 is responsible for many of the recent V. The first report of the organism being a foodborne agent was with shirasu. salted or pickled) fish have also been associated with botulism. 5). Southeast Asia. parahaemolyticus outbreaks. the majority of outbreaks are caused by consumption of raw fish products (e. the majority are associated with products not cooked immediately before consumption. and North America.2 Vibrio parahaemolyticus and other vibrios V. Russia. there is no specific guideline that describes a minimum level of V. and proper cooking (boiling for 1 minute. accounted for the majority of cases (1. and when in contact with contaminated water (no CDC reported cases from fish. which can cause human gastroenteritis. which is believed to be required for virulence. which was probably contaminated from an uncooked food source or an excreting foodhandler. 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. including epidemics in India. 1. A unique clone of V. In Japan. 11). parahaemolyticus cases have increased across the world. The risk of toxin formation before apparent spoilage has been studied for some MAP fish (45).3 °C) will eliminate all risk. Proper temperature control (<3. Despite the ubiquitous nature of type E C. only shellfish are associated with the disease and no reported cases by the Center for Diseases Control. Proper chilling. 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.
It is particularly associated with disease following consumption of raw oysters from warm sewagepolluted waters. They are aquatic organisms with an epidemiology that is yet to be fully understood. Again.3 Aeromonas Aeromonas spp. monocytogenes has been well documented as a foodborne human pathogen. and especially estuarine environment.CHILLED AND FROZEN RAW FISH disorders). and fish and shellfish are probably the major reservoirs for the organism (50). shigelloides are encountered. 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. in its habitat. but proper cooking should substantially reduce risk of infection. coli on isolation media (50).4 Listeria monocytogenes L.. Chill storage may not eliminate growth. cytotoxic. with minimum growth temperatures around 0 °C and optima of 15 . There is no risk associated with the consumption of properly chilled and cooked fresh fish. are associated with human diarrhoeal illnesses.26 to 0. proper chill storage will eliminate any risk associated with this organism. Despite this. shigelloides is similar to Aeromonas spp. L. P. though it is a true mesophile with a minimum growth temperature of 8 °C and shows consequential seasonal variation in its environmental isolation. however.20 °C. monocytogenes incidence has been between 0. The bacterium has been documented as being responsible for a few fishborne outbreaks of gastroenteritis. 1. 1. others are psychrotrophic mesophiles (49). there is no risk from the consumption of properly handled and cooked fresh fish. Similar problems as those encountered for Aeromonas spp. but the bacterium is known to survive and grow in the shallow marine. infection. Since 2000. It is known that Aeromonas spp. is historically associated with faecally contaminated water. 17). Although the psychrophiles are unable to grow at body temperature. no more than circumstantial evidence exists linking the consumption of seafood to Aeromonas spp. possibly due to under-reporting because of their likeness to E.6. the causative agent of cholera.55 cases per 100. listeriosis cases have been reportable to the CDC. cholerae O1. which is often fatal (>50%). 5. and especially Aeromonas hydrophila. Some isolates are true psychrophiles. are present on fish and can grow to significant numbers during storage (1. sodium channel blocking and haemolytic activity (48). and proper cooking will substantially reduce risk. V. Most cases have been sporadic rather than associated to large outbreaks. In the US. in assessing the risk of fishborne infection by P. their potential to produce disease-causing exotoxin in food has not been fully examined (48). The major virulence factor appears to be the production of toxins with enterotoxic.000 17 .6.
Like other toxins.6. but cooking will significantly reduce the risk of infection. there is a possibility for growth of the organism on chill-stored fish. there is a far greater risk of unacceptable histamine levels being attained before sensory rejection. 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. 1. it is not apparent to the consumer and it cannot be destroyed by cooking. but more especially by Morganella morganii. some Vibrio spp. alcoholics. D60 of 1. Decarboxylation of histidine by a wide range of bacteria including Enterobacteriacae. It is environmentally ubiquitous but its true frequency in the marine environment is not well studied. and Lactobacillus spp.FISH AND SEAFOOD persons. within high histidine-containing fish (especially members of the Scombridae and Scomberesocidae families) is responsible for an intoxication termed scombroid poisoning (54). 1. Low-temperature storage of potentially toxigenic (<5 °C) fish at all times is the most effective way to control histamine production (6). Photobacterium spp. Infections are mostly associated with the consumption of raw or 18 .5 Scombroid fish poisoning The production of histamine and other biogenic amines with human immunological activity. If fish are not properly refrigerated. 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. Owing to the dependence of the virulence of this organism on the immune state of the host (usually requiring lowered cellular immunity..6 Parasites Helminthic parasites can occur extensively in finfish but very few are capable of infecting humans. especially pickled fish. at risk groups . but proper cooking will reduce any risk.. scombroid poisoning is usually a mild intoxication and it is not a significant cause of death.pregnant women.98 minutes in cod and D60 of 4. leads to the production of histamine. foetuses. Fortunately. 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. Clostridium spp. It is regularly isolated from seafoods (51). full risk analysis is not possible (52).48 minutes in salmon have been reported (53). 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. Lightly preserved products.2002 (55)) and is also common worldwide. Being a psychrotroph. AIDS patients and patients undergoing immunosuppressive therapy) and a lack of data covering infectious doses. Klebsiella pneumoniae and Hafnia alvei. neonates..6. The organism poses a serious risk in chilled products not cooked before consumption. Of fish-transmitted human diseases scombroid poisoning is commonly reported in the US (118 outbreaks with 463 cases reported by CDC 1998 .
. 1997) Huss H. TABLE1. 1. FAO. Ababouch L. Faecal coliform counts may be used instead of E. which in many cases will eliminate the risk. 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. especially in warm-water areas and where fish are to be consumed raw.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.. parahaemolyticus Staph. it may be desirable to test for Salmonella and V. risk of human disease caused by natural bacterial contamination (not through faecal pollution) of fresh fish is extremely low. 2003. coli counts where this method is preferred.7 Published Microbiological Criteria Microbiological criteria for food defines the acceptability of a product or a food lot based on the absence or presence or number of microorganisms. area or lot (CAC. Rome. Criteria should set standards that are attainable by the currently accepted GMP and applied only when there is absolute need for it. The most effective control of all bacterial risks is continual chill storage in ice (0 °C) or freezing. 444. No. Testing methods should be practical and the enforcement of such criteria should translate as a reduction of potential microbiological risks to consumers. A limited number of nematodes and trematodes found in finfish have also been identified as a cause of human disease (56). An increase of APC to levels in excess of 106 cfu/g is usually indicative of inadequate refrigeration. parahaemolyticus (see Table 1. Gram L. Cooking properly immediately before consumption is also an effective way of reducing or eliminating risk of fresh fishborne disease. long storage under refrigeration or one of the former prior to freezing.H. In summary. coli proposed by the International Commission on Microbiological Specifications for Foods (ICMSF).IV) (57). Assessment and management of seafood safety and quality FAO Fisheries Technical Paper. aureus 10 7 7 2 3 3 19 5 5 5 0 2 2 0 102 103 103 104 . 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. volume. EC. For fish from inshore or inland waters of doubtful microbiological quality.CHILLED AND FROZEN RAW FISH mildly processed fish such as sushi.1997. including parasites and/or quantity of their toxins / metabolites per unit of mass.
The role of seafood in bacterial foodborne diseases.. Armstrong D. International Journal of Food Microbiology. Rhodehamel E. Kirby R. Gibbs P. M. Huss H.H. Simpson B..E. 121-37. International Journal of Food Microbiology. 9. Rome.. Jehanno D. Nychas G. 87-118. Code of Practice for Fish and Fishery Products (CAC/RCP 52-2003)... Shelf life extension and safety concerns about fresh fishery products packaged under modified atmospheres: a review. 2003. 19. References Huss H. Kautter D. Davies A. 1993. Journal of Food Safety. 1996. Gram L. 20 ... 8 (2).H. Incidence and growth of psychrotrophic Clostridium botulinum in foods. Italy. 2. 11. Smith J. Microbes and Infection. Jeksrud W.A.A. Control of indigenous pathogenic bacterial in seafood. Review: Assessing and managing risk due to consumption of seafood contaminated with microorganisms. 1994. 27. International Journal of Food Science and Technology.N. 1996. 348.R.. in FAO Fisheries Technical Paper No. activities and safety.H.J.. Sivertsvik M. A review of modified atmosphere packaging of fish and fishery products – significance of microbial growth. Food and Agriculture Organisation. 4. 291-304.A. Ahmed F. Davies A. Codex Alimentarius Commission. 24. 334. 12.E.H. Food Control..J. 243-60.J. 33. 107-27. Food Control. and natural toxins. 283-94. Ed. Food Control. 1995.8 1. Reddy N.H. Spoilage and shelf life of cod fillets packed in vacuum or modified atmospheres. 7. Quality and quality changes in fresh fish. Fletcher R. 2002. Capell C. Ashie I. International Journal of Food Sciences and Techonology.T. 1992. 1994. Dalgaard P. Assurance of seafood quality.K. Food and Agriculture Organisation of the United Nations. 2000. Gram L.R. 91-8.. 1989. Codex Alimentarius Commission.R. 1992. Food and Agriculture Organisation. parasites.FISH AND SEAFOOD 1. 12.S. Rosnes J. Food and Agriculture Organisation of the United Nations. 87121.P.. Theoretical analysis of two hot water cabinet systems for decontamination of sides of beef.. Incidence of foodborne pathogens in European fish. 2001. International Journal of Food Microbiology. Spoilage and shelf life extension of fresh fish and shell-fish. 37. 1997. 13. 36. Ed. Microbiological spoilage of fish and fish products. Davey K. 14. Huss H. Critical Reviews in Food Science and Nutrition. 5. 10..R. 12. Huss H. in FAO Fisheries Technical Paper No. 5 (1). 67-71. 1651-60. Huss H. 3... 8. 5-7. Feldhusen F. 6. Rome. 2.K.
International Journal of Food Microbiology. Martin R.. 21. 29. Guidelines for the Handling of Fish Packed in a Controlled Atmosphere.. Davies A. 36. Gram L. in Chemistry and Biology of Marine Food Products. 121-37. 41-50. Chemical composition. 11.A. Food and Agricultural Organisation of the United Nations. Flick G. Edinburgh. Qualitative and quantitative characterisation of spoilage bacteria from packed fish.CHILLED AND FROZEN RAW FISH 15. Vacuum or Modified Atmosphere Packaging for Refrigerated Raw Fishery Products. Microbial and biochemical spoilage of foods: an overview.A. 27. The micro-determination of ammonia.E. Leblanc P.M. 26. Rome.edu/haccp/compendium/chapt08... Byrne A. Hendrie M. Yajima T.G. 1988. 1995. Flick G. Lancaster.H. Occurrence and significance of trimethylamine oxide and its derivatives in fish and shellfish.ucdavis. 1992.E. Huis in't Veld J. Huss H. Martin R.. 1933. Biochemical Journal.S. 27.. Hebard C.a review. International Journal of Food Microbiology.J. Flick G. 305-17. 1415. Herbert R. 20. Hebard C. International Journal of Food Microbiology.E..E. 2007. Journal of Agricultural and Food Chemistry.J.M. http://seafood. Arai K. 24. 1987. 1985. Saito T. Food Technology. Westport.J. Khayat A. 1983. 1971. in Advances in Seafood Biochemistry. 1992. Rome. 42-54.J... 1995. 245-67.. 419-29. Shewan J. Journal of Food Quality. Quality and Quality Changes. 130-2. Ratnayake W.. AVI Publishing. 184.J.. Conway E.M. 28.J. 117-27. 22. Gill T.. 1995. Ackman R.N. 1996. An absorption apparatus for the micro-determination of certain volatile substances. 1996. Technomic. Gill T. March 20. 23. Eds. 34. Surette M.C. Non-enzymatic oxidation of seafood lipids.spoilage of fish. Shelf life extension of fresh fish ..J. 30. Reginstein J. 1. Bacteria active in the spoilage of certain seafoods. 19-22. 1995.J. Adopted. in Fresh Fish. Huss H. Ed.H.M. 1-18.R.htm Anon. Martin R. Sea Fish Industry Authority. National Advisory Committee on Microbiological Criteria for Foods. Journal of Applied Bacteriology. Biochemical basis of post-mortem nucleotide catabolism in cod (Gadus morhua) and its relationship to spoilage. Nature. 33.. Changes in purine nucleotide of red lateral muscle of rainbow trout. 19. 1982. Letters in Applied Microbiology. 18. Ed. 26. Microbiological spoilage of fish and fish products. 22-36. 33. Børresen T.. Lipid oxidation in seafood. Huss H. National Advisory Committee on Microbiological Criteria for Foods. Food and Agricultural Organisation of the United Nations. in Fresh Fish.H. Part I . Eds. Washington D.. 17.E. . 25. 354-8. Fate of Aeromonas and Yersinia on modified-atmospherepackaged (MAP) cod and trout. Autolytic changes in fish post mortem. 37. Schwall D. Pedrosa-Menabrito A. Dalgaard P.H..E. 21. 1959. Quality and Quality Changes. 21 16. Slade A. Gibson D.
Fish spoilage bacteria – problems and solutions. and regulation. 1994. 33. Myers J.. Saffarini D. 72-80. Volatile compounds produced in sterile fish muscle (Sebastes melanops) by Pseudomonas putrefaciens. II. 22 . Ed. Annual Review of Microbiology. Nychas G. 35.R. 33. Journal of Applied Bacteriology. Vogel B.J. 18-21.eu/LexUriServ/LexUriServ. 48. 34. Do fish from tropical waters spoil less rapidly in ice than fish from temperate waters? Food Technology in Australia. Huss H. FAO Fisheries Report. 40. Nealson K.. Huss H.H.S. 42.H. 1984. Applied and Environmental Microbiology.E..M. Dixon N.. physiology. Magno-Orejana F. 61-6. 328-9. Occurrence and importance of proteolytic bacteria as spoilage indicators. 1989. Growth and activity of Shewanella putrefaciens isolated from spoiling fish. Sumner J. 44. Oxford. Italian Journal of Food Science. Gorczyca E. 39. 2002. 174. 334. 262-6. Journal of Bacteriology.. a dominant microorganism in Mediterranean fresh fish (Sparus aurata) stored under modified atmosphere. 109-36. 1992. COUNCIL REGULATION (EC) No 2406/96 of 26 November 1996 laying down common marketing standards for certain fishery products http://eurlex. in Fresh Fish. Blackwell Scientific Publications. Applied Microbiology. 67. Quality and Quality Changes.. Localisation of cytochromes to the outer membrane of anaerobically grown Shewanella putrefaciens. 9. Do Tropical Fish Keep Longer in Ice Than Temperate Fish: The Circumstantial and Definitive Approaches.M.M. 311-43.R.do?uri=CONSLEG:1996R2406:20050602:E N:PDF Sumner J. Role of proteolytic bacteria in fish spoilage. Drosinos E. Iron and manganese in anaerobic respiration: environmental significance.. Cohen D. 1989. 2005. Brady P.J. Identification of Shewanella baltica as the most important H2S-producing species during iced storage of Danish marine fish.europa.. 1982. 36.. Brochothrix thermosphacta. Jørgensen B. and an Achromobacter species.. Myers C.A. Rome. Venkateswaran K.B. Dalgaard P. Archiv für Lebensmittelhygiene. 32. 1984. 317. 6689-97. 4. 38. Pseudomonas fluorescens. 1995. 26.. Karnop G. 41. 62-70. Rome. 51-62. Satomi M.H.H. The inhibition by CO2 of the growth and metabolism of microorganisms..F. 71. 323-9.. 3429-38. Scanlan R. Processes in Pathology and Microbiology. Taussig M... 36. The effect of storage temperature on shelf life. 37. Miller A. Libbey L. Environmental Biotechnology.L. Gram L. Kell D.. 1985. 1973. 43. Lee J.FISH AND SEAFOOD 31. 13. 1996. Dalgaard P. Gram L. Food and Agricultural Organisation of the United Nations. International Journal of Food Microbiology.
H..L. 1994. Thompson S.K.N. 1997. Journal of Food Protection. International Journal of Food Microbiology..K. Heat resistence of Listeria monocytogenes in vacuum packaged pasteurised fish fillets. 2005..C.C.. Mac Rae I.P.and waterborne pathogens. International Journal of Food Microbiology. Experimental evidence for toxin production by Aeromonas hydrophila and Aeromonas sobria in a meat extract at low temperatures. Vibrio parahaemolyticus Infections in the United States. Bean N. Journal of Infectious Diseases. Embarek B.. Food Technology. Golden D.M.United States. 46.A. 2194200.. 2003. 330-6. Histamine fish poisoning revisited... Huss H. Ed.K. Slutsker L.P. 69.L. 181-8. Embarek B.. 20. Whiting R. Smith J.J. 1973-1998. Loessner M. 55 (SS10). Garcia G. Painter J... Risk of growth and toxin production by Clostridium botulinum non-proteolytic types B. PCR detection of a newly emerged pandemic Vibrio parahaemolyticus O3:K6 pathogen in pure cultures and seeded waters from the Gulf of Mexico. International Journal of Food Microbiology.M. 51 (4). 1987. Springer. Ljungh A.. 51. Altekruse S. 49. 181. Modern Food Microbiology. Ray B. Presence.. Bishop R.W. Olley J. The International Commission on Microbiological Specifications for Foods. Lynch M. 23 . 17-34. USA. 303-12. The International Commission on Microbiological Specifications for Foods. International Journal of Food Microbiology. Sampling for Microbiological Analysis: Principles and Specific Application. 53. 2006.. 55. 2000. Microorganisms in Foods 2. detection and growth of Listeria monocytogenes in seafoods: a review. Surveillance for Foodborne-Disease Outbreaks . Hammond R.. 100-3. Blackwell Scientific Publications. Miller A. 1661-6. Kluwer Academic/Plenum Publishers. Woodruff R. 52. Daniels N.. Wilson S.. 1986. 58. Majeed K. Genigeorgis C. 2005. 54. Lindroth S. 23... 50. 85-95. 1993.M. Jay J.A.J. The International Commission on Microbiological Specifications for Foods. Braden C. Morbidity and Mortality Weekly Report. 1991. 47.CHILLED AND FROZEN RAW FISH 45. MacKinnon L. Bej A. Microbial Ecology of Food Commodities. 50. 181-96. in Microorganisms in Foods 6. 48. 56. 1991.H. Myers M. International Journal of Food Microbiology. New York. Fish and Fish Products (Microorganisms in Fish Products). Panicker G. 1-37. Applied and Environmental Microbiology. Lehane L. 12. 57.... 1-34. 2000. 172-249. Wadström T.. Use of risk assessment to reduce Listeriosis incidence. 12. Griffin P.. Aeromonas and Plesiomonas as food. Oxford. 1998-2002.. E and F in salmon fillets stored under modified atmospheres at low and abused temperatures.
Hobbs G.. Gibson D. Fisheries and Food. Kitts D..A. 317-21. Villemure G. Torry Research Station. 15. Ed..)... Ogden I. Eds. Handling and processing of scad. McMeekin T.S. Do Tropical Fish Keep Longer in Ice than Temperate Fish: the Circumstantial and Definitive Approaches. The Microbiology of Seawater Fish. 73-89. Labuza T. Further Studies on Marine Fish Stored Under Modified Atmosphere Packaging. Jones Y. 1997. 1989.H. 1989.. 1984. Gibson D. Estimation of the bacteriological quality of fish by automated conductance measurements. Journal of Food Protection. 4. 317 (Suppl. Fisheries and Food. Detection of specific spoilage bacteria from fish stored at low (0 °C) and high (20 °C) temperatures. Ravn Jørgensen B..M. 1992...H. 1992. Shinano H.. Journal of Food Science. Dalgaard P.FISH AND SEAFOOD 1. 24 ..M. Ross T. 1. Huss H. Bulk storage of cod fillets and gutted cod (Gadus morhua) under carbon dioxide atmosphere. International Journal of Food Microbiology. Gram L. 1985. 127-34.9. Magno-Orejana F. Lancaster.P. Huss H. 1988. 1984.. Picard G. International Journal of Food Microbiology. 35. Academic Press. Cho Y. Torry Research Station of the Ministry of Agriculture. Food and Agriculture Organisation. Taoukis P. Akiba M. 55.1 Further Reading Processing Anon. in Torry Advisory Note No 93. 62-70. Prediction of shelf life and safety of minimally processed CAP/MAP chilled foods: a review. Changes in microflora and chemical compounds in mackerel stored by partial freezing.H. Simard R. Aberdeen. International Journal of Food Microbiology. in Fish as Food. Torry Research Station of the Ministry of Agriculture. Aberdeen. Technomic Publishers.9. 51. Microbiological quality and shelf life prediction of chilled fish.L.. Application of predictive microbiology to assure the quality and safety of fish and fish products. Bulletin of the Faculty of Fisheries (Hokkaido University). 13-32.G.M. 741-50. Florida...D. Rome. Food and Agriculture Organisation of the United Nations.. 1986.. Houston N.9 1. International Journal of Food Microbiology. 295-307.2 Spoilage Shewan J.E. Borgstrom G. Smith G. Ed. 271-85.. 65-72.. 6.1983.D. in FAO Fisheries Report No.. in Seafood Science and Technology. Mathematical Modelling Used for Evaluation and Prediction of Microbial Fish Spoilage. 1. Cann D. Sumner J. 1961. 1.C. Trolle G. Olley J. Huss H. Shahidi F. Studies on the microbiological ecology of mackerel stored by the method of partial freezing.
.3 Pathogens Condon S.. 72. Gram L.do?uri=CONSLEG:1996R2406: 20050602:EN:PDF Davies A.eu/LexUriServ/LexUriServ. Effect of culture age. 59. 137-46. Detection of specific spoilage bacteria from fish stored at low (0 °C) and high (20 °C) temperatures.L. 65-72. Applied and Environmental Microbiology. pre-incubation at low temperature and pH on the thermal resistance of Aeromonas hydrophila. Gram L. Liston J. 1993. 1987..9. Jacobsen M. 322-6. Trolle G.R. Ed.. in Quality Assurance in the Fish Industry.. Garcia M.CHILLED AND FROZEN RAW FISH Einarsson H. Amsterdam. 1. http://eur-lex. 1987. 479-88. Inhibitory effect against pathogenic and spoilage bacteria of Pseudomonas strains isolated from spoiled and fresh fish. 1992.9..H. 20023. 25 . Huss H.M.H. London. Hall G. Blackie Academic & Professional. Klausen N.J. A rapid method for detection of histamine producing bacteria..H. 1997. Sala F. Otero A. Elsevier Science. Journal of Applied Bacteriology. 5. International Journal of Food Microbiology. in Fish processing technology.. 4. 1. Modified atmosphere packaging of fish and fish products. Huss H. Huss H. Eds. International Journal of Food Microbiology. 2197-203.4 Standards and criteria Council Regulation (EC) Number 2406/96 of 26 November 1996 laying down common marketing standards for certain fishery products.K. 1992. Predicting the shelf life of cod (Gadus morhua) fillets stored in air and modified atmosphere at temperatures between -4 °C and +16 °C.europa.
are used in chilled and frozen prepared meals since many of the attributes on which quality is assessed are not present or noticeable (e. Even though heating prior to consumption will eliminate pathogenic microbes. The majority of seafood products produced require a cooking stage before consumption. smoked (cold) salmon. it does not necessarily apply to the toxins that may have been produced. but still acceptable and meeting all safety criteria.).g. clarity of eyes. 27 . products of lesser quality. Few seafood products are sold ‘ready-to-eat’. 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. This is no longer the case with many processed products now available in stores. and crustacea such as shrimp and crabs. colourings etc. seafood (with the exception of some crustaceans) remains uncooked until the point of consumption.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. The same used to be said for the difference between fresh and frozen products. or could be disguised by sauces. 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. colour of gills. Simon Derrick The Grimsby Institute Humber Seafood Institute Europarc Grimsby DN37 9TZ United Kingdom 2. Sushimi tuna). the exceptions being Sushimi (high quality raw fish).g.2. due to the early cessation of spoilage. although recent developments in improved supply chains. smell etc. This has the result that any microbes present in the product have the capability to grow (with subsequent spoilage) unless strict temperature controls are applied. CHILLED AND FROZEN PREPARED FISH PRODUCTS Dr. smoked (hot) mackerel. emphasising the high quality of the seafood and the gourmet nature of the prepared products.
and these are increasingly being presented with an ever-expanding range of accompaniments. Nevertheless. At the same time. molluscs. where the preparation has already been carried out. There is also a limited market for filleted material. the processing industry has developed a wide range of value added products. including freshwater fish. Doubts have been raised over the microbiological status and handling practices of reef-caught fish. Marketing-style descriptions and process definitions are presented below to acquaint the reader with the variety and forms of seafood available. 2. Cold-water fish microbiological specifications are inappropriate for assessing freshness of warm-water fish. Seafood is the term used descriptively to include all categories of product. Warm-water/reef-caught fish were initially supplied to add colour and variety to wet fish displays. Prepared natural fillets are fillets of fish without any additional coating. which can be farmed. and the consumer only has to heat (cook) the product. and this has produced a greater availability and diversity of products in the marketplace. prawns.. They can be with or without skin and may contain pin-bones although the major skeletal bones are removed during filleting. rather than frozen. they are usually of a better standard than species such as tilapia. This includes all categories of seafood including trout and other freshwater species. and are well known for their diet consisting of waste materials. 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. etc. The expected shelf life attainable for this format is usually up to 8 days.FISH AND SEAFOOD Recent trends in the market for food and seafood products. as they tend to lose colour and sheen after defrosting. and pathogenic Vibrio spp. Frozen fillets may have a surface layer of 28 . have shown the increasing importance of convenience foods. Listeria spp. shellfish and molluscs. It is sold from wet fish counters on ice or pre-packed in modified atmosphere packaging (MAP). but these are now finding favour as products in their own right. they are usually supplied filleted with head on. but should be equivalent to cold-water products. or in MAP depends on catch conditions and treatment.. They are typically flown into export markets. Microbiological hazards include Salmonella. Wet or chilled/frozen whole fish are whole fresh fish that may or may not have been filleted.2 Definitions The traditional focus of the chilled and frozen fish market has shifted over the last few years. from both wild capture and aquaculture sources. in particular. often using the simplicity of a microwave oven. The market spans fresh fish counter sales and frozen recipe dishes with fish as the predominant source of protein. Shelf life on ice.
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. haddock. etc. with a minimum residual salt content of 3. in addition. 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. or pre-packed frozen with added glaze. They are sold from wet fish counters. Again. Fish steaks can be cut from a natural fillet or loin (e. Fresh fillets are sold from wet fish counters. cod).CHILLED AND FROZEN PREPARED FISH PRODUCTS frozen water that forms a protective glaze. This process in effect cooks the product and reduces or eliminates microbial contaminants. a protective glaze may be added to the frozen steak. Cold smoking is essentially a flavouring process. tuna steaks) or band-sawn from a frozen block (e. The result is fish with a typical residual salt content of 1. or pre-packed in MAP or vacuum packaging (VP). They are then kilnsmoked to a minimum core temperature of 66 ºC for 2 minutes. in VP or through mail order.g. mackerel. 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 .g. 29 .2 days) longer than that of non-prepacked fish. Hot-smoked fillets are prepared by brining fish fillets in a 26% salt solution with added natural colours to provide a range of product types.g. Added colours can be included with the brine solution to meet specific requirements for products e.. A typical cold smoking process may pretreat the fillet by dipping in a 26% brine solution with a retention time of approximately 3 minutes.g.5%. and the product then shaped and heat-processed to cook the product before being chilled. and salmon. mackerel. natural fillets of fish such as herring. The expected shelf life of these products is up to 8 days (or longer for some chilled. VP products) because of the high salt content acting as a barrier to spoilage. Pâtés are prepared with raw fish. the ingredients premixed. Some products may be salted for flavour only (e. The shelf life for MAP fillets is typically up to 5 days. salmon. Cold-smoked natural fillets are prepared from a range of cold water fish including herring. and typically uses natural smoke passing over the fillets suspended on racks in a kiln (oak chippings. Hot-smoked fish are prepared from brined. which may.5 . A core temperature of around 27 30 ºC for approximately 100 minutes is a typical processing condition. resulting in a product that is ready-to-eat without any further cooking. swordfish steaks).2. The pâtés are sold from segregated ‘wet fish’ counters as a ready-to-eat product. sawdust with other woods adjusted for flavour profile and burn duration).5% in the aqueous phase. be flavoured with herb dips or peppercorns. annatto with smoked haddock. but may be increased (up to 1 month) by packaging methods like MAP or VP.
g. and may also include additional ingredients such as fresh herbs and spices. long-life seafood mixes using sorbate and benzoate as preservatives. and heat-fixed. addition of ingredients such as colours. Prawns. Fish cakes are products produced by mixing fish with potato and herbs.g. tuna. e. ‘fingers’ before being coated in batter and/ or breadcrumbs.) Raw material is typically fish fillet. it can be used as a seafood analogue. and formed into round cake shapes. Breaded fillets/steaks and battered fillets are either natural or shaped fillets. This ‘catch-all’ category includes seafood salads.g. It is often used as a protein supplement. coated in batter or breadcrumbs. and in the shell. jellied eels. 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. Scampi are tails of Dublin Bay prawn or langoustine. flavours. The protein source can be fish fillet or blocks. taramasalata. and seafood mixes. binders. With further processing. and may include molluscs or prawns that may or may not have undergone a minimal heat process during their primary processing. There is an increasing market for high value fresh or thawed fish (e.or a five-stage process. The cakes are usually coated using a three. They are pasteurised products and treated as high risk raw materials. haddock. rice or pasta.. frozen blocks or canned fish and may include molluscs or prawns that may or may not have undergone a minimal heat process. The majority are products made from various types of whitefish (cod. Natural prawns from cold or warm waters can include those peeled. Band-saw dust and fish trimmings from fishblock preparation materials may be used. salmon) coated and sold as chilled products. Surimi is the insoluble protein fraction of fish after washing and processing. e. pasta or potato. with an expected shelf life of around 7 days. which may include molluscs. mixed whitefish.FISH AND SEAFOOD Seafood recipe meal centres (frozen or chilled) are places where these products are supplied without a source of carbohydrate. rice. fish nuggets/bites with dips and seafood-based snack products. 30 . cephalopods. or cut from a preformed frozen fish blocks into shapes. Other ingredients may be natural or reconstituted. Other. etc.
see Table 2. and processing conditions such as dehydration or poor filleting practices.e. skin. For a summary of background information on the hazards associated with fisheries products and ingredients. will the saprophytic flora start to invade the surface flesh (1. Grading is based on size of fish (>25 kg). which may become a focus of contamination deep within the muscle. A primary role in fish spoilage has been assigned to this organism. and his co-workers have described a large celled microorganism found in spoiling fish and identified it as Photobacterium phosphoreum (3). Tuna that does not meet the highest of grades based on colour. In summary. handling or preparation stage. as a result there is an increase in spoilage rate thereby reducing the product shelf life. i. the microbial flora of seafood is associated with the gills.1 Initial Microflora Seafood Other chapters have discussed in some detail the indigenous spoilage. and intestines of the fish. Contamination will. storage. or visible damage from gaff marks (4). whilst gaping is also dependent on a range of other factors that effect the muscle composition.I. on the whole. Tuna caught and discarded as unsuitable for sushi or other high-risk uses is often diverted for canning. Frequent rebaiting and prompt removal of caught fish is essential to limit rapid spoilage. and ingredients added to the final product. searing or bar marking). The Food and Drug Administration (FDA) has produced guidelines for handling and processing of tuna. and pathogenic flora associated with seafood. 2). fat content. 31 . and have emphasised the origin and sustainability of the fish as key marketing points. including seasonal variation. The edible tissue of some species of fish are more susceptible to gaping than others.and turtlefriendly long lining to the less discriminatory netting.3 2. The concern is that fish may be visibly acceptable but any subsequent washing process will have little effect on removing contaminants. it may be used for other products or markets depending on the product specification. product handlers. Recent public opinion has favoured the use of dolphin. Dalgaard et al. remain on the surface of the fish fillet except where “gaping” or separation of the muscle blocks occurs.3. species (Bluefin>Bigeye>Yellowfin>Albacore). and the potentially toxic by-products of histamine-producing bacteria (5. 6). other potentially pathogenic and spoilage bacterial contaminants may be introduced from a wide range of sources including the environment. is not used for sushi or other high risk (and high value) products. sensory assessment of freshness etc. After capture and during any subsequent processing. specifying internal temperature limits for fish and for limiting temperature abuse during handling and grading (1). Only after death.CHILLED AND FROZEN PREPARED FISH PRODUCTS 2. and an increased risk from a potentially unsafe product (especially those that receive a minimal heat process.
Bacillus subtilis B. Vibrio cholerae. the effects of processing and combination or use of ingredients on identified microorganisms should also be considered with respect to both the safety and quality/shelf life of the product. Clostridium perfringens. histamine Cryptosporidium. Pseudomonas. yeasts Lactobacilli. Table 2. Listeria. raw materials. for chilled products. 32 . moulds.FISH AND SEAFOOD 2. nuts and fruit Associated microflora Lactobacilli.2 Non-seafood ingredients The range of ingredients used in the preparation of chilled and frozen prepared seafood products is very diverse. Penicillium. by the HACCP team or by a hazard and risk assessment programme. moulds Bacillus. coryneforms. mushrooms. Escherichia coli. moulds Bacillus. enteric viruses 2. Moulds Enterobacteriaceae Salmonella. and associated spoilage and pathogenic microorganisms.I provides an overview of common raw materials. moulds Bacillus Bacillus. streptococci. Clostridium Lactobacilli. toxin.4. spore-formers. Staphylococcus. Aeromonas. Enterobacteriaceae. be considered at an early stage during new product development.3. shelf life determination. Bacillus cereus. and. staphylococcal toxin. cream.1 Processing and its Effects on the Microflora Selection of raw materials/ingredients It is important that the potential microbial contaminants of all ingredients.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. B.4 2. Fusarium. and milk Cheese Water/Ice Bacillus. TABLE 2. and so are the potential microbial contaminants associated with them. 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. cereus Staphylococcus aureus. yeasts Pathogenic microflora/toxins Salmonella. pseudomonads. Shigella. Clostridium botulinum. anaerobic spore-formers. coliforms. however. protozoa and viruses B. In addition. cereus Bacillus licheniformis. intermediate processing. cereus B. Aspergillus. product assembly. Staph. cereus Carbohydrates Potato flake and powder Rice Pasta Flour Dried herbs and spices Dairy products including butter.
Band-sawn materials or fish dice are normally promptly further processed. there are a number of distinct methods used. and contamination from equipment to clean fish muscle. Where the raw materials are frozen.3 Preparation of fish component of chilled and frozen recipe dishes 2. Intermediate processing and use of chill for storage of work-in-progress will help to optimise available life of perishable components.5 illustrate basic principles in the preparation of chilled and frozen recipe dishes. 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. Rapid controlled cooling of heat-processed components will help to minimise spoilage. 2. depending on the size of the fillet 33 . Generic examples have been used to illustrate only the major points.4. as the full complexity of the processes used for specific products cannot be covered in a general text. but it also poses a risk of cross-contaminating other products undergoing tempering. this may be an incidental effect.3. and potential opportunities for growth of foodpoisoning bacteria. All have the same intention. The use of water for rapid thawing of product is not recommended since it not only rapidly thaws the product surface. the core temperature. Effects on the microflora are minimal during tempering (7).CHILLED AND FROZEN PREPARED FISH PRODUCTS Any preparation stage or process should be suited to the raw material. to evenly raise the temperature of the entire frozen product from -20 °C or lower. up to -5 °C so as to improve the cutting and handling properties during processing. as a heat process is often required for functional or organoleptic reasons.4. but will also need to ensure that the optimum shelf life can be safely obtained.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. defined holding time at a chill temperature (0 – 3 °C). Examples of tempering techniques include the use of heated air. For some components. but it is assumed that any band sawing or dice process will introduce heat from blade friction.2. Figures 2.1 .2 Raw material tempering Whenever frozen seafood is used as the raw material it is normally tempered before processing. or the use of microwave technology. line extensions and process reviews.4. It is recommended that the principles of HACCP are applied to novel processes. 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 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. The “blown” meat is graded.1. Fig.Fish Cakes). The processing of three examples types are described in Figure 2. It is then size-graded. The scampi meat (tails) is removed intact from the shell using potable water whilst still frozen. the scampi is washed in potable water to remove foreign bodies.1. 2.3.1.Scampi. with the coating heat-fixed.1B .4.1 (2. quickfrozen. by mincing and mixing with a binding agent (as described in section 2.2.7 days.2).Shaped Fillets. frozen in a nitrogen blast freezer and weighed into sacks for later processing. are placed into plastic boxes and covered in ice to preserve their condition.FISH AND SEAFOOD processed. after landing. chilled or sold chilled on defrost with an expected shelf life of 6 .1 Whole-fillet / scampi Scampi (Nephrops norvegicus). On arrival at the processing plant. Frozen cores are usually batter-enrobed. may not rise above 0 ºC during processing.4. drained and then refrozen for storage until required for further processing.3. Any broken pieces or fragments of frozen tail meat are kept for use in the production of a reformed scampi product. Individual Quick Frozen (IQF) fish fillets or steaks may also be coated using a similar process. Raw material preparation stages for chilled or frozen coated fish products 34 . 2.1A .1C . 2.
ii) Fish-filleting waste . pathogens such as Salmonella and Listeria.3. 35 .3. Spoilage of fillet pieces or trimmings will not be appreciably different from that of fillet.1.4. material will need to be transferred swiftly to a chiller. as the physical structure of the material has been destroyed. Shapes may then be cut from the individual slabs using either hydraulic shape cutters or continued cutting with band-saw type blades (e.block-sawn dust. The remaining dried ingredients are mixed with the fish.4. The fish component of the cake will. off-cuts and by-products from other processes (maybe minced) are reformatted give a final product that is still 100% fish.CHILLED AND FROZEN PREPARED FISH PRODUCTS 2. are handled and stored as any other food product.7 days.1. spoil more rapidly than the dry ingredients. Waste fish generated as a by-product of another process will always be susceptible to spoilage and water loss. 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. those that use frozen blocks of whole fillets. such as aerobic spore formers and Staphylococcus spp. with the coating heat-fixed. with an expected shelf life of 6 . Fish raw material can be obtained from a number of sources i) By-product of fish shaping or cutting/dicing process . 2. from potato flake. It is for this reason that all by-products from primary processing. once the dry components have been rehydrated. or sold as a chilled product after thawing.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. and water is added to rehydrate the components. additional spoilage organisms may be introduced. more seriously.off-cuts or belly flaps.g.. or promptly processed to avoid further quality loss. iii) Fresh/fillet/minced fish block. because of its nature. Herbs or seasonings used in the recipe may contribute additional spoilage organisms or. Waste material produced from this type of processing is often reused in the third category (fish cakes). Therefore. fish fingers). This is especially the case with block-sawn dust. The mix is shaped and formed ready for packing or for enrobing. that are to be reused for other foods. chilled. This type of material will be susceptible to contamination by Enterobacteriaceae and possibly Staphylococcus spp. or those where the trimmings. Fish cake cores may be batter-enrobed. unless it is poorly handled. quickfrozen. but.2 Shaped products This type of product falls into two categories.
from flour. and to carry flavourings where required. Over time. i.4 Battering/enrobing process Figure 2.or fivestage) frozen or chilled. If a crumb coating is required. some of this initial flora 36 . Fish products. e. perfringens could. The product then passes along a wire mesh belt through a ‘waterfall’ enrober that gives an even coating of batter to the entire product. potentially. which can occasionally be isolated from protein-handling processes. and possibly Staphylococcus spp. flour. because of the intensive handling required during processing.FISH AND SEAFOOD In all of the above examples. Batter contains water. Natural fillets d. and Enterobacteriaceae from flour. chilled. which is product-dependent and not always required. starch. Spore-forming anaerobes such as C. 2. Scampi tails are particularly susceptible to contamination with Enterobacteriaceae. rapid automated processing with the minimum of delays will minimise product spoilage by pseudomonads. Breaded or battered fish fillets are fully heat-processed before consumption.4. a. spices. a characteristic flora will develop from the batter components.. with an expected shelf life of 6 . Tempered band-sawn materials b. The function of this is to improve the adhesion of subsequent coatings. the crumb is poured onto the product followed by repeat enrobing and crumbing to complete the five-stage process.7 days. and if required.e.g. fish cakes Portions or fillets may be batter-enrobed. A large variety of product formats and types can be batter-enrobed. absorb any excess moisture on the surface of the fish portion. as well as ensuring that there is minimal contamination from human processors. such as Listeria and Vibrio spp. 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. The first stage. milk powder and spices. vibrios and other fishassociated spoilage organisms. and may include milk powder.2 illustrates the typical stages of enrobing or breading. egg. IQF/pre-formed fillets c. quickfrozen. Initially. Bacillus spp. which typically consists of flour. with the coating heat-fixed. Control of temperature limits the potential for growth of spoilage organisms and pathogenic bacteria. rusk or crumb. raising agents. rapidly multiply if strict temperature control was not maintained. is the addition of a pre-dust. lactobacilli from the milk powder and flour. or sold as a chilled product after thawing. Products are preformed and presented to the process (either three. any excess is removed with an air knife.g. e.
replaced in part by process contaminants and bacterial flora from seafood. It is advisable that some control mechanism is employed to prevent toxinforming organisms from multiplying to dangerous levels. and. This can be achieved by frequent clean-downs and adequate hygiene.CHILLED AND FROZEN PREPARED FISH PRODUCTS associated with the ingredients will be diluted out. in the case of B. dry batter mixes are all potential sources. but because batter is heat-set. cereus. The FDA has issued clear guidelines on acceptable 37 . cereus can proliferate under certain conditions. contaminants such as Enterobacteriacae can proliferate and develop during the process. The origin of contamination for the two organisms is often unclear. Fig. Flow diagram outline of a typical three. Staph. time-limitation or production run temperature control. but it is believed that poor hygiene regarding product and food handlers. and both of these can produce heat-stable toxins that can survive a batter heat-fix process.2. aureus and B.and five-stage enrobing/breading process The length of production run and temperature of the batter are key factors in deciding the balance of the dominant flora within the enrober.2. With inadequate temperature control of batter systems. this is not of great significance to the microbiological standard of the finished product.
Well-managed breading and pre-mix systems that are segregated from batter enrobers present little hazard (7). pasta or potato. but they are supplied without a source of carbohydrate e. 2. and must not show evidence of disease.2.1 ºC for no more than 3 hours is recommended (1). canned fish may be used in their preparation owing to it having wider availability and low wastage.3 illustrates vegetable processing and treatment for inclusion in chilled and frozen fish recipe dishes. to produce a specific product. Flow diagram outline for recipe dish ingredients preparation 2. Fig. Figure 2.3. as this could introduce off-flavours and reduce shelf life. The protein source can be fish fillet or block.1 Seafood recipe meal centres (frozen or chilled) These products can be defined as the central component of a meal. The vegetables must withstand processing. and may include molluscs or prawns that may or may not have undergone a minimal heat process during their primary processing. A temperature no greater than 10 ºC for 12 hours cumulatively or 21. Vegetable quality is an important characteristic often overlooked in the selection of vegetables for frozen and chilled products.5 Recipe products These products involve the combining of seafood with other food ingredients as part of a recipe.4. 38 .FISH AND SEAFOOD temperature regimes and run times permitted for batter enrobers. There are two main categories.5. rice.g.4. For example.
and protected from any post-processing contamination. coli O157. In an attempt to improve washing and reduce chlorine taint. Blanching of vegetables can be an effective means of reducing or removing surface contamination.e.CHILLED AND FROZEN PREPARED FISH PRODUCTS The products will often be presented with vegetable toppings. The majority are products made from various types of whitefish (cod. rice or pasta. chopping and mixing of material. However. This is a concern for chilled products that may have a shelf presence for up to 6 days including day of manufacture. not just as accompaniments. Although the processes are claimed to be superior to using unmodified hypochlorite for washing.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. and mixed whitefish). and reduce spoilage organisms and potential pathogens. will disperse the saprophytic flora of vegetables . any processing stage. 8) will. spore-formers. For premium products. if the process is not appropriately controlled. encourage more rapid decay and loss of structural integrity. However. coliforms and pseudomonads throughout the mix. larger diced pieces allow for easier handling and a reduction in spoilage but are less attractive to the consumer.4. Obvious signs of vegetable soft rot due to mechanical damage or infection by Erwinia spp. Salmonella typhi. colour and water will occur. 2.lactobacilli. For example. frozen block or canned fish. or breadcrumbs. (7. the use of night soil or incorrectly applied manure or sewage sludge for ground preparation may contaminate produce with a wide range of pathogens. pastry. with the accompanying release of additional nutrients by processing. Raw material is typically fish fillet. even those with pH-modified dressings. control is more onerous. with cases of foodborne disease traced to Listeria monocytogenes. they should be stored under temperature-controlled conditions.5. and may include sauce accompaniments. treated as a highrisk ingredient. The opportunity for growth of L. loss of quality. monocytogenes should be taken into account when setting shelf life or holding times of vegetable components. A number of examples of direct faecal contamination of vegetables exist in the literature (8). Washing with a biocide will at best achieve a 2 .3 log reduction in bacterial numbers on the surface of vegetables. but also as components in recipe meals. For example. all-year-round fresh vegetables are preferred. coryneforms. Once vegetables have been blanched. for example. 39 . Any assumptions regarding the application of good agricultural practice to produce should be made with caution. and may include molluscs or prawns that may or may not have undergone a minimal heat process. and E. Products must be washed thoroughly to remove pests and soil. The presentation of vegetables has a significant effect on the product quality. i. acidification of hypochlorite and chlorine dioxide have been proposed and recommended as an improved alternative. haddock.
Placing such products in a standard chill store to cool down could result in condensation. Typically. and an increased risk of contamination from L. On hydration. 40 .6. monocytogenes and other bacteria that may be present on chilled surfaces such as ceilings. For this reason all products. Potato toppings made from flake or powder will normally have little associated spoilage flora. Appropriate handling facilities or equipment will be required to protect the product. such as high-risk/low-risk barriers and dispensing and/or holding at above 63 ºC. should be chilled to <5 °C as quickly as possible using either blast chillers or other equipment specifically designed for this purpose. 13). cereus). or use barrier or hurdle principles to preclude the growth of pathogenic microorganisms. a heat process in excess of 73 ºC will be required to rehydrate this component fully. rely on the use of microbiologically sound ingredients. An incident involving Staph. Products or dried ingredients contaminated with either spores of aerobic bacteria (e.1 Sauce manufacture The variety of ingredients and components used in sauce manufacture is extremely wide. Chilled pasta is becoming more popular than dried pasta as a component in many product lines. 2. where composite meals are heat-treated. 11). can germinate and rapidly multiply.g.4. a process equivalent to at least a 5D Listeria kill will be necessary. spore-formers can proliferate if adequate control of temperature is not maintained. B.FISH AND SEAFOOD Food-poisoning incidents traced to rice and caused by B. Alternatively. 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. If the sauce contains modified starches for freeze/chill stability.4. if subjected to temperatures between 20 and 40°C.) to the product (12. and then cooled slowly allowing germination and re-growth.6 Preparation of non-fish components of chilled and frozen recipe dishes 2. cereus. cereus are well known and are still a common issue in the UK (9). especially in those products that have a vegetable component or rice accompaniments. Manufacturers of chilled products with an extended shelf life must recognise the potential for growth and spoilage of psychrotropic strains of B. aureus toxin is well documented (10. such as butter garnish with black pepper. but temperature-abused pasta can also spoil rapidly by mould growth. Sauces or marinades are added to fish to broaden appeal or to accompany components. the preparation process must ensure the eradication of all vegetative pathogens from the ingredients. with the range always expanding as new tastes and cuisines are developed. Sauces can be multi-component preparations or more simple preparations.
4.CHILLED AND FROZEN PREPARED FISH PRODUCTS Fig.5 illustrate the recipe dish ingredients preparation and assembly and packaging. that of a dairy-based sauce. Marinade-type products are becoming more popular. flour products. 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. suspended in an acidified sauce with an oil component. 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. for instance. Acidified or low-pH sauce formats using acetic acid or vinegar normally require a lesser heat process for the equivalent effect. 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. The format uses flavour compounds .2. Without barriers. spices.herbs. Figures 2. and handling of processed sauce can be less rigorous than.4 and 2. respectively. Flow diagram outline for recipe dish ingredients preparation Time limitation may minimise the potential for spoilage by process contaminants or at the dispensing unit. 41 . etc. 13).
2.4. The objective of this method is to deny the aerobic bacteria (both spoilage and pathogens) the oxygen that they need to grow. which may restrict either microbial growth or enzyme activity. The oxygen-free conditions will however allow other anaerobic bacteria such as C.7.4.1 Vacuum packing A range of fresh chilled products including salmon (smoked and fresh portions).2. so replacing the oxygen necessary for aerobic growth of spoilage microorganisms.5. generally carbon dioxide and nitrogen. and tuna loins are packed by placing the product in a plastic pouch and removing all the air.2 Modified-atmosphere packaging Originally developed for the red meat industry. the method replaces air in the sealed packaging with a defined mixture of gases. Flow diagram outline assembly and packaging of recipe dish 2. These include: 2. 42 . botulinum to grow. as it dissolves in the surface moisture to form a weak acid. Increased levels of carbon dioxide will also result in a slight decrease in the pH of the surface tissues.7.FISH AND SEAFOOD Fig.7 Effects of packaging on chilled products Products with minimal additional ingredients may use packing techniques designed to minimise microbial growth.4. unless the temperature is strictly controlled to below 5 °C throughout the supply chain.
as this organism is a common contaminant of freshwater fish species (19). The fish component of recipe dishes. At refrigeration temperatures (5 . The case for growth and contamination of products with Aeromonas is stronger. However. is typically uncooked or at best lightly processed to retain eating characteristics. Vibrionaceae will succeed (17 ). and Yersinia spp.10 ºC) Enterobacteriaceae are often found to proliferate and produce a characteristic type of spoilage that includes. and in influencing the final eating quality of the product. temperature control is important in preserving chilled integrity of ingredients. followed by moisture loss. advanced spoilage gives further changes in texture and the production of characteristic ‘off-odours’. phosphoreum. but they will also be readily adapted to the nutrient source and conditions. This is considered to be the one major difference between fish and meat products. and therefore will have a greater opportunity to dominate other spoilage flora numerically over the course of product shelf life (15). but a conclusive case for pathogenicity has yet to be presented for Aeromonas spp. Nonetheless. the slow development of ammonical and sulphuric ‘off’ odours and flavours(16). 2. Not only will these organisms have a numerical advantage over other proposed spoilage organisms.. This is also true of chill storage and the distribution chain. both organisms are capable of growth at chill temperatures. initially. by a loss of texture. have both been suggested as possible spoilage or pathogenic contaminants of chilled recipe meals (18). where the protein component is nearly always heat-processed. but little on in-pack spoilage.CHILLED AND FROZEN PREPARED FISH PRODUCTS 2. The literature has good information on spoilage of individual components of recipe meals. (20). Shewanella putrefaciens and Ph.5. in numerical terms. the microflora of interest will be mainly psychrophilic or cold-adapted (14) spoilage organisms such as Pseudomonas spp. and finally flavour loss. except for some crustacea. Beyond this. at slightly higher temperatures (10 ºC).1 Chilled storage For chilled products or chilled components of frozen products. Aeromonas spp.5 Spoilage During processing. 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. 43 . but is not limited to. Spoilage will be most readily detected in proteinaceous components of ready meals. It is not clear whether the additional contaminants from components or processing will accelerate this typical seafood-type spoilage.
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. Still. Spoilage can be either a function of bacterial growth or the microbial release of free enzymes into solution. however. However. within dairy products are well known. the shelf life of products is limited by the natural spoilage process described earlier. botulinum. For example.2 Frozen storage Maintenance of product temperature at <-18 ºC will not permit bacterial growth or enzymic spoilage. It is not unreasonable to expect that spoilage bacteria release many extracellular enzymes.3 Enzymic spoilage Post-harvest changes in the quality of fish due to enzymic activity and rigor mortis are well known and understood (10). spoilage should not be considered a purely microbial phenomenon as taints and off-flavours can develop by non-microbial means (e. a recent report examining fish gut bacteria from cold waters characterised a proteolytic enzyme. Products that are developed with an extended shelf life beyond the 6 . As a note of interest. auto-oxidation of fats/oils)... 44 . released by a pseudomonad with demonstrable activity to 0 ºC and a peak in production at 10 ºC (15). and Aerobacter spp. By retaining the intrinsic spoilage flora of seafood. 2. These have been proved to impart off-flavours.g..7 days typical of fish products/recipe meals must consider the potential for growth of non-proteolytic C. also be those that have the least effect on the viability of microorganisms (21). Pseudomonas spp. where the variety of components and ingredients used in manufacture is varied and complex. Serratia spp. freezing also has a protective role in improving the safety of raw materials by. This also ensures that any post-process bacterial contamination or spore-forming bacteria in the product will not grow. producing characteristic off-flavours.FISH AND SEAFOOD 2. 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.5. This reinforces the need for prompt processing and temperature control of ingredients and finished products. Other examples of bacterial proteolytic and lipolytic enzymes produced by Alicaligenes spp. eliminating nematodes in products that do not have a heat treatment step. The conditions and freezing process most favourable for good-quality products will. Bacillus spp. Characterisation of the enzymes has demonstrated residual activity even after a heat process that would destroy vegetative cells (24).5. for example. this should also take into account shelf life of product components used in the assembly of the final product. The type and nature of the enzymes and their activity will depend on the bacterial species.
Numerous quoted figures exist in the literature regarding cell counts required before toxin production occurs. any risk of C. Cases quoting contamination of seafood with organisms such as S. In addition. especially molluscan filter feeders. 27). air.6 Pathogens: Growth and Survival There are only two species of pathogenic bacteria that can truly be said to occur naturally in fish. and some effects related to product type and time to toxin production have been noted. have been linked with food poisoning and should also be considered. Vibrio spp. 45 . water. and Campylobacter are recorded in filter feeding molluscs such as mussels. soil.6.6. botulinum when held in favourable conditions for growth (28) i. in some cases.2 Clostridium botulinum There are reports of rapid spoilage of vegetables by C.e. Inappropriate holding times and temperatures will. or a heat shock followed by holding at a temperature suitable for growth.1 Bacillus cereus This is a common contaminant of dust. This may occur where storage under gas such as carbon dioxide is used to reduce ripening and so prolong storage. histamine intoxication. typhi. farmed or free-living crustacea (30). may be of benefit within any risk assessment for chilled and frozen prepared fish products. cereus. perfringens contaminating seafood will depend on the source of raw material. botulinum type E and Vibrio parahaemolyticus (25). with pathogenic microorganisms or viruses. spices and herbs and vegetable shoots have been reported (12.CHILLED AND FROZEN PREPARED FISH PRODUCTS 2. These are C.6. and this issue may be of greater consequence for warmwater. low oxygen or anaerobic conditions. Incidents in such diverse materials as cream. 2.3 Clostridium perfringens A study examining C. Germination of spores requires favourable conditions. Counts of between 103 . etc. are dealt with elsewhere in this book. permit rapid growth of B. along with previous discussion.104/ g should be treated with some concern (26).. A number of other Bacillus spp. Additional hazards from marine bio-toxins. 2. agricultural run-off or sewage can contaminate in-shore species. and many raw or processed foods (26). 2. potato. Therefore. 13. perfringens in seafood noted a transient population of spores in the guts of fish feeding adjacent to rivers and sewage outfalls (29). Hazards attributable to the non-seafood components are listed below and this information.
6. These include inadequate refrigeration and preparing foods far in advance of planned service. 2. at the present time. 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). Chilling may help Shigella to survive. It is unclear. will colonise factory environments in a similar fashion except for possibly Vibrio spp. aureus (35). However. 46 .6.6. growth may occur in foods (26). if any other potentially pathogenic organisms. which is not affected by any subsequent cooking process. monocytogenes as a contaminant in foods (31). and will therefore cause “food poisoning” even in a product that is cooked before consumption. An acceptance has now developed in the UK of the presence at low levels of L. infected food handlers.FISH AND SEAFOOD Vegetable raw materials are potentially contaminated through soil conditioning with animal manure.6 Staphylococcus aureus There are a number of underlying reasons for the development of Staph. indigenous or other. and this is believed to be due to a tolerance to brine used in the smoking process (32). but.5 Shigella spp. monocytogenes has been frequently isolated from fish-processing plants and is thought to be able to colonise this type of environment (32. but under the appropriate conditions. These conditions will allow the production of the toxin. L. 33) due to its ability to grow at low temperatures and form “biofilms” on surfaces. the restrictions on growth rate seen with seafood will not apply. aureus enterotoxins in foods. the possibility of a sanitiser-resistant biofilm colonising processing environments should be viewed with some concern (32). Rates of contamination with smoked seafood are much higher than those with unsmoked products. All the above points should be considered within any risk assessment of the potential for growth and toxin production by Staph. 2. monocytogenes tends to grow poorly at refrigeration temperatures in seafood (33). 2. and inadequate cooking or holding within the temperature growth range (27). when introduced or inoculated into sterile components of chilled prepared recipe dishes.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). Sources of contamination are many because of the widespread distribution of this organism in the environment. Shigella spp. L.
and are a recognised means of providing data on manufacturing standards. the Institute of Food Science and Technology (IFST) (39) or the British Retail Consortium (BRC) (40). 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. but these may be eliminated by a subsequent heat-treatment stage. For instance. cereus emetic toxin are often confused owing to similar symptoms. the fresh product is often consumed before the results are obtained.III based on IFST Guidelines (39)) provide example guidelines for seafood products. although the only seafood products specifically identified are raw “shelled and shucked products of cooked crustaceans and molluscan shellfish”. (36). 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. although due to long analysis times required. Microbial guidelines. . In the EU these are defined under European Commission (EC) Regulation 2073/2005 (38). The following tables (Table 2.A dairy-based sauce that includes a non-pasteurised cheese will contribute significant aerobic total counts. such as batter mixes.7 Published Microbiological Criteria Due to the complexity of many processed food products. 2. it is not surprising that Microbiological Standards (defined as mandatory compliance required (37)) are limited to those raw materials in which specific hazards occur. unless the criteria is for a pathogenic organisms where only non detection is acceptable. and as such the monitoring is for verification purposes rather than control.II and 2. pasta. with respect to the number and types of ingredients used. but provide the industry with an indication of the microbial level at which action is required to ensure product safety. aureus enterotoxins and B. Microbiological specifications do not necessarily relate to product safety. etc. 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. Microbial criteria and the monitoring of microbial levels within a product are important for monitoring trends within production systems. 2.6. pastry. as published by for example. are not mandatory.CHILLED AND FROZEN PREPARED FISH PRODUCTS Illnesses caused by Staph. and not to final products. It should be remembered that 47 . and their association with similar materials.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.
g.FISH AND SEAFOOD the microbial criteria for other products (rice. 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.III IFST Guidelines for processed foods . cereus Staph. aureus V. Maximum= Levels at end of stated shelf life 48 . pasta. parahaemolyticus (warm-water fish) Histamine (scrombroid fish) Indicators and spoilage organisms Organism APC (heat treated) Enterobacteriaceae E. TABLE 2. ready meals Storage: Frozen/chilled Use: Re-heated prior to consumption Pathogens and toxins Organism Salmonella spp. batter etc) should also be consulted when establishing the specifications for raw materials and final product. 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. coated fish.II IFST Guidelines for part-cooked foods – e. fish fingers. monocytogenes B. L. coli GMP* <102 Maximum 104 TABLE 2.fish products.
1. References US Food and Drug Administration. Journal of Food Science. 2005. The bacteriology of fresh and spoiling fish and some related chemical changes. Yumoto I. 6. Importance of Photobacterium phosphoreum in relation to spoilage. Aureli P.8 1.G. Ishizaki K.. Taylor S.T. International outbreak of staphylococcal food poisoning caused by contaminated lasagne.R. Recent Advances in Food Science.R. Huss H. Masuoka Y.C.J.. 38 (11)... New York. 1976. Powers E. Bartram P. Wienke A.L. 10. 12. Fish: microbiological spoilage and safety. 70-2. 215..E. Ohgiya S.. 2. Animals and Spoilage. Matsuyama H. 663-87. 1991. Incidence and levels of Bacillus cereus in processed spices. 67-73.R. Journal of Food Protection. Collette R. 167-93. Mejholm O. Gilbert R. 49 4.. 59 (2).L. Lawyer R. Journal of Milk Food Technology. 39 (10). Kumeta H.. Powers E. 166-73.K. 1962. . Van Nostrand Reinhold. England and Wales Quarterly Report. 1997. Beuchat L.. International Commission on Microbiological Specifications for Foods. Bartlett C. 96. 233-7. Martin R.R. Behling A. Shewan J. 668 -70. Pvt. Woolaway M.M. 14. 7 (24). Taylor and Francis. 1986.H. Dalgaard P..C.. Microbiology of processed spices.. 1997. in Fish Inspection Quality Control and HACCP: A Global Focus. Outbreaks of foodborne illness in humans. 5 (3). 1993.W.R. Tu A. Fish and Fisheries Products: Hazards and Controls Guidance. Eds. 1997.J. Latt T. 1991. 1311-7.. 25 (1). Slavin J. IBH Publishing Co. 13.. 11. Hackney C. 2. 47.CHILLED AND FROZEN PREPARED FISH PRODUCTS An authoritative survey of specifications can be found in Shapton & Shapton (41). Washington D.C. Cambridge. Microorganisms in Foods 6: Microbial Ecology of Food Commodities. 7.M... 9. Isolation of a Pseudomonas species from fish intestine that produces a protease active at low temperature. Proper processing and handling of raw tuna. Ltd. 1982. Microbiology of Marine Food Products. London. Communicable Disease Report. Kluwer Academic. 1976. 3. Bacterial histamine production as a function of temperature and time of incubation. 8. Pathogenic microorganisms associated with fresh produce.. 204-16.A. 373-8..M. Hobbs G. Journal of Hygiene. 1997. Communicable Disease Report. Christiansen T. Oxford.. Hoshino T. Ward D. Food Science and Technology Today.. Toxin Related Diseases: Poisons Originating from Plants. Letters in Applied Microbiology.L. 2001. 15. Food and Drug Administration. Journal of Milk Food Technology. London. Letters in Applied Microbiology. 24 (5). Brown T... Murrel H. Sakamoto T. 5.
FISH AND SEAFOOD 16.A. 23. 1987. 22. Halligan A. Springer. Jemmi T. 333-6. Gaithersburg. Eds.J. 1995. 1992. Microbiology in fishery science. Blackwell Scientific Publications. 24. Liston J. 1996.C. 472-506. Huss H. Barnes E. U.. PHLS Microbiology Digest. Blackie Academic Press. Fishing News Books Ltd. 351-72. 18. coli (VTEC) . Leatherhead Food Publications.. Brooks J.. cereus and C. International Journal of Food Microbiology. 1986..V..A Literature Survey. Granum P. Leatherhead. 1992.. SAB Symposium Series No. perfringens? International Journal of Food Microbiology. 655-60. Keusch A. Baird-Parker T.L. Clostridium perfringens in the environment.Yersinia. Microbiological guidelines for some ready-to-eat foods sampled at the point of sale: an expert opinion from the Public Health Laboratory Service. Loessner M.. Curran D. Golden D. Food preservation by combined methods. Walker S. Report on Vacuum Packing and Associated Processes. 19. Jay M. 1996. Detection of specific spoilage bacteria from fish stored at low (0 ºC) and high (20 ºC) temperatures. 1977. 1990. 1988. Ronner U.J. 59-72. 30. Trolle G. Food Taints and Off-flavours. 2000. 28.E. 26. 1979. Is Aeromonas spp. in Anaerobic Bacteria in Habitats Other than Man. in The Microbiological Safety and Quality of Food. Advisory Committee on the Microbiological Safety of Foods. Applied Microbiology. 1974. Modern Food Microbiology. Ray B. 31. Ed. Morgan D. Oxford. 20.. Food Research International. Food Microbiology. Aeromonas and Verotoxigenic E.. Connel J. Food Control. 51-158 Saxby M. Matches J. 9. 1993. Anon. 41-3.. 25 (2).M..H. Microorganisms in Foods 5: Microbiological Specifications of Food Pathogens. 27. 4 (1). Occurrence of Listeria monocytogenes in fresh water fish farms and fish smoking plants. Aspen Publishers. Lund B. Lund B. a foodborne pathogen? Review of the clinical data. 13. The Emerging pathogens . Gram L. 274-86.J. 11. 309-16. Journal of Food Safety. . Glasgow. Journal of Food Protection. Anaerobes in relation to foods of plant origin. 1994. 65.R. in Advances in Fish Science and Technology..W.J. Gould G. Blackie Academic and Professional. Her Majesty’s Stationery Office.J. 4. Wood L..C. London. 2005. Mead G.M. What problems does the food industry have with spore forming pathogens B. International Commission on Microbiological Specifications for Foods.. Fresh and processed fish and shellfish . Effects of freezing and storage on microorganisms in frozen foods: a review. Andersson A. Leistner L.R.K. Gram L. 32. 13 (1). Speck M. London. 1996.C. 28.. New York. 21. 50 17. 28 (4). Huss H. 25. 29..H. 40 (5). Eds. 155-9. Survey of the incidence of Aeromonas and Yersinia species in retail foods. Liston J. 34-40.
CHILLED AND FROZEN PREPARED FISH PRODUCTS 33. Psychrotrophic Bacteria in Foods: Disease and Spoilage. 37. Microbiological Criteria.eu/LexUriServ/ LexUriServ. 50 (9).campden. Foodborne Pathogens an Illustrated Text. Evans M. 39. U. 1997. 38. Journal of Food Protection. Wolfe Publishing. London.A.L. Journal of Food Protection. Campden & Chorleywood Food Research Association.G. British Retail Consortium. 2006. 34. Principles and Practices for the Safe Processing of Foods.F. CRC Press. 1987.A. 1991. 40. McCarthy S. Shapton N. London. Phil Voysey. 1998. Guidance on the Practical Implementation of EC Regulations on Microbiological Criteria for Foodstuffs. 51 . Woodhead Publishing Ltd. Development and Use of Microbiological Criteria for Foods. Kraft A. Varnam A.pdf Commission Regulation (EC) No 2073/2005 of 15 November 2005 on Microbiological Criteria for Foodstuffs. 60 (4).. http://eur-lex. Institute of Food Science and Technology. 372-6.org/Resources/Chilled%20Food%20Association/Public%20 Resources/BRC_CFA_Micro_Criteria_Guidance_Ed_1. Chilled Food Association. Shigella as a foodborne pathogen.uk/cday/posters2008/npd6.pdf Shapton D. http://www. http://www. 1992. Gloucestershire. 788-801. Chilled Food Association. Incidence and survival of Listeria monocytogenes in ready-to-eat seafood products. 1999.chilledfood. 35..A.H. Smith J.2. Cambridge.K. 36. U.europa.K. 41.do?uri=CELEX:02005R2073-20071227:EN:NOT Institute of Food Science and Technology.co.
The term shellfish in this context is limited to oysters.7 litres of water per hour. In the process of filtering water. slugs. oysters and clams can process about 3. can hold the shells tightly shut against enemies (1). mussels. Plankton and algae form the bulk of their diet from seawater.3. and shrimp. viruses. whelks.1 Definitions General features of the mollusc The phylum Mollusca. Pelecypoda. Pacific Regional Laboratory Northwest U. Pelecypods have little need for locomotion. and 2. where it is eventually swallowed and digested. Special tracts of cilia move this mucus toward the mouth.E. snails. 4). they also trap bacteria.6 litres. Strong muscles. The phylum consists of five classes: Amphineura. completely encloses the body. and Cephalopoda. and scallops. the adductors. Bivalves feed by filtering large volumes of water across their gills to obtain oxygen and food. abalones. cockles. The class having species of economic and public health importance is the Pelecypoda.1 3. mussels. The class Pelecypoda includes all of the bivalves such as clams. clams. and tooth shells are a group of soft-bodied animals that usually secrete external protective shells. For example. cockles and scallops. chitons. Food and Drug Administration 22201 23rd Dr. chemical contaminants and other impurities on the mucus of the gills. Many microorganisms ingested by shellfish survive the digestive process (3. S.1. Other species of shellfish such as crabs. mussels. The digestion process requires less than 2 hours in actively feeding adult bivalves (2). composed of two valves hinged together dorsally. In most species. octopi. the shell. squid. MOLLUSCAN SHELLFISH Carlos Abeyta. lying well within the mantle cavity. In these forms. the foot is compressed to form a muscular spade for digging and the head is greatly reduced. respectively. O. lobster. P. clams. which includes oysters. scallops. Jr. while 53 . Scaphopoda.S. Box 3012 Bothell WA 98041-4421 United States of America 3. oysters. cockles. Gastropoda.
1. The habitat of oysters and mussels are sessile. Clams are benthic and live burrowed in the mud.I). and usually at higher concentrations than found in their habitat. dwelling on the bottom or attached to structures in the water column. Mactra sachalinensis Saxidomus giganteus Japan Pacific – North American Saxidomus nuttali Protothaca staminea or Paphia staminea 54 . Mercenaria mercenaria Venus mercenaria Mya arenaria Country Scientific nomenclature Hard clam (also used for Quahaug/quahog) Pacific . Whatever contaminants are in the water will eventually get into the shellfish. many species of bivalves are edible (Table 3. 3. but maintain contact with the water by means of a siphon tube (5).North America Saxidomus nuttali Venus mortoni Protothaca thaca Meretrix spp.FISH AND SEAFOOD commercially valuable. are not as vulnerable to contamination through pollution of their beds as the bivalves. The following are descriptions of the various bivalves and their forms of preparation for human consumption (6): TABLE 3.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.2 International commercial products Globally.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 .
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. 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 .
2. but 56 .1 Clam Clam is the common name of a large group of often edible. or siphons. cherrystone. The muscular foot is used to burrow in mud or sand. and two symmetrical shells (valves) held closed by two large muscles and joined by a dorsal hinge joint. geoduck. A mantle tissue encloses the body of the clam. hard-shell or soft-shell clam.1.FISH AND SEAFOOD Mussel Mussel Mussel Scallops Scallop Coquille St. 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. Different species are variously called quahog. and other names. extended above the surface to maintain water currents needed for respiration and feeding. Buried clams leave tubes. mostly marine bivalve molluscs. laterally compressed. littleneck. Most clams are only a few centimetres in their maximum dimensions.
57 . Dried: Hoshigai (Japan) shelled meat. or shelled meats. and no siphon. uncooked meats packed unprocessed. having rough and irregularly shaped shells that are closed by a single adductor muscle. then heat-processed. sun-dried. Oysters lack a foot or have a reduced foot. Canned: meats removed from the shell by steaming. These molluscs feed on minute plants and animals carried to them by the current. or shelled meats. oyster soup. They live free on the bottom or adhere to stones or other objects in shallow water along the seacoasts. packed in weak brine. Live: in shell Fresh: in shell or shelled meats. after precooking and brining. Dried: shelled meats. smoked meats are also canned. 3. uncooked. skewered with bamboo sticks. clam madrilène and clam liquor. Frozen: shelled meats. trimmed and packed. Soups: oyster stew. Smoked: meats.1.MOLLUSCAN SHELLFISH the giant clam of the Indian and Pacific oceans may be nearly 130 cm long and weigh 227 kg. boiled and then sun dried for 3 to 10 days before packing in boxes (Hong-Kong) Smoked: meats usually canned in edible oil. in cans with either brine or clam liquor. all in cans. Canned: clams are steamed open and the meats removed from the shells. uncooked (shucked). Fresh: in shell. the meats are smoked and packed in oil in cans or jars.2 Oyster Oysters are marine bivalve molluscs. Live: in shells. Canned soups include clam chowder. oyster bisque. Semi-preserved: meats cooked and packed in spiced vinegar.2. tidal waters or in brackish water in the mouths of rivers. Frozen: in shell. either whole or minced. hermetically sealed. washed.
2. Smoked: meats may be canned in edible oil. They have filibranch gills. 58 . Internally. Unlike other sessile bivalve molluscs. flattened ear or wing at the hinge area.2. and held apart. smouldered and afterwards dried (Japan). by interlocking tufts of hair-like cilia. In some species. sauces.1. scallops can swim or leap by a series of rapid closings and openings of the valves.FISH AND SEAFOOD 3. brine. boiled. also with other ingredients. They have a radially ribbed shell with the edge undulated and swim by opening and closing the valves. The shells are fluted and round with a broad. cooked meats packed in dry salt for transport. vinegar solution. The familiar marine mussel is asymmetrical. Salted: cooked meats bottled in brine. Bait: used live extensively for baiting fish hooks for line fishing. 3. with the posterior ends of the shells (valves) broad and rounded. Fresh: shelled meats (often only the adductor muscle and the roe are eaten. in mussel liquor. in which the individual branches.4 Scallop Scallops are marine bivalve molluscs living in shallow waters of protected bays with sandy or muddy bottoms. scallops possess a single large adductor muscle. or filaments.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.1. or packed with jelly. the shell is often characterised with a lustrous nacreous or pearly lining inside their shells. mussel in butter sauce. Canned: cooked meats. mussel paste. The common scallop is about 5 cm long. are united. Fresh: cooked meats. composed of striated muscle fibres (used for fast swimming) and unstriated muscle fibres (used to hold together the two halves of the valves).) Dried: peeled. Live: whole with shells. The adductor muscle is the only edible part of the animal. The posterior halves of the valves contain a large adductor muscle that holds the valves closed. gutted. and the anterior end smaller and pointed. Semi-preserves: cooked meats with vinegar-acidified brine and spices.
and the environment.5 Cockle A marine bivalve mollusc having a shell with convex radially ribbed valves. Salted: meats either lightly or heavily dry salted. 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. Primarily inhabits shallow waters near tidal zones where they bury themselves in the sandy or muddy bottoms. The following pathogenic microorganisms causing illness in humans are characteristically associated with molluscan shellfish: Vibrio parahaemolyticus Vibrio cholerae O1 Vibrio fluvialis Vibrio hollisae Vibrio vulnificus Vibrio cholerae non-O1 Vibrio mimicus Vibrio furnissii 59 . clams and mussels. Canned: in own juice. also bottled in brine.1. Because molluscan shellfish are filter feeders. nutrient concentration and pollutants of the water column.MOLLUSCAN SHELLFISH Frozen: shelled meats. are vectors of disease transmission (8). Live: in shell. depending on length of journey. Vinegar cured: bottled in malt vinegar after brining. salinity. Canned: in brine. Cockles range from about 1 to 15 cm in length and have two valves of equal size.2. 3. mostly in oysters. Fresh: meats removed from the shell by boiling. pH. Whatever contaminants are in the water will eventually get into the shellfish (7). 3. and butter. Raw and partially cooked shellfish. which affects the temperature. The microflora are dependent upon a number of factors such as season. they accumulate pathogenic microorganisms from polluted waters. in sauce. The initial microflora of molluscan shellfish is comprised of the natural commensal microorganisms and the microorganisms accumulated from the water during feeding.2 Initial Microflora The microflora of molluscan shellfish directly reflects the environment from which the shellfish is harvested.
2.3. 3.1 Processing description The method of processing shellstock begins with the harvester collecting shellstock by the utilisation of tongs. At harvest.000 /g. 12. mussels.. Moraxella spp. Flavobacterium spp. Micrococcus spp.1 and 3.1. Flow diagrams of the major processing steps of molluscs are presented in Figures 3.. For example. dredges. 13). from which one or both shells have been removed. the initial flora composition and concentration of bivalves are highly variable. molluscan shellfish have an Aerobic Plate Count (APC) of approximately 1000 to 100. salinity. whole or in part. and yeast cells of Rhodotorula rubra and Trichosporon spp..FISH AND SEAFOOD Aeromonas hydrophila group Staphylococcus aureus Shigella spp.. but have never been cultured (14. The largest groups found are the Pseudomonads.. and shucked shellfish that is shellfish. and cockles The processing of bivalves takes two forms..3. (9. that have never been cultured on laboratory media. shellstock (live) which is shellfish in the shell.1 Processing and its Effects on the Microflora Oysters. Pseudomonas spp. nutrients. Campylobacter coli Bacillus cereus Hepatitis non-A. Acinetobacter spp. responding to the environment in which they are harvested. In summary. Alcaligenes spp. pollution point sources and hydrological events will greatly influence the initial microflora in bivalves. rakes. Other microorganisms found in lesser numbers are the Achromobacter spp. Bacillus spp.1 Shellstock 3. 15). have been observed microscopically in the digestive tract of Eastern and Pacific oysters.. 3. There are microorganisms. Enterococci spp.1.3 3. Campylobacter jejuni Escherichia coli Hepatitis A Norwalk virus Other viral agents A variety of commensal microflora are found in molluscan shellfish. and Aeromonads.. Temperature.1.3. spirochetes of the genus Cristispira and Saprospira. Water temperatures affect the concentrations of microflora in the water. non-B Snow Mountain virus Plesiomonas shigelloides Salmonella spp. They are termed viable non-culturable microorganisms. or by 60 . 10. observed microscopically.. 11. The microflora of molluscan shellfish at harvest consists predominantly of Gramnegative rods while the Gram-positives constitute a minor portion of the flora. mechanical harvesters. clams. Vibrios.
Generic flow diagram of live shellfish from harvester to point of sale 61 .1. or from other sources approved by the State Shellfish Control Agency (SSCA). If by boat. after harvesting. or immediately placed in refrigerated trucks. Shellstock are placed in drycooler storage containers in the facility. or in bay-approved shellfish-growing waters prior to sale or processing. The shellstock is transported dry. Water used for shellstock washing is from approved growing areas.MOLLUSCAN SHELLFISH hand (Figure 3. usually the boat will be docked by the processing facility with shellstock piled on the deck of the boat or dock. by truck or boat. Fig.1). 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. Shellstock are given a wash to reasonably free them of bottom sediments and detritus as soon as practicable. shellfish can be harvested from remote areas at convenient times and held live for brief periods at locations close to the point of sale. There are three primary reasons for wet storage: first. Secondly. for transportation to the point of sale. Shellstock may be wet-stored at the processing facility in tanks of clean seawater. Thirdly. in containers from which they can be retrieved easily. usually in open-mesh containers and/or burlap sacks. wet storage may be used to increase palatability by increasing salt content of shellfish harvested from low salinity waters.3.
22). Generally. 25). Wet storage operations are highly variable and may range from temporary storage near-shore in approved areas to on-shore tanks using re-circulating. Water used for shellstock washing should be of good sanitary quality. Over a period.1. shellstock are not refrigerated on the harvest boat. Furthermore. with prolonged storage the microbiological quality of the bivalve decreases.2 ºC (45 ºF) or below (19). shellstock are placed in refrigerated trucks for transport or placed in wet storage. synthetic seawater for the purpose of de-sanding and salt uptake. vulnificus. the bacterial counts in the meats will increase. 26). The most important factor in controlling bacterial growth in shellstock is temperature. Washing shellstock helps prevent quantities of sediment and bacteria being mixed with. 23. At harvest.2 Effects of microflora during processing The initial microflora found on or in shellstock is influenced by sources of shellstock. without adequate refrigeration. Shellfish in wet storage tanks are similarly subjected to pollution if the tank water is obtained from a polluted source.3. Shellfish may contain higher levels of pathogens than are found in the water in which they grow. when it reaches the processing facility. Excessive sediment on the shells and dead shellfish may increase bacterial 62 . are prevented. involving 691 cases was attributed to oysters contaminated in a wet storage area (27). The washing of sediment and detritus soon after harvesting will prevent further contamination. Oysters will remain alive in the shell for approximately two weeks if kept at the recommended temperatures. 17). 24. however. including V.1. Klebsiella.FISH AND SEAFOOD 3. there is evidence that in the absence of adequate refrigeration. An outbreak of infectious hepatitis in Sweden in 1956. and the sanitary practices applied during harvesting and shipping. Enterobacter. handling. At this temperature the multiplication of faecal coliforms (E. There is a positive relationship between sewage-polluted shellfish and enteric disease transmitted by shellfish (8. to avoid possible contamination of the shellstock. If long delays occur before reaching the processing facility and/or refrigeration. Shellstock should be refrigerated at temperatures at or below 10 ºC (50 ºF). Food and Drug Administration (FDA) recommends that potentially hazardous food such as shellfish be held at 7. 21. and thus contaminating the shucked meat at the point of sale. and temperature storage of shellstock to point of sale. and Citrobacter species) and Vibrios. Several studies have established that pathogenic Vibrios and other bacteria may be present in marine sediments throughout the year (21. The safety of shellfish is predicated by the cleanliness of the growing area waters from which they are harvested. hydrophila (18). 16. Generally. with the exception of A. metabolic waste accumulates in the shell serving as nutrients for bacterial growth. for a considerable length of time. some pathogenic organisms will survive and multiply in shellfish (20. coli. bivalves tightly close their shells thus trapping water and microflora. Several studies have established the increase of bacterial counts in shellstock during post harvest period before the shucking process (18. Harvest conditions are another factor that influences the growth of bacteria in shellstock.
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. through which the washed meats are discharged for rinsing and then packed in containers. shellstock is subjected to contamination when stored on the floor in standing water. the meats are in a tank of water and agitated by the introduction of air from the bottom of the tank. After washing of meats.2 Effects of microflora during processing The same considerations as previously discussed.1.1.3. If shellfish are not reasonably clean at the time of processing.2. To remove the shellfish attached to the remaining valve.1. Studies have shown the use of rubber gloves reduces the contamination level. thus contributing to high bacterial counts in the final product (28). the finished product may become contaminated. Under these conditions if the shuckers enter the packing room. and packing shellfish into containers (Figure 3. 3. for the effects of microflora during the processing of shellstock. are applied to the shucking operation of shellfish. lobsters.2). the tank is opened through a drain gate and chute. 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).1 Processing description The processing of shucked shellfish is limited to washing of shellstock. Shuckers routinely use either cotton or rubber gloves during the shucking operation. and fish. Meats are collected in either perforated or non-perforated buckets for delivery to the washing station. washing.3. whereas cotton become wet and 63 . or subjected to splash from foot traffic. Another potential risk with the use of wet storage is the commingling of bivalve molluscs with other marine species such as crab. Containers are placed in packing boxes for shipping at refrigerated temperatures. Shucking and washing of meats under sanitary conditions prevents the proliferation of bacterial populations. Washing of meats with potable water is accomplished by placing the meats on a skimmer table (the stand-supported.2. the knife is used to cut the remaining adductor thus freeing the shellfish (meats). Sealed containers are placed in boxes and covered with crushed ice for chilling or refrigeration and transportation. Thus. 3. In addition. a considerable quantity of the adhering material will be mixed into the shellfish during the shucking process. washing and culling shellfish prior to storage is essential. In the blower-washer. removal of shellfish (shucking). clothing becomes very soiled.3. The process of shucking involves forcing a knife between the valves and cutting the adductor muscle of one valve freeing the shellfish from one valve. Due to the nature of the shucking operation.2 Shucked shellfish 3.
3. Shucking buckets and storage containers are kept at adequate height above the floor to prevent contamination from floor splash. aprons. 2. There is a tendency to store such articles on the shucking benches or in packing rooms. Shucking containers are rinsed clean with running water. Shucking buckets are completely emptied at the packing room and no overage is returned to the shucker. and sanitised before each filling. Separate rooms or lockers for storage of clothing. which may lead to contamination of product. and gloves help prevent the contamination of shellfish. within one hour. Shucked meats are thoroughly drained.2.2 ºC or less. 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 . Fig 3. cleaned and delivered to the packing room or prechilled and placed in temporary refrigeration (≤ 2 hours) at 7.FISH AND SEAFOOD retains bacteria during the shucking thus increasing the bacterial load in shellfish (29). 4.
blowers and other utensils that have cracked. is stored in the bivalve as a source of energy. 34. Studies have shown that bacterial growth is significantly reduced at storage temperatures of less than 7. Temperature abuse will result in high bacterial counts. and yeasts. The high incidence of proteolytic bacteria and other types capable of fermenting glucose in the natural flora may be responsible in postmortem spoilage of shellfish. thus producing acid and gas (38). skimmers. in oysters. are an excellent medium for growth of bacteria. All equipment coming in contact with the shellfish such as colanders. The environment has an effect on the bacterial population found in shellfish. 35. Pseudomonas. these include lactobacilli. Alcaligenes. This process changes the bacterial profile to microorganisms favouring low pH. 32). Much of the work has been done with oysters and clams. rough.2 ºC or less. shucking pails. or inaccessible surfaces are apt to harbour accumulations of organic material in which bacteria may grow. Achromobacter and Flavobacterium levels decrease during storage. Shucked shellfish are held and transported at temperatures of 7. Flavobacterium. keeping shellfish refrigerated is the most effective method of retarding deterioration and spoilage. Glycogen. Shellfish.8% (37). Lactobacilli seem to be the predominant microflora in shellfish. whereas bacterium such as Pseudomonas. and Pseudomonas fluorescens are the responsible agents of spoilage in oysters held at various temperatures (10. particularly shucked meats. particularly oysters stored at 7 ºC. 3. at the turn of the century. In summary. and packing the shellfish in a timely manner.4 Spoilage There is a lack of information in the literature regarding spoilage microorganisms in shellfish. The spoilage process in shellfish may be divided into three stages: increase of acidity. a carbohydrate.2 ºC. Micrococcus. clean gloves. Refrigeration and shipping of shucked shellfish in wet ice is highly recommended. 65 . 31.47 to 6. Proteus. Earlier studies. spoilage will occur.MOLLUSCAN SHELLFISH Unacceptable practices in the packing step can affect the microflora of shellfish. Controls that should be applied include using clean packing equipment. abundant gas production. Shucked shellfish are an excellent medium for the growth of bacteria. where there is a breakdown of temperature control or prolonged storage of shellfish. As described previously. 36). streptococci. anywhere during the process of shellfish from harvesting to the point of sale. glycogen levels range from 0. and that storage in wet ice is the most effective method for refrigeration of shucked meats (30. and proteolysis (10). 33. Glycogen is hydrolysed by the initial flora found in shellfish. The initial flora found in shellfish is the major factor of spoilage in bivalves. These organisms may play a significant role in the spoilage processes. showed that organisms such as Achromobacter. For example.
Large numbers of bacteria must be present to reach potentially infective levels of >105 colony forming units per g (39). They can be introduced to the shellfish from the environment or added during processing. widespread in both polluted and unpolluted waters.5 cases/100. This bacterium is present in all the major processing steps of a shellfish plant.e. prevention of cross-contamination. V. from harvesting to finished product (i. 3. V. 66 . headache and fever. Between May and September 1997. nausea.1. Fortunately. and to the point of sale. parahaemolyticus possibly producing illnesses (40).000 population/year (46). during processing. Major outbreaks are associated with the warmer months. of V. It is a common marine bacterium. ranging from epidemic cholera to sporadic cases of diarrhoea. parahaemolyticus causes gastroenteritis lasting 24 . diarrhoea. 3. parahaemolyticus illness is usually a result of eating raw or partial cooked shellfish (8. The majority of the illnesses were associated with the consumption of raw or partially cooked oysters and clams. and estimates based from a hospital study show less than 0. shellstock and packing of shellfish in jars) (29). there is a recent report of low levels of V.5. coastal areas). Products involved were from the same processor. Diseases caused by Vibrios are enteric in nature.48 hours with abdominal pain. The ability of the bacterium to cause gastroenteritis is highly correlated with the production of a heat-stable haemolysin (42).5 Pathogens: Growth and Survival This section deals with bacterial pathogens frequently present in shellfish. and harvesting of shellfish in the cooler months.1 Vibrio parahaemolyticus V. 44). approximately 23 cfu/g. parahaemolyticus from shellfish is low. Descriptions of outbreaks or reports of growth of the organism in shellfish are presented. Sporadic cases are frequent along the coast of the United States. These organisms can be present in the shellfish from the point of harvesting. and very common in Japan. primarily in brackish or saltwater. The incidence of V.FISH AND SEAFOOD 3. lot and retail outlet. only a small percentage of strains in the marine environment are potentially pathogenic (45). where outbreaks occur regularly. parahaemolyticus. however. The control of growth and survival of these organisms can be reduced by the use of good manufacturing practices including immediate and proper refrigeration. One of 250 cases reported involved shucked products with low levels. 41).1 Vibrios Vibrios are Gram-negative facultative rods naturally occurring in the marine water (estuaries. although non-hemolytic strains have been associated with illness (43. parahaemolyticus multiplies rapidly at temperatures above 20 ºC.5. there were over 250 cases of illness on the West Coast of North American extending from California to Canada.
It causes diarrhoea. 46. Oysters are the most hazardous food associated with V. or immunosuppression.MOLLUSCAN SHELLFISH Control of V. Healthy individuals are susceptible to gastroenteritis. like V. vulnificus are fever.1.II (8. Primarily. 49). Hazards from this bacterium can be controlled by thorough cooking of shellfish. and rapidly become inactive at low storage temperatures. vulnificus. however. cholerae non-01 has been associated with septicaemia. nausea. The widespread distribution of this bacterium is comparable to other studies in the Gulf and Atlantic Coasts (52. Death has been reported to occur within 36 hours from onset. gastroenteritis. V. chills. 48). Inadequate refrigeration will accelerate the growth of V. the organism has also been found in cooler temperatures from estuaries of Washington and Oregon (47). The occurrence of V. 56). The organism attaches to the intestine and release of a toxin is suspected. cholerae.8 case/100. 3. They are sensitive to cold. cholerae non-01 is a common bacterial cause of molluscan shellfishassociated illness. diabetes. and preventing cross contamination once the product is cooked.1. parahaemolyticus. High-risk individuals. 58). The bacterium is widespread in estuarine waters particularly in warm waters of >20 °C. V. parahaemolyticus the estimated annual incidence rate is very low. these individuals are advised not to eat raw shellfish. cholerae non-01 is ubiquitous in the estuarine environment. 54. Again. The bacterium is an estuarine species that causes wound infections.0. or primary septicaemia.2 Vibrio vulnificus The symptoms of V. cirrhosis. 57. lasting 6 .3 Vibrio cholerae non-01 Illness caused by V. 55.48 hours of onset. abdominal cramps. and vomiting. Control of V.5. V.5. are particularly susceptible to primary septicaemia. vulnificus causing septicaemia and death has been associated with the consumption of oysters (8.4 .000 population/year (8. leukaemia. those who have liver disease. Onset of symptoms occurs in 48 hours and can be severe. and nausea within 24 . vulnificus is by rapid refrigeration during warm weather months. the organism has been found in the Gulf of Mexico. parahaemolyticus growth in shellfish meats is temperature dependent. with a fatality rate of 50% for individuals with septicaemia. 53. 46. which have been implicated in shellfish-related disease outbreaks (31). fever. in the range of 0. primarily in immunocompromised individuals with a mortality rate exceeding 50% (50). cholerae non-01 is generally a less severe gastroenteritis than that caused by V. 23 of 24 major USA-West Coast estuaries were positive for V. cholerae non-01 (51). All isolates are considered pathogenic. Control of oyster-associated 67 .7 days. Illnesses are usually associated with the consumption of raw oysters as described in Table 3. It is one of the most severe foodborne infectious diseases. 3. In one study.
vomiting. V. In developing countries.S.5%) and 2 .10 days. and shock. cholerae 01 in molluscan shellfish. Onset of disease is 6 hours to 5 days. Illness can include abdominal cramps. the pathogenic severity is lower than V. Poor sanitation and contaminated water supplies will spread the disease.5 Other Vibrios spp.10% carbon dioxide for optimal growth conditions. 3. nausea. Gulf coast environment. 68 . There is evidence of strains of V. jejuni requires reduced levels of oxygen (3 . Death may occur after severe fluid and electrolyte loss. dehydration. It causes diarrhoea. They are common marine bacteria. particularly in oysters. Serotype Ogawa biotype El Tor and 0139 are responsible for epidemics in South American. salinity. 62.1. The frequency of infection of these organisms is no different from other Vibrios. cholerae 01 is transmitted by faecal contamination of food or water.FISH AND SEAFOOD V. or cross-contaminated shellfish (57. imported product from a country experiencing an epidemic. In general. 3. associated with shellfish-borne illness outbreaks. V. nausea. headache and fever. 60).5. cholerae 01 establishing in the U. The control of V. undercooked. fever. hollisae. preventing crosscontamination of cooked seafood. widespread in both polluted and unpolluted waters. mimicus. acidity and heat. is the same as previously discussed for the non-01 strains. and V. cholerae non-01 illnesses is by cooking seafood thoroughly.48 hours with abdominal pain. and illness lasts 7 .2 Campylobacter jejuni C. whereas excellent sanitation facilities in developed countries are responsible for the near eradication of epidemic cholera. Onset of illness is from 3 to 5 days after ingestion of contaminated food. V. (8. The presence of these strains appears to be in low numbers and non-virulent (59). 63). drying. 3. these organisms cause gastroenteritis lasting 24 . Control of infection from these organisms is no different from the other Vibrios.5. including V.4 Vibrio cholerae 01 and 0139 Symptoms of epidemic cholera vary from mild watery diarrhoea to acute diarrhoea with characteristic rice water stools.g. cholerae 01. It is considered a fragile and sensitive organism to environmental stresses such as oxygen. and nausea. low pH. however.1. abdominal pain. Illnesses are associated with the warmer months. fluvialis. 61. vulnificus. In addition. and harvesting of oysters in colder months when Vibrio counts in water are the lowest (59). processors should know the product source. respectively. and southern Asia including India and Bangladesh. There are several other Vibrio spp. diarrhoea. parahaemolyticus and V. and the transmission of the disease by consumption of raw.5. e.
76. and have been implicated as the causative agent in illnesses from the consumption of raw oysters and clams (68. Sixteen of 28 participants became ill with campylobacteriosis after ingesting raw clams. they survive well at refrigerated temperatures. 81. shigelloides (79. from Pacific shellstock oysters purchased from a retail grocery store (74). the organism is being continually inoculated into the environment by waterfowl and farm run-offs and thus eventually contaminating shellfish growing waters. 85). and their immediate proper refrigeration. Recently. Campylobacters do not survive well in the environment due to the organism's lack of resistance to environmental factors. have been isolated from shellfish beds throughout the world (67. 71).3. Campylobacter spp. 78) and P. 65. V. Human feeding studies suggest that about 400 . whereas at 10 ºC Campylobacter survive only for 5 days (71). mimicus.4 to 114 cells. control of C. fluviallis. 66). hollisae Infective dose varies with host susceptibility and virulence of the strain. jejuni does not multiply in shellstock at refrigerated temperatures. 73). 82.II Molluscan shellfish associated outbreaks caused by Vibrio spp. although their role as a pathogen is highly controversial and circumstantial. and V. 69.5. This finding is significant in that C.3 Other organisms of concern 3. V. vulnificus. the incidence of Campylobacter infection is low from the consumption of molluscan shellfish. 3. Proper care should be taken if the finished product is to be packaged in modified-atmosphere packaging (MAP). V. C. jejuni survived for over 20 days in market-shucked oysters stored at 4 ºC. 77. cholerae 01 and non-01. Although. 70. Campylobacter appear to survive well in oysters during storage. infection occurred among participants of a firemen's banquet. C. The seasonality of 69 . They have been implicated in limited shellfishassociated gastroenteritis cases (8. hydrophila group (75. jejuni were recovered. of Cases 4 362 0 1 0 a Includes V.a (8) Species Hard clams Oysters Soft Clams Mussels Scallops No. 84. 72.5. ranging from 0. To reduce the risk of shellfish-borne campylobacteriosis. however. V. parahaemolyticus.1 Aeromonas hydrophila group and Plesiomonas shigelloides A. however. In one case.500 cells/gram of Campylobacter may cause illness (64. jejuni begins in the harvest of shellstock from approved waters. 80) are ubiquitous in the marine environment. of Incidents 4 279 0 1 0 No. 83. 68.MOLLUSCAN SHELLFISH TABLE 3.
being isolated more often during the warmer month. non-dairy frozen desserts.000/gram (29). FDA/Administrative Guidelines 3 Source (91). aureus C. Caution should be exercised by individuals with haematological malignancies (88). action is considered on a case-by-case basis taking into account all the factors associated with the specific situation. Control of these organisms in shellfish products is limited. cholerae V. jejuni V. 87) of the outgrowth of A. parahaemolyticus Staph. vulnificus in molluscan shellfish. 4 FDA and ISSC recommending establishment of an educational program to alert at risk population of presence of V. They survive well in shellstock or shucked oyster meats held at refrigeration temperatures. coli Enterotoxigenic E. FDA established action levels or levels of concern for certain microbial pathogens in molluscan shellfish. Pathogen monitoring of Selected High Risk Foods Domestic including: milk. dried milk/whey drink powders. and where indicated are intended to be used for official control purposes. soy products. suggesting that these individuals are at higher risk when consuming raw or partially cooked shellfish containing these bacteria.FISH AND SEAFOOD Aeromonas and Plesiomonas is similar to the Vibrios. They are easily destroyed with mild cooking temperatures.000 cfu/g1 <230/100 g2 <1000 per gram. hydrophila group stored at refrigeration temperatures.600.000 to 3.000 MPN per gram1 negative for staphylococcal enterotoxin or when the viable MPN count is <10. 2 Source (90). 3. In one study.0001 negative for the presence3 Under review4 Source (89). Test Standard Plate Counts E. Similar results were seen by other investigators (86. after 7 days storage of shucked oysters at a shellfish processors’ cooler kept a 5 ºC Aeromonas counts ranged from 500. coli Salmonella V. Note: Although molluscan shellfish are not included in the selected high risk food.6 Published Microbiological Criteria The published microbiological criteria below are included as a guide. Enforcement action is considered on a case-by-case basis taking into account all the factors associated with the specific situation. prepared sandwiches. vulnificus 1 Acceptable Levels in Molluscan Shellfish <500. 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. The Agency will consider enforcement action against the shipment of molluscan shellfish if the levels exceed as stated. FDA/ Administrative Guidelines. 70 .
. 4. Journal of Milk and Food Technology.K. Smith F. 14. 3. Bacterial flora of the market oyster. 29. 13.. 1982. 1984. 82-7. Fleet G.7 1. American Journal of Hygiene. 26.S. Microbiological characteristics of Dungeness crab (Cancer magister). 6. Wiley-Blackwell. Philadelphia.F. water and sediment from Galveston Bay.A. 245-7. 1983. Rippey S. 6. Tall B. 105-8.W. 33643. Microbial flora and level of Vibrio parahaemolyticus of oysters (Crassostrea virginica). 1973. 16... in Fishery Bulletin 1964 No. 13. Microbiology of bivalve molluscan shellfish. Eliot C. 1960. Viability and release Salmonella charity and Escherichia coli from oyster faeces. Applied and Environmental Microbiology. 1981. U.W.D. 755-76. Schmidt C. Tubiash H.. Microbiology of shellfish: bacteriological study of the natural flora of pacific oysters (Crassostrea gigas).A.R. Colwell R. 64. Eds. 12.R. Gill S..MOLLUSCAN SHELLFISH 3. Galtsoff P.J. 11. W. Quantitative and qualitative commensal bacterial flora of Crassostrea virginica in Chesapeake Bay.. 9. 1975. References The Phylum Mollusca. New York. Effects of harvesting waters and storage conditions on yeast population in shellfish. 419-25. 1926.K. 1963. Organisation for Economic Co-operation and Development. Lovelace T. Jr. 544-8.R. Van Nostrand Reinhold. 10. 7. 1940. Lee J. Ray S.R.C. 2. 30. Bernard F. Thompson C. Proceedings of the National Shellfisheries Association. USA. Cook D. 8. Jr.. 33-6. 1968. Nauman R. Villee C.R.N. American Journal of Public Health. Rowse A. Ward D. Clinical Microbiology Review. 42.E. 30..S. 1991. Scanning electron microscopy of Cristispira species in Chesapeake Bay oysters. 46.M. in Microbiology of Marine Food Products. Fish and Wildlife Service. Hood M. 44.R. Infectious diseases associated with molluscan shellfish consumption. Fate of enteric bacteria in estuarine sediments and oyster faeces. 15.E. Galtsoff P. Saunders Company.S. The American Oyster. 7. Ed. 19-34.L. 447-52. 17. Veliger. 8. 71 . Cook D.. 633-40. in Multilingual Dictionary of Fish and Fish Products. Applied Microbiology. 1964. 1970. 71-6. 58.A. 231-51. 1936. Colwell R.. Liston J.H. 104-9. 1995. Washington D. 36. Walker W. Applied Microbiology. Eds. Government Printing Office.S. Biology of the oyster in relation to sanitation. Hackney C.. in General Zoology. American Journal of Public Health. 72-8. Occurrence of the spirochaete genus Cristispira in western Canadian marine bivalves. 5. Applied and Environmental Microbiology. Old H. Journal of Food Protection. Vanderzant C. 1994. Pfeifer D.. A typhoid fever epidemic caused by carrier bootlegging oysters. Journal of the Mississippi Academy of Sciences.B.C.
Tokyo. Maryland.. Eds. Blake N.A. Special Report U. Hoff J.A.. Ed. Public Health Report. Public Health Service. U.S. 19. 20. Presnell M. 47 (5). Kaysner C. 40.J. Part II Harvesting.. 167-71. Kelly C. 1983.J. Food and Drug Administration..B. Jones W. 343-9. 53.C. Ericksen T. Vasconcelos G. Ness G. Vaughn M. 1973..M. Behaviour of pathogenic bacteria in the oyster.. Wekell M. Fujino T. Liston J. 1989.J. 22.. 324. 24. 25. Chesapeake Science. Presnell M. 30. Timetemperature effects on the bacteriological quality of stored shellfish. 27. Handling and Shipping Shellfish. 1214. Influence of U.H.F.S.. Bacteriological survey of an oyster bed in Tangier Sound. 1964. 123-8. 45. Journal of Food Protection.. 1980. Applied and Environmental Microbiology. Crassostrea virginica and Mercenaria campechiensis. 28. Abeyta C. 31. 29. 1954. I. Bacteriological studies of commercial shellfish operations on the Gulf Coast. Bacteriological changes in live shellfish: Pacific oysters (Crassostrea gigas). 1971. 1967. Colburn K. Fleet G. FDA Gulf Coast Technical Services Unit. 1205-10. and Manillal clams (Venerupis japonica). Journal of Food Protection. Beck W.H. Establishment of critical control points in west coast oyster processing plants.W. 1961. The influence of time and temperature on the bacteriological quality of shell oysters during processing and shipping. Koburger J.. 1995. 1984. Rodrick G. seafood-handling procedures on Vibrio parahaemolyticus. Incidence of Samonellae in clams.FISH AND SEAFOOD 18. P-1.. 1992. Ltd.A.. 69. Williams L. Indicator bacteria and Vibrionaceae multiplication in postharvest shellstock oysters. Kelly C. Arcisz W. Sakazaki R. Cook D. Crassostrea commercialis.G. Svenska Lakartodningen. U. 989-1003.W.. Applied and Environmental Microbiology. oysters. LaRock P.B. Trost P. and storage. Olympia oysters (Ostrea lurida). Journal of Food Science. Washington D. U. in National Shellfish Sanitation Program Manual of Operations. during depuration. Native Littleneck clams (Protothaca staminea)..D. Stott R. 23. Applied and Environmental Microbiology. Takeda Y. 1221-8. F61-9. Food and Drug Administration.W. Fraiser M. in International Symposium Vibrio parahaemolyticus. Survival of enteric organisms in shellfish. 4..S. American Society for Microbiology.T. 72 ..S. 5. Hood M.C. 1490-5. Food and Drug Administration. Effects of storage on microbial loads of two commercially important shellfish species. re-laying.A. Son N. 26.E. crabs and mullet.A..E.W.. 343-5. Ruple A. 32. 1956. Jr.S. Hepatitis epidemic conveyed by oysters.A. 21. Saikon Publishing Co..W. Temporal occurrence of Vibrio species and Aeromonas hydrophila in estuarine sediments. 1985. 50.. 994-1002. 52.B. Roos B.. Sadaguchi G.. Public Health Service Technical Report.
A. 69... 1916..C. Bacteriological examination of oysters stored at low temperatures.. 204-5.. Jacobson M. Volume 1. 36..W.T. Applied Environmental Microbiology. Powers C. DePaola A.A. Libonati J. Crassostrea virginica. 1989. Serological and hemolytic characteristics of Vibrio parahaemolyticus from marine sources.... Takikawa I. Novak A. 39. Borgstrom G. Kelly M. Joseph M. 33. Geiger J.G. Miyamoto Y. West Coast States and British Columbia. 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. 53. 46. Infectionskrankheiten und Hygien. Abeyta C. Academic Press. Sidwell V.. Thompson C. New York. 2. 353-7. Yokohama Medical Bulletin. Hlady W. U. 45. 40.M. 41. 1.. 13-7. Sample T. Journal of Infectious Disease. 44.A. Takizawa K.. Abbott S. Vibrio parahaemolyticus illnesses. Parasitenkunde. 37. Food and Drug Administration.. 1987. Watsky D. Kato T.S. 1914. Fieger E. Studies on pathogenic halophilic bacteria. 42. 313-22. 1989. Israel E.F. Public Health Reports. 38. 273-80. Vibrio infections associated with raw oyster consumption in Florida. 1954. (2). kanagawa-negative Vibrio parahaemolyticus from patients and the environment in the Pacific Northwest. Ed. Akiyama S. Marine Fisheries Review.D. Jr..N. Leitch J. 97. Journal of Clinical Microbiology. U.. 1969. 1147-9. 128-31.W.. 1997.. Bothell. Journal of Infectious Disease. Yamai S. Zentralblatt für Bakteriologie.MOLLUSCAN SHELLFISH 32. 1349-51. Vanderzant C.. Kelly C. 73 .E... 34. Urease-positive. 2820-2.. Takeda Y. 35. 1976. 160.L. 1937.B. The bacterial content of market oysters.L. Obara Y. Berry F...M..M. Wekell M.. Stott R.. Grodner R. 43.S. 133-47. 2891-3.M. Shellfish Specialist.. Geographic and monthly variation in composition of oysters. The bacterial flora of market oysters. in Fish as Food.. 1961. 41. Janda J. 1926. Virulent strains of Vibrio vulnificus isolated from estuaries of the United States west coast.. Stroh E. Journal of Food Protection. 1997. Joseph S. 25. Bacteriological control of oysters during processing and marketing. Loomis A. Hoge C. Journal of Bacteriology. Kaysner C.. Peeler R. 27. Tanikawa E.M. Ward W. 1979. 1958.P. Epidemiology and spectrum of Vibrio infections in a Chesapeake Bay community.D. 27. 41. Journal of Bacteriology. Morris J. 100. Journal of Food Science. Bacteriological findings in Baltimore oysters. Kaysner C. 38. 107-8. Ishibashi M. Arcisz W.A. 60. Jr. In vitro haemolytic characteristics of Vibrio parahaemolyticus: Its close correlation with human pathogenicity. 47. Washington Report October 14. 1989.G. Bulletin of the Johns Hopkins Hospital. 985-93. Journal of Clinical Microbiology. Microbiology of shellfish decomposition.
Blake P. Lockman H. McFarland L. Allegra D. An epidemiological study of V. 1982.E.G. 52. Wachsmuth I.. 8. Vibrios in the Louisiana Gulf Coast environment..R. Twedt R..G... Journal of Infectious Disease. McFarland L. Tacket C.C. serology.M. Jr. Jr.. 1475-8..J. Non-O group 1 Vibrio cholerae gastroenteritis in the United States: Clinical. Ed. Lee J. 802-6.. Lancet. Roberts N. Hug A. 94. Illness caused by Vibrio damsela and Vibrio hollisae. Johnston J. Safrin S. Plaard R.. 1980. Characterisation of Vibrios cholerae isolated from oysters. Davis B. Craig J.W.O... 1983....I.. Journal of Food Protection. Review of Infectious Disease. 59. Motes M. 49. Rahman R... Stott R.. 1344-8. Cholera . McCay C.M. Clinical features and an epidemiological study of Vibrio vulnificus infections.W.. Morris J... Hug M. Zywno S. Roderick G..W.A. 1. 2050-3... 74 . Becker S.D. Applied and Environmental Microbiology.G.T. Duran W. Blake P. sediment and waters of the U.. Wathen H. in Vibrios in the Environment.M. cholerae in Louisiana. 51. Jacobs R. 1981.G. Riddle C. Jr. 1984. 1012-7. Wekell M. 53.M.. Colwell R. Microbial Ecology.. 275-83. New England Journal of Medicine. 58. Leitch J.. 305.. Conte J. Applied and Environmental Microbiology. 46..T. West P.A.. Ecology. 656-8. Colwell R..T.G. Barrett T... Colwell R. Spite G. 312.M.F. 57. Joseph S. 56.W.A. Becker R. 1979.E. New York. Non-01 Vibrio cholerae bacteraemia: Case report and review.C.G..A possible endemic focus in the United States.F. Vibrio vulnificus: Man and the sea. Wilson T..M.R. Weaver R.E. and enterotoxin production of Vibrio cholerae in Chesapeake Bay.. DePaola A. DePaola A.. Pons V. Peeler J.M..M. 5972. Journal of the American Medical Association. McPhearson R. Kaysner C. 60.O..V. Gulf Coast.T. Maden J.. 45.E.N.A. 55. 1985..K.A. 253.. Caraway C.. 54. Presnell M. Blake P. Miller H... Siebeling. and laboratory characteristics of sporadic cases.O. Morris J...A.T. R. Feldman R. 1988.. Jr.B.J.J.A. Incidence of Vibrio cholerae from estuaries of the United States West coast. 1985. John Wiley & Sons Inc. Hebert S.R. Kaper J.. Morris J. Francis D.. Twedt R.. Jr. Wazenski T. Hickman F. Hunt J.F. Kaper J. 558-61.S. 305-9. Morris J. Non-01 Vibrio cholerae in shellfish. 149.M.G. 50... 41.. Tacket C.G. 299-312. Adams M.. New England Journal of Medicine. Jr. Brenner F. Bradford H. Hollis D.B. Feeley J.. Annals of Internal Medicine. Ecological relationship between Vibrio cholerae and planktonic crustacean copepods. Jr. 343-50... Black R. 1983.D.. Small P.B.L..A. epidemiologic. Cholera and other vibrioses in the United States. Puhr N.P.A. 91-103. 53. 1987. 1984. Applied and Environmental Microbiology.R.. 1981. Applied and Environmental Microbiology.. Bradford H.FISH AND SEAFOOD 48... 1294-7. Abeyta C. 1982. Wilson R. Blake P. 10. 61. Snyder J. 37..
. Fitzpatrick M.A. 69.G.M. Eds.A. 1988. Wareing D. International Journal of Food Microbiology. Wekell M. Murphy P.G. 63. Journal of Food Protection.D. 257. 607-9..A.R. 1996. Black R.. Blake P. Helicobacters and Related Organisms.. 1983. Alam A. Journal of the Medical Society of New Jersey. Ketley J.M. 1983. Ketley J. Plenum Press.. 1980. 72. EF-6. 282. Johnston J. Feldman R. Wilson I.G.G. Proudford R. The occurrence of Campylobacter jejuni and Campylobacter coli in Sidney rock oyster.R.. 56..D.G. Davis J. Davis B. Journal of Infectious Disease. London.. Skirrows M. in Campylobacter II: Proceedings of the Second International Workshop on Campylobacter Infections.A. 68. 65. Eds.M. Black R. New York.R.. 1981. C. from patients with diarrhoea. Abeyta C..A.B. Blaser M. 323-5. Huges T.A.G. Plenum Press.P. Kaysner.MOLLUSCAN SHELLFISH 62.A.M.E. 157. Disease from infection with Vibrio mimicus. Hughes T. Arumugaswamy R. 169-71.R. 73. Eds. Phenotypic and genotypic characterisation of urease-positive thermophilic Campylobacter (uptc) isolated in N. 1987. Public Health Laboratory Service. 99... Kaysner C. Robinson D. Matsuda M..E. Jr.E. 13.. British Medical Journal.. Austin S..A...G. in Campylobacters.... Brussels. Dalley E. Wilson T.. Journal of Rapid Methods and Automation in Microbiology.M.J. 621-4. 1993. New York.A. 75 . Morris G. Kaysner C. 6-9 September 1983.P.M.K.J. 67. Jones D.S. Stott R.. Huq M. 51-64. Abeyta C. Feldman R. Murphy P.. Campylobacter jejuni in a Washington State shellfish growing bed associated with illness.. Levine M.. Newell D. in Campylobacters.. 256.. 4. Blaser M.. infections in man. Journal of Clinical Microbiology. Campylobacter gastroenteritis associated with raw clams. Pearson A. Experimental Campylobacter jejuni infection in humans.S. 66. Microbial Ecology in Health and Disease.M.. Wilson I.H. Clements M. Ireland.K. (jejuni. Griffin M. coli and lari) from shellfish and the marine environment. 4. Infective dose of Campylobacter jejuni in milk. 11...M. Studies of Campylobacter jejuni infection in volunteers. Incidence and survival of thermophilic campylobacters from shellfish growing water: media evaluation. Wareing D. 1991.K. Hunt J. Helicobacters and Related Organisms. Deeter F.. 71. Abeyta C.E. Wilson T. Levine M. Wekell M. Clements M. 64...I....F. 1995.M.. A method for isolation of Campylobacter spp. Brenner D.. a newly recognised Vibrio species. 80.R.W.L. 1980. 4. 70. Moore J. 1584.J. Part 1 Abstracts: Shellfish as a possible source of thermophilic Campylobacter spp.L. Shandera W. 472-9. Jr. Rowe B.J. 1996. Jr... Newell D. Annals of Internal Medicine. Moore J.X. 101-4. Isolation of vibrio-like group. S41.
. Journal of Food Protection.L. Baker R. Burke V. Hunt J.A. Wekell M.H. 223-47..C. Annals of Internal Medicine.. Kaper J.M.. Oyster borne disease outbreaks in St. 1989. 76 . and treatment of Plesiomonas shigelloides diarrhoea. 77. 52.. Wekell M. 80. Recovery of Aeromonas hydrophila from oysters implicated in an outbreak of foodborne illness.N. 1981. Fliermans C. Journal of Food Protection. Infection and Immunity... III. 1987. 1989. Stelma G. 1985.M. Herrington D. Indicator bacteria and Vibrionaceae multiplication in postharvest shellstock oysters. Atlanta.W. Colwell R.J. Herrington T. Journal of Applied Bacteriology. Biotyping and virulence factors in clinical and environmental isolates of Aeromonas species. Report (1/13/84)..A. Sullivan J. Peterson D. 76. Koburger J. In vitro and in vivo pathogenicity of Plesiomonas shigelloides. 81. Tzipori S. Miller M. 998-1001... 85.L.A... 598-601. Applied and Environmental Microbiology. Stott R. 53..P. Applied and Environmental Microbiology. GA. 1984..R. Lockman H... Hirsch R.. Holmerg S.A. Farmer J.. Robinson J. 39. Journal of Clinical Microbiology.. Kaysner C. 449-57. 343-9. Beaman J. 1988.M. Journal of Food Protection..L.. 1997. Levine M. 55..M. 50.FISH AND SEAFOOD 74..J. Jr. 36.. 1990.. Bark D. 47. 731-8.W. Wachsmuth I.. 82. Plesiomonas enteric infections in the United States. Partridge K. Trost P. Tenge B. The use of bacterial membrane fractions for the detection of Campylobacter species in shellfish. Effects of processing and storing oyster meats on concentrations of indicator bacteria. Plesiomonas shigelloides: An opportunistic food and waterborne pathogen. Jr. 84. Robins-Browne R.M.A. 690-4. Frequent isolation of Edwardsiella tarda and Plesiomonas shigelloides from healthy Zairese freshwater fish: A possible source of sporadic diarrhoea in the tropics. Abeyta C. Applied and Environmental Microbiology.M.. Kaysner C.. 1146-9.A.T. Cooper M.B.D. 1978. 1980. Prevalence and distribution of Aeromonas hydrophila in the United States.W. 75. Incidence of motile aeromonads from United States west coast shellfish growing estuaries. 52. Gracey M. 359-77. 105... Journal of Rapid Methods and Automation in Microbiology.F. 48..J. Cook D. 83.M. 5... Ruple A. Journal of Food Protection. Clinical features.R. Abeyta C.D. 979-85. Hood M. Petersburg. Tall B.. Aeromonas hydrophila: ecology and toxigenicity of isolates from an estuary.. Department of Health and Human Services/Public Health Service/Food and Drug Administration. Journal of Food Protection. Singelton F. Abeyta C. Kaysner C. vibrios and Aeromonas hydrophila.. 849-55.. Hickman-Brenner F. 7-12. 27. Vandepitte J.K.. Jr.A..A. Van Damme L.J. Fla. 87.D. Wekell M. 1984. 475-9. 47. Seidler R. Blake P. 78. Kelly M. 1986. Kain K. Esch G. 79.C. Hazan T. 86.. epidemiology.
Ed. 3. U. 89. U. mussels.R.E. Food and Drug Administration. Pittsurgh.W. 91. Food and Drug Administration. Fish Inspection.S.C. Slavin J. 1995.. Food and Drug Administration Compliance Policy Guides. 7. Gould G. Program 7303.8 Further Reading Ahmed F. U.. Clams. Smith J. Ed.W.1987. in National Shellfish Sanitation Program Manual of Operations.MOLLUSCAN SHELLFISH 88. U. 1987. in U. 1995. U.C. Seafood Safety.. Inc. 560. and HACCP: A Global Focus.S. Washington D. Government Printing Office .H.S.. Ed.S. 1997.12). Martin R.S.S.C. Food and Drug Administration. Sherlock C. Washington D.C. National Academy Press. The Microbiological Safety and Quality of Food. 1991. Lancaster.E. Food and Drug Administration. U. Gaithersburg. Lund B.A. Burdge D...030.M. Quality Control.L. Technomic Publishing Co.600 (CPG 7119. Food and Drug Administration. Chapter 5: Foods. 2000. Collette R. 90.. oysters. Food and Drug Administration. Food and Drug Administration. Food and Drug Administration. Food and Drug Administration Compliance Program Guidance Manual. Baird-Parker T.S. fresh or frozenadulteration by bacteriological contamination Sec. 77 . Washington D.. in U. Aspen Publishers Inc. Part I Sanitation of Shellfish Growing Areas. Does Aeromonas hydrophila preferentially colonise the bowels of patients with hematologic malignancies? Diagnostic Microbiology and Infectious Disease.Pathogen monitoring of selected high risk foods (FY 88/89).S.. 63-8.
Crayfish..5 kilo pack (boxes or bags). FRSPH Ethical Trade and Food Management Group Natural Resources Institute University of Greenwich Central Avenue Chatham Maritime Kent ME4 4TB United Kingdom 4. Peeled and cooked shrimp are ready-to-eat after controlled thawing. and frozen in 1 and 2. the front pair ends in claws. Species eaten from tropical waters include Penaeus spp.5 and 1.1 Definitions Crustaceans are invertebrate animals and represent a class of the phylum Arthropoda.4. jointed limbs and a chitinous exoskeleton.. Such products are usually only partially cooked before freezing. and available in 0. prawns and scampi are temperate species commonly eaten by man and include members of the families Nephrops norvegicus (scampi). Pandalus montagui and Crangon crangon (inshore shrimp).Ph. peeled product. Pandalus borealis (deep-water shrimp). lobsters. and Trachypenaeus spp.0 kg polythene bags. with shells and heads on or off are usually packed. packed and frozen in 0. Shrimps. Breaded shrimp and scampi are battered and coated in breadcrumbs and sold frozen to the consumer for cooking prior to consumption. with additional water to form a glaze. (deep-water shrimp). 79 . crabs.0 kg polythene bags. lobsters and crabs have ten legs. Peeled ready-to-cook shrimp are raw.5 and 1. crayfish and shrimp (1)..g. e. M. and are usually aquatic. CRUSTACEAN SHELLFISH Linda Nicolaides. Shrimp can be purchased in the raw or cooked state in a variety of forms: Entire shrimp. They possess a segmented body. Parapeneopsis spp.
Shewanella putrefaciens and 80 . lobsters and crayfish are traditionally kept alive after capture and then cooked to kill immediately before consumption. particularly for members of the genus Vibrio (2). traditionally. the potential exists for crosscontamination from both the processor and from the environment. the flora will change depending upon the methods of handling and/or the environmental conditions to which they are exposed. Poor hygienic procedures will increase the probability of contamination with bacteria of public health significance. lobster tails and cooked crabmeat presented decoratively in their shells. the geography of the area they are captured from or are grown in. The microbial flora of crustaceans from temperate waters differs from that of crustaceans from warmer. 4. or frozen crab. for crabs and lobsters. which include the environment from which they have been captured or harvested. stored and transported as frozen product.g. so that spoilage may occur during landing and transportation to the processing plant. Control measures to prevent these potential hazards occurring include training and supervision of operatives as well as Good Hygienic Practices (GHP). particularly after it has been cooked. Pasteurised crabmeat is edible meat removed from the exoskeleton by hand. Cooked and chilled. it is important that shrimp are handled and iced correctly in order to keep them in as fresh and uncontaminated a state as possible. Following capture or harvest. only live carcasses are processed. lobster and crayfish are available for the consumer owing to the growing demand for ready-to-eat products. the season. spoilage bacteria are of little importance as. hence. boiled to kill the crustaceans and cook their meat prior to consumption. this presents more favourable conditions for growth by both spoilage and pathogenic bacteria. their feeding and living habits. tropical waters.g. Spoilage begins following death once the immuno-response system of the crustacean shellfish fails to function. In comparison. e. and the temperature and quality of the waters in which they exist. storage above the recommended storage temperature. during the peeling of cooked shrimp. Microbiological risks associated with crustacean shellfish increase in proportion to the degree of handling that the product undergoes. the level and type of the initial microflora of crustacean shellfish will reflect a combination of factors. with the increasing demand for ready-to-eat/cook crustacean products.FISH AND SEAFOOD Crabs.2 Initial Microflora As with finfish. As with finfish. shrimp die immediately after capture. For example. The structure of the circulatory system of crabs is not closed. e. packed into polythene bags. there is a growing market for fresh lobster tails or cooked crabmeat in crab shells. which means that the haemolymph can be a reservoir of bacteria. If these products are subjected to temperature abuse.. However. Spoilage bacteria associated with temperate crustaceans include Pseudomonas spp.
. or cooked. up to 16 days for tropical species compared with 8 . After capture and transport to the processing plant. so they are either sold on the local market or are processed in local processing plants. and Achromobacter spp.10 days for temperate water species (3). The latter process can expose the shrimp muscle to incidents of cross-contamination if GHP are not followed. corynebacteria. with “head on” or “head off” whereby the edible tail is removed from the head.1 Prawns and shrimps An example of the process used for preparing frozen.for example. glazed and frozen (see Figure 4. peeled and cooked. Shrimps can be processed whole. They are then taken to market alive in either holding tanks.CRUSTACEAN SHELLFISH members of the Acinetobacter-Moraxella group. lobsters and crayfish are harvested from cages or pots containing bait. the genera associated with tropical species include Enterobacteriaceae. In contrast. A large proportion of shrimps are produced by aquaculture. however. or with the shell and central vein removed. 4. uncooked prawns and shrimp is presented in Figure 4. chilled 81 . The claws are usually bound together to prevent the animals fighting and causing damage to each other. they are washed. In comparison shrimps die quickly after capture.3 Processing and its Effects on the Microflora Crabs.1. Shrimp will then be glazed and frozen in a raw state. Scrupulous hygiene is important at this stage to prevent the cross-contamination of the shrimp tails. especially if they are stored in ice or iced water. Hence. Alcaligenes.. whether GHP are in place. Pseudomonas spp. As with finfish. by removing the head and thorax a source of enzymes is eliminated that will prevent the occurrence of black spots. chilled sea water. 4. The rate of spoilage will be a function of time and temperature. the rate of deterioration of the crustacean shellfish by these organisms is reduced. 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. from the time of capture and processing. These animals can survive out of water if the surrounding environment is kept cool and damp. or transported directly to processing plants. thus. Shrimp might be further processed with the shell on. When the shrimp arrive at the processing plant they are washed and then sorted into size.3. Micrococcus spp.1). gills and thorax.. there is evidence that the shelf life of tropical crustaceans is longer than that of temperate-water species stored correctly in ice . and will be dependent upon whether the crustaceans were harvested from cool or warmer waters. or are found in brackish water around the shores. As the muscles of crustacean shellfish decompose rapidly after death dead animals are rejected. Vibrio spp. bacterial contamination and spoilage are inevitable. graded according to size.
After cooking. Fig. and packed to meet the customer’s specification. 4. Flow diagram . 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. particularly the pathogens Bacillus cereus and Clostridium perfringens originating in the flour. spices and cooking batter. Good Manufacturing Practice (GMP). including GHP.frozen uncooked prawns 82 .FISH AND SEAFOOD frozen. should be followed to ensure that the product is not re-contaminated with microorganisms present in the processing environment or from the workers.1.
with the growing demand for ready-to-eat products from crab and lobster meat. However. wet and ‘soapy’ or ‘jammy’ to the touch. for crabs. If the tail drops. they include Sh. there is no associated spoilage problem. since crabs and lobsters are traditionally consumed immediately after cooking. or packaged and processed. Once removed and cleaned. The shells have a crisp.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. the meat is either repacked and sold in the shells. bright and dry appearance.4. slightly iodine smell. bacteria of public health significance. Heat is produced if the shrimps decompose in a confined space. 83 .3. there is a new hazard of contamination of the cooked meat by workers during its physical removal from the exoskeleton. with visible decomposition or green discoloration seen through the membrane exposed by the dropping tail. Processed crab and lobster meat can also be bought as packages of meat that have been subjected to a pasteurisation step. the joint between the tail and carapace is open and the limbs hang loosely. contaminating microflora may include enteric bacteria. The shell will be soft. spoiling prawns and shrimps have shells that are soft. In comparison. putrefaciens and Pseudomonas spp.4. the tightness of limbs will indicate whether they were killed by immersion in boiling water or. and have a sweetish. whilst boiled crustaceans have a pleasant slightly sweet smell. putrefaciens. 4.4 Spoilage The main genera of Gram-negative bacteria responsible for the spoilage of crustacean shellfish are the same as those that cause spoilage in finfish. dull and sticky.2 Cooked crabs and lobsters As indicated earlier. If boiled. and a strong ammoniacal odour is produced. immediately prior to boiling.2 Crabs and lobsters Crabs and lobsters should be alive when boiled or sold. viruses or parasites. 4.CRUSTACEAN SHELLFISH 4. With spoiled crabs and lobsters. there is no indication of life. 4. indicative of protein degradation and the growth of the principal spoilage bacterium Sh. or in hermatically sealed cans that have undergone an appropriate heat treatment. If hygienic procedures are not followed. An offensive odour is also produced.
1 Pathogens: Growth and Survival Salmonella spp. in 24. shrimp and shellfish (11). Although foodborne illness had not been associated with the product. monocytogenes in cooked ready-to-eat foods.5 4. The above results prompted the FDA to develop a compliance programme for the pathogen. cooked and smoked fish.5. Moreover. nearly 30 years elapsed before the potential hazard of L. in compliance with the Food and Drug Admistration’s (FDA) ‘zero tolerance policy’ for L. Aquatic birds and the hands of workers have also been identified as sources of the pathogen. However. animal manure. shrimps and cockles sold (17). and to increase testing of many other domestically produced seafoods under the General Pathogen Surveillance Programme (14. They are mesophilic microorganisms usually residing in the intestinal tracts of human beings and warm-blooded animals.2 Listeria spp. later surveys in 1990 recovered the pathogen from retail raw. and usually for both. 16). The presence of L. Results of a survey carried out in England and Wales between 1987 and 1989 demonstrated that L. One of the earliest accounts of the presence of Listeria monocytogenes in crustaceans was by Russian workers in 1959 (9). 7). fish fingers. An import alert was put in place in June 1987. 5. monocytogenes in contaminated crustacea was recognised in the United States of America (USA) when the pathogen was isolated from Mexican frozen cooked crabmeat (10).5% of frozen raw shrimps from different countries (13). which required automatic detention of all Mexican frozen crabmeat that tested positive for Listeria and Escherichia coli (12). even if GMP is followed. the first in a series of Class I recalls was issued in May 1987 to retrieve nearly 4 tonnes of tainted crabmeat (11). The pathogen was later identified in frozen raw shrimp (13) and lobster tails (14) . Following this. although efficient cooking will quickly eliminate it (6). a common pond fertiliser used in aquaculture. often contains Salmonella and other enteric pathogens (8) and has been demonstrated to contaminate crustaceans growing in these environments. 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. 6.cooked prior to consumption. monocytogenes in cooked ready-to-eat crustaceans is recognised as a potential risk to selected groups of individuals: the very young and 84 . Hence. in domestic/imported shrimps (15). Salmonella cannot be completely removed from raw shrimp during processing. The genus Salmonella contains a wide variety of ‘species’ pathogenic for man or animals. monocytogenes was absent from 40 samples of cooked prawns. as well as for Salmonella.FISH AND SEAFOOD 4. Further surveys identified Listeria spp. 4.5.
whilst boils and cuts are also common sources of the pathogen. 4. Crabs are covered in a chitin exoskeleton.5 Vibrio spp. 4. cholerae. which can become colonised by V. botulinum type E at 10 °C but not at 4 °C (20). is widely distributed in aquatic environments. although contamination by types A and B may occur from the environment. so is able to survive heat treatments that eliminate vegetative cells.70% of healthy individuals carry toxin-producing strains of Staph. especially in products that have been vacuum packaged. monocytogenes has been attributed to crustaceans. Researchers have demonstrated that inoculated shrimp supported toxin production by C. although more recently the non-O1 type has also been demonstrated to cause foodborne illness in crustaceans. However. 22) as well as from undercooked crustacea when harvested from contaminated waters (23. An epidemic that began in South America in 1991. 24. as part of the natural bacterial flora. Control should be achieved by promoting GHP and staff training programmes. To date. aureus on their hands. 19).CRUSTACEAN SHELLFISH old. cholerae non-O1. The pathogen has resistant spores. 4. cholerae is divided into two groups. aureus is not a member of the intrinsic flora associated with crustacean shellfish. the pathogen can grow at temperatures as low as 3 °C. which was eventually attributed to a ship discharging its bilge into the coastal waters of Peru. 85 . about 50 . pasteurisation and cooking. particularly crabmeat (18. combined with the degree of handling of these types of product. the causative agent of cholera.5. Vibrio cholerae.5. no incidence of foodborne illness caused by L. was associated with seafood consumption. e. 25). aureus because of the ubiquity of this organism in the processing environment.5. V. botulinum type E is commonly associated with fish and fishery products. V.g. cholerae O1 and V. Vibrio parahaemolyticus and Vibrio vulnificus are species that have been reported to be associated with raw crustaceans harvested from estuarine waters (21. GMP should be followed in the processing plant to ensure that the final product is held at 3 °C. The main source of contamination is the hands of processing personnel. pregnant women and the immuno-compromised.3 Staphylococcus aureus Staph. 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.4 Clostridium botulinum C. Crustaceans have been linked with food intoxications attributed to Staph. The former. Once the competing microflora has been removed by a heat treatment. therefore.
sobria are known to be fish pathogens and are commonly isolated from ponds used for aquaculture. South America. vulnificus is an invasive and lethal pathogen associated with wound infections. or freezing at a temperature of -23 °C for 7 days (30.5. Of these. a marine halophile. however. 4. Hepatitis A virus presents the most serious hazard. The organism is widespread in estuarine waters and is a common isolate from harbour water in warm climates. A. in a survey carried out in the United States between 1977 and 86 .8% of samples contained C. the potential hazard exists as C.5. 32). but has the potential to cause fatal foodborne illness (26). although A. parahaemolyticus.5. non-B Hepatitis viruses (31).8 Parasites The use of animal excreta in aquaculture has also been associated with foodborne parasitic infections of crustaceans. Caliciviruses. Japan.30 MPN/g) (27).g. V. 4. whilst 5.5. coli. Quantitative levels were below limits of detection in all cases (<0. 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. It is more commonly implicated in countries where crustacean shellfish are eaten raw. e. Norovirus. V.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. occurs in crustaceans from warm environments. to date. particularly trematodes (6). hydrophila and A. Astroviruses and non-A. been implicated in a case of foodborne infection associated with crustaceans.6 Campylobacter jejuni and Campylobacter coli C. Parasitic infections are controlled by either cooking infected material prior to consumption.9 Viruses Crustaceans grown in waters contaminated with sewage may become carriers of viruses such as Hepatitis A. 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. 31.7 Aeromonas hydrophila and Aeromonas sobria A. jejuni has not. However. they have not been confirmed to be causal agents of foodborne illness associated with crustaceans (28). 4. 4. However. jejuni was isolated from 36 (15%) of 240 samples of freshly hand-picked blue crab (Callinectes sapidus) meat. hydrophila is able to grow well at refrigeration temperatures (0 .
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. or chlorine concentrations similar to those found in drinking water. Selected parts from this regulation is presented below. Both Hepatitis A virus and Norovirus are stable pathogens and are relatively resistant to inactivation (34). The risk of such hazards is far higher with fin fish and molluscan shellfish.47% of outbreaks attributed to Hepatitis A virus were attributed to a foodborne origin (33). 4. They can survive low pH levels.6 Published Microbiological Criteria The global legislative requirement for controlling the safety (and quality) of crustacean shellfish is to apply a preventive.6.5. risk-based system based upon the seven principles of Hazard Analysis and Critical Control Point (HACCP) supported by pre-requisite or good practices programmes at all stages of the food chain. Norwalk viruses can also withstand heating to 60 °C. although it is one of the European Union (EU) legislated allergens. consistent with that in the stomach.10 Allergens Allergens have not been reported to be a significant hazard with crustacean shellfish. only 0.5.CRUSTACEAN SHELLFISH 1981. which results in a persistent and apparently permanent loss of short-term memory (36). A summary of microbiological criteria for crustacean shellfish is listed below. 4. 87 . Therefore any microbiological criteria need to be part of this HACCP-based system to verify that the system is under control.1 EU Legislation Microbiological standards for cooked crustaceans and shellfish. 38. 4. 39). The toxin causes amnesic shellfish poisoning (ASP) when consumed. No foodborne incidents have been attributed to this source to date. which could be transferred to the meat during unskilled removal from the shell (35). 4. from farm to fork (37. 2073/2005. Provisions laid down concerning the use of additives would also apply to these types of products. produced or imported into the European Union (EU). are included in the Commission Regulation (EC) No.
FISH AND SEAFOOD
126.96.36.199 Microbiological criteria applicable to the production of cooked crustaceans (Commission Regulation (EC) No 2073/2005 on microbiological criteria for foodstuffs) 188.8.131.52.1 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. 184.108.40.206.2 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. 220.127.116.11 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.
CURED. 93 . This can be called ‘kench’ curing and is mainly used for non-fatty white fish. rather than for preservation itself. Wet salting or brining. Simon Derrick The Grimsby Institute Humber Seafood Institute Europarc Grimsby DN37 9TZ United Kingdom 5. The processes take anywhere from a few hours to a few days or even months. dependent on the country or the type of product.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. The immersion in concentrated brines for long periods is usually used for fatty species and for longer-term preservation. their combinations and/or variations. and the extracted moisture is allowed to drain away. and marinating processes. fish is immersed for a few hours in a relatively low concentration brine. The fish is immersed in a brine solution in which different salt concentrations can be used according to the process. to assure a salt concentration in the fish flesh that is appropriate for flavouring purposes prior to the use of other preservation techniques. Fish can be salted in three different ways or their combination: Dry salting. It includes the salting. alternating fish layers with a layer of salt. and a preliminary operation to some smoking. drying. where the salt replaces moisture in the fish flesh. drying and smoking processes. split and opened out flat. Usually. Salting is both a method of preserving fish. Fish curing is practised worldwide. and is sometimes known as ‘pickling’. SMOKED AND DRIED FISH Dr. The fish is headed.5. both to preserve and to create a variety of new products. and salting is done by stacking the fish in layers. and before significant spoilage takes place.
Thus. without other preservation methods applied after boiling. It has found limited application with fish. the product rehydrates quickly but does not retain reabsorbed moisture on cooking. The principle of preservation of the first method is the inactivation of enzymes and killing the non-spore-forming organisms. thus reducing the water activity of the product. Included in this group are some lightly preserved products where no acid is added to the marinade. Vacuum drying. This is the common method of drying fish. There are some marinating processes where salt is used for the first cure and a firmer product is obtained. Fish in contact with refrigerated plates freeze. The fish is mixed with dry salt but in a watertight container. where an organic acid. Longer shelf life is achieved in the products when salt is added.FISH AND SEAFOOD Dry / wet salting. evaporated water is removed by vacuum pump. Food products can be dried in three different ways: Air or contact drying. The product is dried by contact with heated surfaces or by radiation. Smoking is an ancient process for preserving and flavouring food. such as ‘gravad’ fish. As the water is drawn out of the flesh by the salt a brine is formed. as concentrated brines are formed. are added to the marinade. Marinating or pickling. Drying is the preservation of fish by removing the moisture content of the fish flesh by evaporation. 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 . this was done naturally by exposure of the fish to the air. thus making the product unsuitable for microbial growth. Salt. or boiling it with dry salt and water. this ‘brine’ can be collected and placed again over the fish several times. usually vinegar. Salted-boiled fish are the products obtained on either boiling fish in a brine. Marinades are acidified brines. When the process is running in a closed system. Freeze drying. depending on the type of product. it can also be named ‘pickling’ or ‘blood pickling’. Salt-boiling. the ice sublimes and the vapour is removed from the fish by vacuum pumps. Usually. Traditionally. spices). prior to or during the boiling process. Heat is transferred to fish from heated air or a heated surface. is used as a preservative instead of salt. the preservative effect is due to the low pH used in the process. The products are usually consumed raw. and the air movement removes the vapour. as well as other flavourings (sugar. under normal pressure. and the fish can be immersed in such brines for long periods. It is not commonly used for fish products. thus delaying spoilage for a few days. After the freezedrying process. This is a mixture of a dry and a wet salting process.
the fish is subjected to heat for a sufficient period of time to coagulate protein throughout the fish. In this process. It can be injected into the product. 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. However. which impart the flavour of smoke to fish. 5. leaving only elevated levels of carbon monoxide. With the exception of some Asian countries. In this process. 95 . 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. sporeforming bacteria or osmophilic moulds. or added to the brine with other flavourings. Liquid smoke is a natural aqueous condensate of wood smoke that has been aged. Clear smoke. and the preservative effect is only slight. A dry or wet salting step is always used in the process before exposure to the wood or sawdust smoke. or liquid smoke. Also. Hot-smoking. The different spices used in the immersion brines are also a potential source of microorganisms. by exposure to the wood or sawdust smoke. such as injection or dipping in a solution of liquid smoke. The secondary objective is to impart sensory characteristics such as smoke colour and smoke flavour to the product. which irreversibly combine with the haemoglobin in the tissue to maintain the red colour.CURED. including pathogens. It can be smoked either traditionally. and filtered to remove tars and particulate matter. The process was designed so that the smoke is filtered to remove all smoke particles. by exposure to the wood or sawdust smoke. SMOKED AND DRIED FISH of smoke. or by means of smoke flavouring processes. the fish is subjected to heat for a period that does not coagulate protein. It is commonly used in place of traditional smoke. smoking is now used mainly to give the fish a characteristic flavour. or by means of smoke-flavouring processes with liquid smoke. Cold-smoking. Usually. After the salting step. The smoke itself may contain spores of moulds.2 Initial Microflora The initial microflora are those of the raw fish from which the products are made. the salt used in the processes may contain halophilic bacteria. Smoking can be either traditional. Usually. Smoked-flavoured fish. process temperatures are over 60 °C. the sugar used in some products may contain yeasts and moulds as well as spore-forming bacteria. temperatures below 30 °C are used.
The strength of the solution is described as degrees of saturation (brineometer degrees).3 Processing and its Effects on the Microflora Owing to the wide variety of products. When a 100º brine is used. only the most common processes can be described here.6 264. it is important to state the temperature. TABLE 5. Salt in the water phase links the two conditions of moisture and saltiness.80º brine (184.2 g of salt per litre of brine).2 105. or salt in the water phase. is employed.8 211. Thus.4 52. When working with salting.2 237. all using salting steps. Different brine strengths are required according to the curing processes and/or the type of product. At 18 °C a 100º brine is produced when the solution contains 26. it causes swelling because the fish absorbs water from the salt solution (1). Therefore. processes and producers’ private recipes. % 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.4% salt (Table 5. a 0º brine saturation means pure water. and salt content. For smoking salmon.0 158. the fish is usually salted with a 70 . about 50º (132.4 184.0 g of salt per litre of brine). This should be distinguished from the salt content.8 211. Salt in the water phase is the amount of salt compared with the total amount of water and salt in the fish. When a milder brine.FISH AND SEAFOOD 5.8 79.I). it is impossible to establish standardised processing stages in the curing of fish. which is the percentage of salt in the whole weight of fish that is water plus protein plus fat. smoking or marinating processes. The dissolution of salt in water is dependent on the temperature. A 100º brine solution means that the solution is saturated and no more salt can be dissolved.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) 26.0 . a 90º or above brine (237. sometimes the salt crystallises onto the surface of the fish. it is important to understand the concept of salt concentration. etc. Salt concentration means the percentage of salt in the water contained in the fish.6 132.6 g of salt or more per litre of brine) may cause salt burn in this way.
1 ppm and 10 ppm. It is common for the dried salted cod in the Mediterranean countries to be made using solar salts. lean white fish is used for this purpose. It takes about 3 weeks until proper salt penetration and moisture loss are achieved. Usually it is followed by other preservation techniques such as drying and/or smoking. However. 5.1. but the concentration of copper and iron must be kept under 0. This is because fatty fish turn rancid quickly. the fish is mixed with one part of salt per eight parts fish. Salting can be done in a number of different ways. The fish is then stacked in piles of about 2 metres and the water is allowed to drain away. For a light salting.1 Dry salting / Kench curing Usually. low in calcium and magnesium.35% and 0. and the salt has penetrated throughout the fish (3). and essentially free from iron and copper (3).CURED. SMOKED AND DRIED FISH 5.0. having a short section at the end of the tail. for which there is a high demand in Mediterranean countries. producing characteristic pink patches. are recommended to help make the product whiter. addition of levels of 0.3. mainly in tropical conditions. can be dried further to produce ‘klipfish’ or ‘bacalão’. and a ‘putty’ fish product is also obtained (1). ‘Salt burn’ might occur when the surface of the fish dries out too quickly as a result of the small grain size of the salt. The salt used should be of food-grade quality. are likely to cause hydroscopicity of the product and. 97 . This condition is known as ‘putty fish’ (4). ‘Green cured’ split cod with about 60% of the backbone removed.1 Salting Salting of fish for preservation purposes has been used since pre-history. caught in North Atlantic waters. which consequently inhibits the diffusion of the water from the bulk of the product to the surface. it tends to lead to a slight yellowing and softening of the product.3. however.15% calcium and magnesium. and such delays. the fish is restacked with fresh salt after about a week.15 . outgrowth of halophilic bacteria may occur. 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. On the other hand. Also. When pure salt is used. See Figure 5. if very pure sodium chloride is used for curing purposes. A ‘green cured’ product is obtained when the water content has been reduced to about two-thirds of the initial amount. when rock salt is used the ‘dun’ patches caused by moulds are prevalent (1). respectively. respectively (1). and salt can speed this oxidation process. can lead to the decomposition of the centre of thick muscles before it reaches the level of salt concentration sufficient to prevent spoilage. when the air humidity is high. and for a heavy salting a ratio of 1:3 salt:fish is used (1). The most common fish used in the northern hemisphere is cod (Gadus morhua). The calcium and magnesium impurities. which include calcium and magnesium.1.
eviscerated and/or split). Only small fish can be salted whole. such as 98 .FISH AND SEAFOOD Dry salting can also be used for small species without the need for opening the fish flat. For large fish it is advisable to split it before salting. The greatest barriers to salt penetration are the skin and scales. Processing of dried salted cod 5. so that the risk of spoilage is high.2 Wet salting or brining As referred to above. Fig.5.90 are rarely achieved. although toxin production would be unlikely (4).3. Food-poisoning organisms. Where fish are salted in their whole form. the process is mainly for flavouring purposes prior to the use of other preservation processes. it appears that water activities much less than 0. when a light brine is used. and the uptake of salt is dependent on the type of fish (fatty or lean) as well as its presentation (whole.1. such as Staphylococcus aureus could survive and grow at such water activities in the product.1.
41 3.90 (1). sardines.84 1.56 6.5 3.5 3. limiting the amount of rancidity that can develop.45 3.78 2.23 2.23 3. For example. the fish enzymes play an important role by maturing or ripening the product.17 4.44 2.46 4.76 99 .88 5. Here.70 6. and a red burgundy colour will develop in the product.64 4.11 9.78 6.5 4.33 5.04 5. after being drained and pressed at 465 kg/m2 for 15 hours.11 4. but as it is placed in containers. herrings and other fatty fish can be used.96 1.19 3. 5.41 6. but mackerel.99 2.17 5.73 1. Fresh sardines. SMOKED AND DRIED FISH smoking.3 Dry / wet salting In this process.76 4. Histamine formation is of importance when using scombroid and some clupeidean species.06 4.47 6.09 8.95 3. This is why it is the preferred method for fatty fish. they can be stored at 28 °C. the fish is dry-salted.30 2. There are several variations of this process.66 5.3 days.61 3.1.98 6.03 4.98 5.CURED.86 2.80 5.23 8. the product obtained is called gaspé-cured (5). a saturated brine is used.3.0 3. the brine is then replaced over the fish.85 3. with a shelf life of about 4 weeks (water activity (aw) about 0.5 10.98 6.76 4. thus. For preservation purposes.0 2. can be cured in a saturated brine solution for 6 days and. with pressure being applied to the top of the container to help the brine to drain away. the water that comes out of the fish forms a brine in which the fish stays immersed for 2 .61 3.74 9.26 1.60 2.09 8.10 1. Anchovies are the main fish used for these purposes.03 2.65 2.44 2.25 5. The brining step for smoking purposes also helps the product to develop a desirable glossy appearance. Histamineforming bacteria do not generally grow at water activities below 0.00 3.11 3.51 5. See Table 5.66 5.70 3.26 7. The main objective of the brining method of salting fish is to limit air penetration during processing.26 4.22 6.91 2. TABLE 5.26 3.76 4.41 4.23 2.49 4.54 5.76 4. The fish are kept immersed in their own brine near saturation point for several days.05 7.II.76 4.35 4.II Salt in the water phase (after Ian Doré (5)) Moisture (%) 40 45 50 55 60 65 70 75 80 85 90 Percentage salt in the flesh (by weight) 2.78) (1).26 2.51 3.25 5.85 3. the liquid that comes out of the fish forms a brine. if split fish is dry-salted. It is then taken out and dried in the sun or in kilns until about 35% moisture in the product is obtained. and can be collected and replaced again over the fish.41 5.16 7.0 4. headed and gutted.
then packed in barrels with a 70º brine. flavouring (sugar. However.3. Once cooled. In a second step. and 6% salt. Herring can also be cured for several days in an 80 to 90º brine with 2. a Peruvian product. The fish can be soaked in a high concentration of vinegar (about 4 .3 Marinating / pickling Fish can be brined in solutions based on organic acid instead of salt. After this. underlines the need for careful control (4). The cooked portions are poured into bowls containing gelatin dissolved in water (10% solution) and sometimes a small amount of vinegar. Another variation uses sour cream instead of the second brine step. the occasional implication of pindang in cases of foodborne illness. giving a cooked appearance to the product. until the flesh is soft. only the nonspore-forming organisms are eliminated in the process and. They will have a salt content >6% (w/w) in the water phase. As referred to above. placed in a marinade of lime juice. outgrowth of bacterial spores after processing may occur (4). and even death. or can be boiled by being placed in a basket. and packed (3). Ceviche. Another salt-boiled Indonesian product.3. Traditional jellied eels are a product where pieces of fish are cooked in boiling water. The first process can give a longer shelf life. Afterwards. because of the salt content of the product. usually flavoured with onions and hot chillies. which is dipped into a boiling brine for 10 .5% 120grain distilled vinegar. salt. The Indonesian ‘pindang’ is made by boiling the fish in a saturated salt solution and it is sold as is. The product is usually consumed raw with the high strength acid solution softening the bones enough for them to be eaten. usually requiring refrigeration below 10 °C for longer-term storage. they are packed for consumption within 2 weeks at chill temperature (6).2 Salt-boiling Fish can be boiled after being dry-salted in a watertight container and heated for at least 1 hour. has a shelf life of several years.30 minutes.2% acetic acid and 2 . 5. There is epidemiological evidence that these products have been the cause of some 100 .2 months. drying and salt-boiling steps until a hard translucent block is obtained (4). they can be cut and cured for 3 days in a solution with 3% white distilled vinegar. is made of different varieties of fish. up to 1 . The lime juice plays the role of the vinegar. and lasts for 3 . Usually. even if the product is hermetically sealed. or a pH <5 and they are stabilised for a limited period. whereas the latter method preserves the fish for only a few days.4% of salt) for distribution and sale (1). Other preservation techniques can be used for extending the shelf life of these products. herring or other fatty species are used.FISH AND SEAFOOD 5. A much firmer product is obtained when the fish is salted prior to acidification. ‘ikan kaju’. and spices) is added to the brine.6 days without refrigeration (1). All these products are ‘semi-preserves’. but it has further smoking. it can be repacked in milder brine (1 .7%) for as long as 3 weeks. containing 1 kg of salt for every 25 kg of fish.
the fish is dried naturally in a relatively cold environment. split and dried gadoid species) are traditionally made in cold climates. then mild-cured with special seasonings and packed in barrels with 80 º brine (5). Dried octopus is also a very common dish in the interior of the country.There are several products named marinades (matjes. and the stacks are placed in the refrigerator. and a fine product is obtained. gutted. Matjes are made of fat young herring. but sometimes it is dried without prior salting. mixed with salt. after frying.) where no acid is added to the process. Other products. the temperature and the amount of pressure applied (5). The most common method used for fish products is the air-drying process. Usually. Fresh dill is placed between the layers of fish.5. 101 . At pH levels below 4. It takes about 2 days to marinate completely. South America and the Caribbean. In the first case. air flows naturally. 5. bacterial toxins and parasites (7). a very fine delicacy for Oriental food lovers. are generally air-dried at sea. by using either solar heaters or mechanical dryers.3. which are placed in a mixture of salt. Provided the product is kept in dry and fairly cool conditions. sugar and spices. depending on the thickness. They are not cooked prior to consumption and they have a limited shelf life even at refrigeration temperatures. are likely to survive in these mild-processed products. Stockfish products have a golden brown colour and are very hard. it being preferable to use frozen salmon or to freeze the product at the end of the process to avoid parasite survival. such as squid. Some bacteria and enzymes are still active in the presence of acid and salt. It is important to assure the quality of the raw material. are sundried and. while in the latter case there is a provision for fans or other means of mechanically moving the air (1). marinated products will keep for several months at a temperature of 4 °C. the fish is dried after salting or brining. Dried shrimps are widely used in Asia. Stockfish (dressed. sea cucumbers and seaweeds. still made without prior salting on a large scale. gravad fish. and the flesh will break down. etc. are commonly dried in Asian countries. They are an important dried product. Stockfish is extensively used in Portugal. In some coastal areas of Portugal.CURED. under a light pressure. SMOKED AND DRIED FISH foodborne diseases related to the presence of biotoxins. 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 >5.0) (7). Gravad fish is a Scandinavian product made of salmon fillets. In Scandinavian countries. 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). it will last for many years (5). used as an appetiser.4 Air drying This is probably the oldest method of fish preservation. jellyfish. some fish species such as moray eel and varieties of dog fish. Foodborne pathogens. Shark pectoral fins. such as Listeria monocytogenes and Clostridium botulinum. Italy and Spain (5).
After dry salting. the fish must be dried slowly enough to avoid case-hardening. nor subsequent cooking operations will affect the presence of toxins (e. Whilst the production of dried fish products is tightly controlled in industrialised countries. the ‘green cured’ cod is rinsed or washed with water and then hung to dry either by the sun and breeze or. some of which are listed in Table 5.FISH AND SEAFOOD When the fish is sun-dried in tropical conditions. However. Italy and South America. skin up. the quality is more difficult to control. 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. 102 . spoilage and possibly C. and the water must be changed several times before cooking. C. making a water barrier and consequently. 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. by warm air circulating in a drying chamber. To prevent this. Thus. that may have contaminated the product during the drying process. Spain. neither it. tarpaulin. toxic secondary metabolites of the moulds Aspergillus flavus and Aspergillus parasiticus are not produced at aw <0. dried salted cod (bacalão) is now mainly produced in Norway. botulinum growth might occur in the bulk of the product. Portuguese and Spanish fisheries cannot satisfy their own countries’ demand for cod. botulinum). They must be soaked in water. the product can be held for months at room temperature. aureus) or spores (e. there is an increasing demand from people that have emigrated to industrialised nations for traditional products. Owing to restriction of fishing areas for Spanish and Portuguese vessels since the 1970s. France. The fish must be stored in reasonably dry conditions to avoid moisture absorption. the better the moisture transfer. Staph. and increasing amounts of such products are making their way into the market place. Although controls are in place to ensure that products processed in such relatively uncontrolled conditions do not enter the supply chain.83 (9)). The amount of moisture transfer from the fish to the air is dependent on the relative humidity .the lower the humidity. the protein must be set or denatured with low-temperature drying before applying higher temperatures. In conclusion. but fast enough to avoid spoilage caused by bacterial and enzymic activities (1). Dried salted products must have a water activity below 0. if the rate of water removal is too high in comparison with the diffusion rate. The product is dried to a final water content of about 25 to 38% (3).III. or wooden drying rack to dry in the sun. more commonly nowadays. which leads to mould growth (aflatoxins.g. Traditional processing conditions provide ample opportunity for product contamination from a wide range of hazards and sources. Iceland and Canada. typically in stews and soups. the skin will form a hard case (case-hardening). It is important to balance the drying rates and quality/safety issues. A highly demanded dried fish product in Mediterranean countries is dried salted cod.75 (1).g. In many such artisanal fisheries the drying is simply conducted by spreading the fish on the beach. and it is exported to Portugal. When no more water can be removed from the fish. and has the primary objective of creating a product with specific taste and textural characteristics.
Proper cleaning and preparation prior to smoking and washing of the fresh or frozen fish by water spray or a continuous 103 .salmon. botulinum (spores) Pesticides / Chemicals Physical Glass / sand etc. 5. The smoking procedures vary widely. It has been reported that the smoke from mahogany. is mainly a process for flavouring fish rather than a preservation technique. cod roes. Aroma and flavour are a blend of smoke components (3). making full description impossible here. Staph. tuna. Preventing the occurrence of carcinogens during smoking has been an issue for the past several years. Maintaining the temperatures of pyrolysis between 425 and 200 °C. softwood colours the product quickly. mackerel. Blends of various woods can be formulated to produce sawdust with specific flavours. It can be used for cold or hot smoking. such as Escherichia coli. eel. haddock. and the use of smoke generated by superheated steam or liquid smoke. aureus and Saccharomyces cerevisiae (10). cod. white mangrove and abura has an inhibitory effect on some microorganisms. The quality of the smoke is related to the type of wood used. temperature and humidity can be automatically controlled and the process is faster because the directed air movement draws the smoke through the fish.3. Top-quality fish must always be used. However. The production of smoke by wood or sawdust burning can be done in a mechanical kiln or in traditional chimney kilns. There are many fish species that are used for smoking purposes . sprats. Hardwoods are considered better because of the milder flavour they impart to the food. herring. as referred to above. trout.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. mussels and many others from tropical countries.CURED. using electrostatic filtration of the smoke. aureus (toxin) C. oysters. SMOKED AND DRIED FISH TABLE 5. are some of the ways of reducing the polycyclic aromatic hydrocarbon (PAH) compounds (10). In the mechanical kiln.5 Smoking Smoking today. clams. swordfish.
If wet salting is chosen. skin off or on. they can be vacuum packaged. skin on or off). spices. filleted. The fish can be dry-salted using a weight of salt 5 . 104 .10% of the weight of the fish. Brining at 10 15 °C is recommended (10). Sugar. whether cold. All the smoked products.5. smoked cod fillets and finnan haddock are usually cooked before consumption. or packed in retortable foil pouches. all cold-smoked products should be cooked before consumption. When frozen/thawed fish is used.1 Cold-smoking The major difference between cold. it will reduce the salty flavour. If frozen fish is used. wooden boxes or master cartons (10). after being smoked need to be chilled immediately to 4 °C before being packed. as well as many other products prepared in the same way. are boiled before smoking (12). such as smoked salmon-trout. tuna and swordfish. but a ‘safety’ (i. and the cold smoke is denser. shrink packaged. In cold-smoking. as it retains more moisture. but this concentration will again depend on the type of product and the length of the brining stage. pasteurisation) temperature is not achieved in any stage of the process.or hot-smoked. or quick-frozen and stored at -18 °C (2). Some herring products. However. it will double the drying time in the kiln to get the same total weight loss as for dry-salted fillets (11). canned or bottled. an attractive glossy pellicle on the fish fillet surface is achieved after draining. The length of the smoking process for such products is much greater than for the hotsmoked fish. the following step is salting.FISH AND SEAFOOD water flow system with chlorinated water (25 . it will take up salt twice as fast as fresh fish (10). Sugar can be added together with the salt. the highly desirable cold-smoked salmon. temperatures must be kept under 30 °C throughout the process. Wet salting or brining is preferable to dry-salting when smoking fillets because. Cod roes. thus allowing the deposit of smoke on the fish surface. This difference is crucial for safety issues because the flesh is not cooked. giving the product the desired appearance and flavour (4). The salting period is dependent on the size of the fish and its presentation (whole. Also. 70 – 80 º brine is commonly used. thawing must be done slowly for 12 . low temperatures will slow the uptake of salt.e.50 ppm) should always be done just before process initiation (3). which is important for the appearance of these products (4). but it will increase towards the end of the process (10). Whether the product is prepared headed. 5.and hot-smoking is the temperature of the process. temperatures and times used in processing coldsmoked fish are very favourable for the proliferation of food-spoilage and foodpoisoning microorganisms. usually dry-salted and cold-smoked. Thus. gutted or filleted. owing to the elution of soluble proteins. Drying should be minimal at the start. however. covering the fish surface completely (10). The maintenance of temperatures below 30 °C avoids protein coagulation. permitted colourings or smoke flavours can also be added to the product in the brining step. Depending on the type of product. Thus. the excess salt is removed with water.18 hours at 4 °C. are not cooked prior to consumption.3.
further cooled to 3 °C within 12 hours and then packed. Length of the smoking process at each stage will depend on the species and the type of product. the products are cooled. botulinum type E the product must have at least 3% salt in the water phase and be stored below 5 °C. Afterwards. as. there is no point in the process that can fully assure the absence of L. A typical temperature set is as follows: firstly. 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. These products should be vacuum packed at room temperature. botulinum type E present an even longer time than before to grow and produce toxin (14). Vacuum packing extends the shelf life of the lightly smoked products by suppressing the growth of other bacteria and moulds. are of concern when smoking fish from tropical waters (13). Hot-smoked products require a longer cooling stage.2. they are more sensitive to sodium chloride and unlikely to grow in smoked products (13).3. can be isolated from seafood. such as Yersinia enterocolitica. but they should be cooled to a temperature of 10 °C within 3 hours. a partial cooking period at 50 °C. Smoked products can be also packed under controlled-atmosphere packaging. in order to produce toxin. thus allowing any psychrotrophic C. if they are taken from a chill room at 3 °C and packed. Neither the smoking temperature nor the salt content is enough to kill the organism. To avoid growth of non-proteolytic C. which underlines the danger of storage at abuse temperatures of >10 °C (13).CURED. Pathogenic vibrios.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. but the proteolytic types A and B. can grow with salt levels of 10% in the water phase. Vacuum packing is recommended only for coldsmoked salmon. gravad salmon (also called gravad lax or gravlax). 5. refrigerated storage and vacuum packaging still allow its growth (12). If the refrigeration chain is maintained. an initial humidity of 70% seems to be preferable because it is a good compromise between maximum smoke adsorption and minimum casehardening (1). if present. a drying period at 30 °C. SMOKED AND DRIED FISH During cold-smoking. monocytogenes. On the other hand.5. Smoked salmon fillets are usually vacuum packed whole or after slicing. condensation will occur inside the pack. if a 105 . and hotsmoked salmon (kippered) (3). secondly. typically mesophilic. and their sensitivity to salt (13). The protein must be set or denaturated with lower temperatures before the application of higher temperatures to avoid case-hardening. The exception is vacuum packed fish. During smoking. a final cooking period at 80 °C (12). stored at 4 °C and packed. See Figure 5. Although other human pathogens that can grow at refrigeration temperature. and the recommended gas mixtures are 60% carbon dioxide and 40% nitrogen (15). thirdly. Furthermore. This temperature should be maintained during storage and distribution (3).
it will have a reduced shelf life. and moulds will grow on the product (12). 5.FISH AND SEAFOOD product is packed too warm without being cooled to room temperature. Fig. Process of cold-smoked sliced salmon 106 .2.
Hot-smoked products made from white fish will usually keep better than those from fatty fish. a very expensive and fashionable smoked product in the Netherlands. Natural smoke extracts. Boiled or steamed clams. The classical liquid smoke is 107 . Vacuum packaging retards the onset of rancidity when fatty fish is used. Eels. such as yeast derivatives. apart from the improved appearance.3. In terms of safety. A type of dried hot smoked tuna called masmeen or masmin is produced in the Lakshadweep islands of the Indian Ocean. 22. The composition of commercial liquid smoke can vary widely depending on the method of manufacture. Growth of these moulds on the tuna results in the extraction of moisture. They are generally not cooked prior to consumption. The product is removed from the drying cycle. 21. then hot smoking and finally sun drying it. resulting in a characteristic flavour (20. are usually hot-smoked after being immersed in 80º brine or dry-salted (6). including ‘umami’ (16. 5. Hon kare bushi is a form of Katsuobushi from which shavings are cut to make aromatic stocks for use in Japanese dishes (24). Katsuobushi. rewetted and remoulded to give the desired shape and consistency. are usually 80º brined for 5 . cooking in brine. and many closely related Aspergilli (Aspergillus ochraceus.CURED.3 Liquid smoke Smoke-flavoured fish represents a small percentage of the total smoked fish industry output.3½ hours using the typical temperature set for hot-smoking referred to above (12). chilled or freeze-thawed. SMOKED AND DRIED FISH There are several species that are usually hot-smoked and the process can vary markedly. 17. they can be stored for 6 months at -30 °C and for even longer when frozen in vacuum packs (12).6 hours and smoked for 3 . are used. 18). Aspergillus ostianus. and substances unconnected with smoke but with a smoky flavour or smell. (Penicillum cyclopium and Penicillium puterellii) (19). 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. but seems. The production method is similar to masmeen. including Eurotium repens and Eurotium rubrum. Aspergillus oryzae. hot-smoked fish can be classified as a ‘heat-treated or pasteurised’ product (7). synthetic smoke flavours. Mackerel. white fish about 8 days. Aspergillus tamarii) and several Penicillium spp. moulding it into long thick sections.5. break down of fats and the moderation of smoke flavour. 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). When refrigerated at 3 °C. Masmeen is usually prepared from bonito tuna by eviscerating and filleting the fish. 23). but mechanical drying is used instead of sun drying. fatty fish will remain in good condition for 6 days. 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. They will keep well frozen. The product is often allowed to go mouldy by incubating it in a chamber containing a natural fungal flora.
06 μg/kg of benzoanthracene in the final foodstuff (28). both of which are considered potential carcinogens.FISH AND SEAFOOD smoke condensate that is dissolved in water or oil. so as to impart an unnatural red colouration to the flesh. 5.4 Spoilage Brining. established in its guidelines that smoke flavourings added to the food product must assure a level lower than 0. or applying it as an atomised spray within a modern automatic smoke kiln. the salt level could effectively be reduced from 4. Liquid smoke was considered to provide advantages of safety.5. In experiments comparing traditionally smoked fish with liquid smoke.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. There are two groups of chemicals of concern in smoke: polycyclic aromatic hydrocarbons (PAHs) and nitrosamines. It has been shown that liquid smoke can reduce the formation of PAHs (3). The European Scientific Commitee for Food. Smoke-flavoured fish processed using liquid smoke can be prepared by including liquid smoke in the brine. spraying is preferable because it is easiest to control flavour and acceptable with this method (3). applying it as a dip after brining.8% and still provide protection for 7 days (25). injecting it into the fish flesh. 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. 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. consistency of flavour and colour.6 to 2. and less than 0. 5. 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. or smoke extracts in organic solvents. drying. it has been reported that the liquid smoke-treated product has less pronounced aroma and flavour (25). Fresh fish spoilage may be measured by 108 . smoking. According to some recent research. pickling and marinating fish were originally used to lengthen the short shelf life of fresh fish. Whilst neither the CO or the carboxymyoglobin complex are harmful in themselves.03 μg/kg of benzopyrene. 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. 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). nutritional neutrality and antioxidant activity (27).3.
otherwise.. for some. The use of drying and/or salt to lower the water activity. Even saturated brines allow some halophilic organisms to grow. for others. the consequent reduction in the pH will help to inhibit spoilage organisms. but. marination or drying). Once placed into brines. Some brines are re-used over a period of days and a load of 10. pickling and dry-salting all introduce bacteria to the fish. Pickling and/or the anchovy process uses solid salt extraction of fish fluids to form a brine solution (cure). the pickle and dry-salting preservation processes require further processing for significant shelf life increases (e. When heavy brining or salting is important for product shelf life. The microflora attached to the fish is relatively unaffected by light brines used before smoking. the shelf life may be estimated and investigation made into the causes of the loss of sensory quality. Brining. contact with the bacteria 109 . and high concentrations are usually required to inhibit their growth fully. which may also be produced to describe the quality attributes and their loss over time for processed fish. Most marinated and smoked fish products are first brined or salt-pickled. the fish flora are transferred to the brine mix. solar salts investigated carried the highest load and included halophilic organisms as a result of their production method (open-air evaporation of sea water). as the fish are added. and coryneform bacteria. Micrococcus spp.000 cfu/ml of brine may build up. and of smoke to add chemical preservatives. Once these changes are defined. chemical oxidation. Within the limits of safety. a process similar in microbiological effect to that of saturated brining. Stronger and longer brining has an effect like that of pickling. the bulk of the microflora comprises Bacillus spp. mainly comprising corynebacteria and Gram-negative bacteria from the fish and water (29). therefore.g. In salts. the production process will modify the microbial flora present on the product.. When acid marinating or smoking. these halophilic organisms may grow and cause spoilage. or their products require refrigerated storage. For some products. rock and manufactured salts did not usually contain halophilic bacteria (29). Upon salt packing.3%).CURED. the fish are first cleaned (known as ‘rousing’) by a salt rub to remove any outer slime. a product may become undesirable for consumption as a result of changes in its sensory characteristics. Usually. or extended refrigerated storage. Salts used for brining have been found to carry a microbial load of between 10 and 1000 organisms per gram of wet salt. both of these will affect the spoilage patterns. however. This is not usually the case for smoked fish as the residual salt concentration is not high enough to inhibit all the non-halophiles. SMOKED AND DRIED FISH grading schemes or quality indices. Endogenous marine fish bacteria prefer to grow in the presence of salt (marine concentration approximately 2. contaminant bacteria from other materials used to produce the product. spoilage is due to endogenous microorganisms and enzymes of the fish. also. limits the types of microorganism capable of growing. which are found in the salt. Often. a very dry product is required to produce 'shelf-stability'. and. Spoilage needs to be carefully defined for each product. the fish are cut and splayed to allow greater surface contact with the salt. these bacteria tend to continue to dominate the flora in the liquid.
34) have shown that P. gaspé). flavus. Several Penicillium spp. niger. as the water activity of saturated brine solutions is low enough to inhibit most bacteria. although it may occur in other salted fish.g. Eurotium amstelodami. and Halobacterium spp. powdery growth. or after drying if the fish is stored in excessive humidity. Sporendonema sebi)). rubrum are also common isolates from dried fish. The bacteria form coloured colonies on the surface of salted fish prior to drying. most notably Penicillium citrinum and Penicillium thomii. halophilica was isolated 110 . lactic acid bacteria (LAB) and moulds. including hydrogen sulphide (26). rancidity due to lipolysis. The microbiology of heavily salted products is dominated by Gram-positive. together with a range of Aspergilli. The bacteria are osmosensitive and will lyse in water. yeasts and fungi. yeasts. if not too contaminated. protein breakdown. A. as little if any growth will occur below 5 °C on salted fish (29). ‘dun’ is not characterised by the production of offensive odours. Only halophilic microbes may grow. including A. spore-forming bacteria. though only the latter two species were considered likely to have grown on the fish (32). sometimes covering the whole of the fish with a whitish. Aspergillus wentii. Bacteria from these two genera also grow sufficiently in brine and even salt to cause pink discolouration. (syn. A number of spoilage organisms can grow and produce conditions such as ‘sliming’. sebi is considered to be the main spoilage agent of dried fish produced in temperate regions (30). Basipetospora halophila. At the pre-drying stage. like the ‘pink’ organisms. or fat breakdown) caused by enzymes rather than by microbial growth. 32.FISH AND SEAFOOD in the salt is made and the flora will develop accordingly in the blood pickle. 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). E. 33). it was found in about 50% of samples examined. ‘Pink’ describes a condition associated with surface growth of the pigmented halophilic bacteria Halococcus spp.. B. the best way of controlling the organisms is to refrigerate the fish. and still produce a reasonable dried product. spoilage of these pickled and dried fish in refrigerated storage tends to be due to chemical changes (textural changes due to proteolysis. ‘Sliming’ or ‘putty’ fish is a condition that occurs in poorly or insufficiently salted cod (e. repens and E. and Aspergillus penicilloides. pisce was the main fungus isolated. it is possible to wash off the infected parts. The metabolism of the organisms will produce off-odours. Insufficient salt penetration into the flesh leaves areas where Gram-negative rod-shaped bacteria and micrococci grow and cause a putrefactive spoilage. may also be found in the salt used in production. Aspergillus sydowii. which. This condition is often associated with case-hardening. were also isolated. The condition describes the growth of the halophilic moulds (Wallemia sebi. W. Therefore. (syn Oospora halophila) and Polypaecilum pisce are two white halophilic moulds that are associated with spoilage of salted fish (31. halotolerant or halophilic micrococci. Studies conducted on dried and salted fish from Indonesia (31. Their growth produces noticeable spoilage of the fish appearance but is also controlled by refrigeration. Unlike ‘pink’. ‘pink’ and ‘dun’.
so the bacteria spoiling these products will be psychrotrophic or psychrophilic. The heat during hot-smoking will kill vegetative microorganisms but not all spores. but often colour changes due to chemical oxidation of the smoked product will limit the shelf life. which does not create the salty conditions required for spoilage by those halophilic microorganisms. and microbial spoilage is more likely. Aspergillus clavatus and Aspergillus niger.IV..CURED. The brining to 3 . but. In Vietnam. as already stated. Vacuum packing limits chemical oxidation. Fish processed by cold-smoking. or any smoking process where temperatures in the interior of the fish do not reach >50 °C for a significant period of time. 111 . it was. however. so the most probable spoilage organisms will be spore-formers or post-smoking contaminants. but this will not prevent the growth of spoilage bacteria. Smoked fish products tend to have a light brine treatment. The salt in the water phase must be high enough to inhibit the growth of C. LAB.747. able to grow on dried salted fish. not the growth of spoilage organisms. Marine vibrios were the dominant flora in smoked salmon that was drysalted. SMOKED AND DRIED FISH infrequently at low levels. flavus. 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. 33. where a high internal temperature is reached. Reports for vacuum packed cold-smoked salmon have identified more than one organism associated with spoilage. Cold-smoked fish will contain microbial flora representative of that of the raw fish. The preservative effect of the smoke will inhibit some spoilage microorganisms. will have a different residual post-smoking flora from 'hot-smoked' fish. Technical studies demonstrate the ability of many of the xerophiles above to grow at typical water activities seen in dried salted fish (32. Refrigeration of hot-smoked fish will greatly reduce the number of organisms able to grow and will lengthen the shelf life. whereas a mixture of Enterobacteriaceae and LAB were prevalent in brineinjected salmon (37). Pseudomonadaceae. 36). Vibrionaceae and Enterobacteriaceae may all play some part in shelf life limitation.5% sodium chloride in the water phase will inhibit some bacteria.V gives details of pH ranges and aw minima for the growth of some important spoilage microorganisms and pathogens (38). including A. It has been reported that the type of salting (dry. Aspergillus spp. with a lower aw limit for growth of 0. For more information on spoilage see Table 5. Refrigeration is necessary. has been isolated from dried fish (35). around the water activity of saturated sodium chloride. botulinum type E. Table 5.
W. marinated and smoked fish Spoilage controlled by pH Heat Ant1 Ref2 Process Product Aw Probable spoilage organisms None Fresh fish spoilage organisms Halophilic spore formers and PPC4 Halophilic spore formers and PPC Halophilic organisms: Pink-Halococcus spp. Matjes v3 + ++ v - +++ ++ ++ - v +/- ++ ++ ++/++/- Pindang Bacalao + +++ Gaspé cure Anchovie ++ Anchovies ++ - - - ++ ++ Saturated brined fish Salted then boiled Salted. Histamine producers Halophilic spoilage organisms Spore formers and PPC None Acid-tolerant spoilage organisms Xerophilic moulds Fresh fish spoilage organisms. Enterobacteria and PPC Spore formers and PPC Variable Salt marinades Salted then boiled Brined then boiled Salted then dried Canned Fish Gravad. halophila Halophilic spoilage organisms Halophilic spoilage organisms. 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.not used/unchanged. sebi and B. LAB. . + to +++ strength of preservative effect 4 Post-process contaminants 112 .variable. boiled.IV Spoilage of salted. and Halobacterium spp. Dun.FISH AND SEAFOOD TABLE 5.
8 1. popular demand for healthy foods has led to a change in the tastes preferred by the public and. Before. aureus Yeasts Moulds Lactic acid bacteria pH 5. although the combination of the two factors.86 0. botulinum type E and production of toxin. 3% sodium chloride in the water phase is enough to inhibit growth of type E for at least 30 days (39). A number of surveys have shown that fish from temperate waters is mainly contaminated with the psychrotrophic C. Sodium nitrite in conjunction with sodium chloride is known for its ability to inhibit C. SMOKED AND DRIED FISH TABLE 5. respectively. with this change. botulinum. but if the fish products are stored at temperatures <10 °C. types A and B are not affected by 3 . The presence of salt in cured products has a great effect on the growth of the bacteria. In the United States. under optimal conditions.75 0.5 0-11 3.V pH ranges and aw minima for growth of microorganisms (38) Organisms Most Pseudomonadaceae C. the non-proteolytic type E strains can grow in up to 5% sodium chloride. However.5-8. pressure on the traditional methods of curing. 9 and 8 of the outbreaks.5 5. botulinum type E Staph. Examination of 165 outbreaks of botulism caused by fish products showed that smoked.5% of salt in the water phase and great care must be taken to avoid any temperatures >10 °C (13). sodium 113 .97 0.5% to 5%.5.1 Pathogens: Growth and Survival Clostridium botulinum In the last few decades.8 0. botulinum Staph. aureus Most spoilage bacteria Most spoilage moulds Halophilic bacteria Xerophilic moulds Aw 0. the importance of cold-chain integrity must be emphasised. salted and pickled products accounted for 10. could not grow and produce toxin. It has been shown that. the fermented products being the most dangerous group (7).61 Organisms Most Pseudomonadaceae C.9 0.97 0. and as botulism is a serious hazard in foods. botulinum type E.0-9.5 4.5-10. The concentration required to prevent growth at room temperature can vary from as low as 3. but the presence of the mesophilic types A and B is of concern in fish from warmer waters (13). smoked and dried.5-8. the products were heavily salted.5-10 4.CURED. temperature ≤5 °C and salt ≥3% in the water phase is sufficient to prevent any growth of type E. when present in the raw material. Since neither the smoking nor the drying parts of this curing process are particularly severe.5 5. although approximately 10% salt in the water phase is necessary to fully inhibit the mesophilic types A and B strains (14). and C. but the concentration of salt in smoked salmon or trout (usually varying from 1 4% salt) is not high enough to prevent growth.
Usually.2 Staphylococcus aureus This is a ubiquitous organism but its main reservoir and habitat is animals and. the organism can survive most of the light curing processes.0% salt in the water phase (30). As with other organisms. if recontamination occurs and storage conditions are favourable.94 (7). monocytogenes is a ubiquitous organism that has been detected in fresh fish skin. Shigella and E. if the person handling the product is a carrier of Staph. but.4. The human carrier rate can be as high as 60% of healthy individuals with an average of 25 .3 Enterobacteriaceae The pathogens Salmonella. gills and guts as well as in several lightly preserved fish 114 . if storage temperatures are over 10 °C. when the bacterial load of the product is lower.5. An outbreak of Shigella in an Italian cruise ship. they do not grow well in the raw material. In foodstuffs.0 . its occurrence in a final product is due to contamination during processing by an infected asymptomatic carrier with poor personal hygiene. 5. However.4 Listeria monocytogenes The psychrotrophic L. Sliced smoked fish products. the human nose. more importantly. aureus growth. they can survive lightly cured processes when the sodium chloride content is less than 5% and water activities are higher than 0. has been reported (42). it should contain not less than 3. The FDA established that hot. 5. The degree of concern in coldsmoked salmon is low unless temperature abuse >10 °C occurs (13). throat and skin. Infection by Shigella sonnei has increased in the UK. are particularly susceptible to contamination and. but. Thus.5. related to the consumption of a raw smoked fish meal. as they require more handling. It is a halotolerant bacterium able to grow at water activities as low as 0.86 and 10% sodium chloride.or cold-smoked fish should have at least 3. if it contains sodium nitrite ≥100 ppm. Once the product is contaminated. but requires higher temperatures for toxin production (>10 °C). coli all occur on fish products as a result of contamination from animal/human reservoirs. they will grow rapidly (7). The organism is mesophilic with a minimum growth temperature of 7 °C.30% of the population positive for enterotoxin-producing strains (41). As they are poor competitors. growth and toxin formation may occur.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. 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. and the minimum pH for growth is 4. the risk of infection can be eliminated by proper cooking.5. 5. after processing. aureus.5. the organism may contaminate the product.5% salt in the water phase.
Until 1997. monocytogenes pathogenic for only specific segments at risk.the round worm Anisakis spp. are considered unsafe. or nematodes. 5. Marine LAB is part of the natural flora of chill-stored vacuum packed salmon and cod. monocytogenes in levels at or above 102 per gram (46). Pseudoterranova dicipiens. which considered L. monocytogenes’ in 25 g of the finished product. This includes matjes herring. both being detected in cold-smoked salmon.5. salt levels of 8 .92 . in the EU Fisheries R&D-FLAIR program. which. a round worm. pH 4. Its limiting conditions for growth are 0 . They have complicated life cycles. Unlike some European countries. The use of LAB in fish preservation is a topic that has been studied in projects (UP. Some LAB. the CT 3162-FAIR program.2.514. such as ‘lightly preserved’ fish products. SMOKED AND DRIED FISH products such as cold-smoked fish. gravad fish. the only concern to public health is when eating raw or uncooked fish products. which include intermediate hosts. a short period of freezing either of the raw material or of 115 . a number of fish products. have been shown to have an anti-listerial activity in many food products. lightly salted caviar. and in CT95-1207FAIR). and other local traditional products (7).9. the USA established ‘zero tolerance’ for the organism. Thus. As they are heat-sensitive.0.95 (45).5 Parasites Despite the presence of parasites in fish being very common. Of microbiological concern in the northern hemisphere are two parasites . 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.6 and water activities below 0. when an outbreak of nine cases of listeriosis was suspected to have been caused by gravad or cold-smoked trout (47). The round worms.45 °C. has also been detected in cod (7). As freezing to -20 °C and maintaining this temperature for at least 24 h will kill all the nematodes. which means that the process must conform to a level of ‘no detectable L. and the tape worm Diphyllobothrium spp. cold-smoked fish. widely used as starter cultures in food processes. which means that cooking a fillet 3 cm thick for 10 min at 60 °C will kill any worms present (48). researchers are looking for other ways of preventing the pathogen’s occurrence or growth. favours growth of psychrotrophic LAB. from seawater fish. most of them are of little concern to public health.8 . there were no reported outbreaks related to cured fish. Vacuum packaging (VP) of sliced smoked fish creates a microaerophilic atmosphere.CURED. from freshwater fish. marinades (ceviche) (43) and gravad fish (44). in combination with curing agents and refrigerated storage. As its presence has been reported in lightly preserved fish products whose processing chain cannot fully assure the absence of the organism. do not survive 1 min at 60 °C. Two outbreaks in the United States have been reported to be linked to salmon (10).12%.
<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. taramasalata and cooked shellfish sampled at the point of sale (adapted from (11)) Criterion Satisfactory Borderline limit of acceptability (cfu/g unless stated) Unsatisfactory Unacceptable/ potential hazard* (cfu/g unless stated) (cfu/g unless stated) Aerobic plate count† (30 °C.100 >100 N/A Pathogens Salmonella spp. Acceptability is based on appearance. † Guidelines for aerobic colony counts may not apply to certain fermented foods.<104 20 . coli (total) Listeria spp.<107 ≥107 N/A <100 <20 100 . (not L.VI Guidelines for the microbiological quality of ready-to-eat smoked fish.<100 200 . E. salami. cholerae Vibrio parahaemolyticus L. and the levels or absence of indicator organisms or pathogens.<100 20 . texture.<100 10 . not detected in 25 g Campylobacter spp. monocytogenes Staph. for example. the organism should be identified # N/A denotes not applicable 116 .FISH AND SEAFOOD the final product should therefore be included in the processing as a means of controlling parasites (7). cereus and Bacillus subtilis# <20 <20** <20 <10 20 . monocytogenes) (cfu/g unless stated) <106 106 . soft cheese. 48h ± 2h) Indicator organisms‡ Enterobacteriaceae E.<104 104 .<103 N/A 100 . perfringens B.<100 >104 >100 N/A N/A <20 20 . These foods fall into category 5. 5. and unpasteurised yoghurt.<104 100 .6 Published Microbiological Criteria TABLE 5. If the Bacillus counts exceed 104/g. ‡ On occasions some strains may be pathogenic.<100 20 . aureus C.<104 present in 25 g ≥103 >100 ≥104 ≥104 <103 103 . smell.
Catching. 117 . 2. 82 (revised). 1993.. London. 1984.. Torry Advisory Note No. FAO Fisheries Technical Paper. 7.W. Wareing P. New York. 120-38. Van Nostrand Reinhold. in Micro-Facts: The Working Companion for Food Microbiologists. Martin R. Chapman & Hall..M. Regenstein C. 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. cured and dried fish. 9. in Fish Processing Technology. Torry Advisory Note No. Leatherhead. where 30 ppm is permitted Paralytic shellfish poisoning* Amnesic shellfish poisoning * *These criteria are applied to all fish 5. Eds. 1997.. References Regenstein J. 3273. monocytogenes Salmonella spp. Leatherhead Publishing. 4.. 56. USA.CURED. New York.H. Food spoilage fungi.F. 1991. 381-406. parahaemolyticus Vibrio vulnificus C. or presence of toxin 0. Horne J. McLay R. 1979. The Smoked and Cured Seafood Guide.8 ppm (saxitoxin equivalent) 20 ppm domoic acid. Doré I. Handling and Processing Eels. Assurance of Seafood Quality. except in the viscera of dungeness crab. 1994. 6. salting and smoking). 37(revised). Fernandes R. coli (ETEC) L. Hall G. Ed. Moody M. Wareing P. in The Seafood Industry. Torry Advisory Note No. 3. Smoked. Marinades. 1990. SMOKED AND DRIED FISH TABLE 5. Van Nostrand Reinhold. Horner W. * Staph. Bannerman A.A. Birnie K. Flick G. Huss H.VII US Food and Drug Administration (FDA) guidelines related to ready-to-eat fishery products (45) Product Ready-to-eat fishery products (minimal cooking by consumer) Organisms and/or toxins Enterotoxigenic E. 334. Hot Smoking of Fish.7 1.F. Fernandes R.E. Preservation of fish by curing (drying. Introduction to Fish Technology. Eds.M. Flick G. 1982. 254-82 5. botulinum * Limit of tolerance 1x 103 ETEC/g. Urner Barry Publications Inc. aureus * Vibrio cholerae V.. 8. 2007.
Yajima I.B. 843... and Biochemistry.. Lancaster. 16. Bioscience. Patel T. Bannerman A. Halpern B. Lindemann B. Bioscience.K. 1980. 1984.... Food Control. Processing Cod Roes. Packing Fish in a Modified Atmosphere.. 1994. 53 (4). Technomic Publishers. 1386-7. Agricultural and Biological Chemistry. Biotechnological Applications. 24.C. Ninomiya M.. Perusing Asian soup options. 2324-5.. 23. Food Product Design. Bioscience.. Flavour constituents of dried bonito (Katsuobushi). 17. 1989. The discovery of umami. Doi M.. 6 (4). Paranjpye R. Shiemori Y.. Matsui M. Teeny F. Cann D. 2008. Kanayama T. Ed. Bannerman A.. 27 (9).. 5 (revised). London. Matsui M. Ninomiya Y.A. Effect of dried bonito (Katauobushi) and some of its components on GABA-A receptors. 19. Chapman & Hall. Peterson M. Torry Advisory Note No. Inhibition of Clostridium botulinum types A and E toxin production by liquid smoke and NaCl in hot-process smoke-flavoured fish. Jeppesen V. Doi M. Journal of Food Protection. 1977. Control of biological hazards in cold smoked salmon production. Torry Advisory Note No. 18. Horne J. Fish Drying and Smoking: Production and Quality. 22 (revised)..C.. Druschitz A.FISH AND SEAFOOD 10..M. Food Chemistry.. 958-60. Doe P. 6-dimethoxy-4-methylphenol by Aspergillus glaucus and their possible contribution to Katsuobushi flavour. 11. Asymmetric reduction of acetophenone by Aspergillus species and their possible contribution to Katsuobushi flavor..M. 2002. 59 (12). Ogiwara Y. 21. Degradation and omethylation of phenols among volatile flavour components of dried bonito (Katsuobushi) by Aspergillus species. Shuto Y. 15. Botulism and Fishery Products. 1992. 1997.F.. 25. Hosokawa M. Pelroy G. Torry Advisory Note No. in Fisheries Processing.R. Dillon R. Eklund M. 17 (9). Aoshima H.. 1051-5. Doi M. 840-6. 108 (3). Torry Advisory Note No.. Yokayama S. Martin A. Microbiological control for fish smoking operations. Kinoshita Y.M. Martin A. Hayashi K. Recommendations for the Preparation of Smoked Salmon. 1990. 508.H. 2007. 22. Shuto Y. Oxidation of acetophenone by Aspergillus species and their possible contribution to Katsuobushi flavour. Huss H. 335-40.W. 6 (6). 13. 88. 1998. O-methylation of 2. 935-41. 1981.. 56 (6). 51-81. 118 . 12.. 20.E. 1995.. 18. 1995. Yamauchi H. Sakakibara H.. 553-72. 14. Kinoshita Y. Cann D. Biotechnology and Biochemistry. Shuto Y.P. Mitou M. Doi M. Biotechnology.E.. Food Reviews International. What’s in a name? Are MSG and umami the same? Chemical Senses.. Embarek P. 1982. 61 (8). 45 (10). Biotechnology and Biochemistry.
Ed. Pitt J.I. Borgstrøm G.S. Journal of Applied Bacteriology. Food Technology.E. 1986. Taylor L. 107-11. Sprouse R. 34. 1985.O. Hocking A.. Hocking A.1988. Adebajo L. 74... 1996.B.. 40. Anggawati A. Fuchs R. ceviche. 119 . Incidence of Listeria monocytogenes in an acidified product. Hocking A. Importance of autolysis and microbiological activity on quality of cold-smoked salmon. 1941. 32. Hocking A. Lucas F. 123-9.D. Golden D. 70-2. 1961. Wheeler K. in Fish as Food.. 36. Fungi and Food Spoilage. Hansen L. Nahrung. 27. Studies on salt fish. 14.Y.I.M..R. 49 (1). New York. Oxford.. 29 (2)... Cox J. 164-9.. in Fish and Fishery Products Hazards and Controls Guide. Hess E.. 218-25.J. Fungal flora of South Vietnamese fish and rice. The microbiology of sea-water fish. Pszczola D. Smoked and smoke-flavoured fishery products. Sirvas S. Journal of Food Technology. 33. Blackie. Interactions among xerophilic fungi associated with dried salted fish. 1991.C. Fungi associated with Indonesian dried fish. 31. Frank M. Letters in Applied Microbiology. 12. Loessner M. 1995. Luxembourg.T. Journal of the Fisheries Research Board of Canada. Modern Food Microbiology. 1994. CRC. 88-90. 5. Jay J. Diyaolu S.D. V. Committee on Evaluation of the Safety of Fishery Products. Huss H. Academic Press. Studies on Sporendonema epizoum from ‘dun’ salt fish. 39.A. National Academic Press. 311-7. 1994..K.A.V. Effects of sodium chloride and relative humidity on growth and sporulation of moulds isolated from cured fish. 1995. 42. Maga J. in Smoke in Food Processing.. Institute of Medicine. 348.F. 74 (2).. 742. 43. 1997. Boca Raton. 3 (4). Scientific Committee for Food.I. Pitt J. CEC.D. London. Seafood Safety. Springer Science. Food Research International. 1996. in Reports of the Scientific Committee for Food (34th Series). Tour highlights production and uses of smoke-based flavours. Ed. 38. Gill T. Cann D. 37.. 28. 1993. SMOKED AND DRIED FISH 26. Lancet.F. Journal of Tropical Medicine and Hygiene. Food and Drug Administration. The control of botulism in hot-smoked trout and mackerel. Røntved S. Food and Drug Administration. 41.A. 351-7. 98100. 30. Wheeler K.. 197-208. Press Inc. 1971. An outbreak of Shigella. 1991.. Pitt J. 35.M. Mycotaxon.E. Ed. Smoke flavourings... 1979. 27686. New species of fungi from Indonesian dried fish.M.H. 181-8. Ed.A. 74. Simini B. Commission of the European Communities. Washington DC. 22. Rockville. Ahmed F.D. US Food and Drug Administration. Antimicrobial properties of smoke. Townsend J. 38 (3).CURED. 2005. 29. Shewan J. Food Microbiology.
1992.. Journal of Clinical Microbiology. Round Worms in Fish. The effect of NaCl on the upper temperature limit for growth and enterotoxin synthesis by Staphylococcus aureus. Danielsson-Tham M-L. 1991. 58 (8).L. 1994. Hurst A.M. Amsterdam.8 Further Reading Gibson D. 135-85. Roberts D. Growth and production of toxin of Clostridium botulinum type E in rainbow trout under various storage conditions. 13-8. 1980. 2000. 26 (4).C. and smoked fish establishments good manufacturing practices. Food and Drug Administration. Davies A.html Gilbert R. Acta Veterinaria Scandinavia.V. Hughes A.. Hobbs G.A.A. Donovan T. Incidence and growth of psychrotrophic Clostridium botulinum in food. Pathogenic microorganisms of importance in seafood..W..M. 188 (3). Liston J. 45.A.cfsan.. 120 . 47. 55 (4). Journal of Food Protection. Persson I. 1989.S.. Eklow A... Advances in Food Research.. Torry Advisory Note No. Fletcher R. 863-6.FISH AND SEAFOOD 44. 1996. Gibbs P. 1995. Nye K.. Peterson M.gov/~comm/haccp4x5. 1980. 3.. Susuki E.. US FDA Centre for Food Safety & Applied Nutrition. 232-6.W. Harrison M. 49-59. Prevalence of Listeria monocytogenes and other Listeria spp.. Appendix 5: FDA and EPA Guidance Levels... 46. 197-209. de Louvois J.N. Inhibition of Clostridium botulinum types A and E toxin formation by sodium nitrite and sodium chloride in hot-process (smoked) salmon.J. Elsevier Science Publishers B. 1997.. An outbreak of listeriosis suspected to have been caused by rainbow trout. Tham W. in smoked and "gravad" fish. Bolton F. Garren D. in Quality Assurance in the Fish Industry.C..R.... Cann D. Eds. Microbiological guidelines for some ready-to-eat foods sampled at the point of sale. Communicable Disease and Public Health. Loncarevic S.. in Fish and Fishery Products Hazards and Controls Guide.. Paranjpye R. Unnerstad H. Journal of the Association of Food and Drug Officials. Mentzing L-O. Association of Food and Drug Officials.D. http://www. salted. 5 (1)...fda. Pelroy G.. Huang Y. Huss H. Tham W. Ribeiro C. Little C.. 35 (11) 2904-7. 2001. 48. 5-7. 45 (9).J. Richards J. Canadian Journal of Microbiology. 833-41. Food Control. US FDA Centre for Food Safety & Applied Nutrition. 1982. Ericsson H.H. Beltran A. Ed. 163-7.. Clostridium botulinum and its importance in fishery products. Zeitschrift für Lebensmittel-Untersuchung und –Forschung.. 22. 5. Wooten R. Danielsson-Tham M. Moral A. 1976. Pelaez C. Cured.. Eklund M. Loncarevic S. 80.. Keeping quality of vacuum packed smoked sardine fillets: microbiological aspects. 37 (1). Jacobsen M. 507-10.E. Washington D. Journal of Food Protection.
Roberts T. Letters in Applied Microbiology.CURED. Slade A. Applied Microbiology... solute. temperature. Applied Microbiology.178-80. detection and growth of Listeria monocytogenes in seafoods: a review. 19. Poindexter C. 1992. 1994.. Histamine production by psychrophilic flora. Eds. 1987.. Hitchins A. Ben Embarek P.P. Journal of Food Protection. Contamination pattern of Listeria monocytogenes and other Listeria spp. SMOKED AND DRIED FISH Sutherland J. Ratnam S. Elsevier. Lashbrook L. Jemmi T. Davies A.J. Eklund M. Interacting effects of pH.. International Journal of Food Microbiology.A. Nicolaides L. 844-6. Taylor S. 21 (3). Combined effects of water activity.. Fate of Aeromonas and Yersinia on modified-atmosphere packaged (MAP) cod and trout. 293301. Liston J. 4389-95. 94-8. in Seafood Quality Determination. Fuchs R. Peterson M.R. Incidence of Listeria in hot and cold-smoked fish.. Tran T. Marine toxins of microbial origin..R. Pelroy G. Occurrence of Listeria monocytogenes in freshwater farms and fishsmoking plants. Dillon R. 1995.S. 55 (11).. Letters in Applied Microbiology.R. 42. Prevalence of Listeria in smoked fish..T. Kramer D. Dallas H.C. Yndestad M. 1991. pH and sodium chloride. Journal of Food Protection. 1995. Food Microbiology. Beuchat L. Predictive modelling of growth of Staphylococcus aureus: the effects of temperature. and temperature on the growth of Vibrio parahaemolyticus. 1995... 121 ... Competition of food bacteria with Listeria monocytogenes during enrichment culture. 502-8. Rørvik L.W. 25. Beuchat L. 11..E. Hudson J. 61 (12).. Keusch A. 1973.K. 27 (6). Bayliss A. 1988. Paranjpye R. 1974. 1734. 866-70.A. 23 (1). 14.. 1992. 309-16. 309-18. Efficacy of high sodium chloride concentrations for the destruction of Listeria monocytogenes. New York. 394-6.A. and salt concentration on growth and survival of Vibrio parahaemolyticus. Caugant D. Presence. 19-27. Patel T.. Poysky F.N. van Spreckens K. 25 (5). International Journal of Food Microbiology. Journal of Food Safety. Incidence and sources of Listeria monocytogenes in cold-smoked fishery products and processing plants. 58 (5). 217-36. 1994. Letters in Applied Microbiology. 1075-80. in a salmon slaughterhouse and smoked salmon processing plant. 11. Food Technology. 1994. 1994. International Journal of Food Microbiology.
FERMENTED FISH Prof. The fish sauces are produced on the largest scale.I.25% and are used mainly as a condiment. whereas in those where a carbohydrate source such as rice or sugar is also added.6.1 Definitions As applied to fish products. the term ‘fermented’ describes a spectrum of processes that ranges from the largely autolytic degradation of fish protein. the most important of all fermented fish products. North American and other markets and are. in economic terms. Essentially. 123 . Martin Adams Faculty of Health and Medical Sciences University of Surrey Guildford Surrey GU2 7XH United Kingdom 6.1 Fish/salt products such as the fish pastes and sauces tend to contain relatively high levels of salt. the best-known of the fish sauces (gyoshoyu) of Japan is shottsuru. Carbohydrates are added to some fish sauces during their preparation. Sugar can also be added to the Indonesian fish sauce bakasang and the Thai product nam-budu. typically in the range 15 . there are numerous variants of some common themes and a host of local names used to describe them. Fermented fish products are largely confined to east and south-east Asia. to those processes in which the activity of lactic acid bacteria (LAB) plays an important part. producing the fish sauces and pastes. 6. they are exported to European. The relative importance of these two activities depends on the formulation of the product. they can be divided into two categories: fish/salt products and fish/salt/carbohydrate products. The principal fish sauces and pastes of south-east Asia and their local names are listed in Table 6. For example. added during preparation. koji.1. It is produced from the sandfish Arctoscopus japonicus and has the rice-based enzyme preparation. lactic fermentation can play a significant role. though some are produced elsewhere. As with most traditional products that are still produced principally on a cottage industry or domestic scale. Those to which only salt is added tend to be mainly autolytic.
124 . who had adopted them from the Greeks. fish sauces known as liquamen or garum were ubiquitous condiments for the Romans.1. may be direct descendants.III. 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. a variety of products can be produced within a single country. 6. In Europe.2 Fish/salt/carbohydrate products range from those that resemble the fish sauces and pastes in the sense that extensive autolysis has occurred. salty taste. Fermented fish products are of very minor importance outside Asia. made from fish livers in Greece. to products analogous to other lactic fermented foods such as salami. A selection of Asian products is presented in Table 6. These have long since fallen from use. made from whole fish in France. and the more widely known Worcestershire sauce a very distant relative. where the bacterial production of lactic acid is a major feature. This is well exemplified by some of the fish/salt products of Thailand described in Table 6.I Fish sauces and pastes of south-east Asia Country Sauce Amber/brown liquid. and pissala. 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.II.FISH AND SEAFOOD TABLE 6. although garos.
som-fak/ som-fug 125 Malaysia Philippines Thailand . mam-pa-kor.g. shrimps. 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. pla-chao. e. pla-som. crustaceans Variety of fish Variety of brackish or marine fish 1:4 3 . pa-chao. the mixture is then boiled and bottled - Condiment Nam-khoei Shrimps.12 months 10% raw sugar added. som-pa-keng pekasam. burong-ayungi. tai-suchi phaak.III Fish/salt/carbohydrate products of south-east and east Asia Country Japan Kampuchea Korea Laos Products I-sushi. burong-bangus pla-ra. funa-suchi.4 months Condiment and main dish Whole 1:3 18 months Condiment Whole 1:3 After 3 . crustaceans Mackerel eviscerated fish Variety of fish Whole 1:4 Condiment Pla-thu-khem Whole 1:3 2 .g. pa-khem.II Fermented fish/salt products common in Thailand Name Type of fish Form Approx. som-pa.8 months Main dish TABLE 6. mam-chao. ayu-sushi.3 months Main dish Tai-pla Bowels 1:3 6 . burong-dalag. cencalok burong-isda. mam-seeing sikhae som-kay-pa-eun. e.FERMENTED FISH TABLE 6.
and old descriptions of their production often refer to the addition of whey.FISH AND SEAFOOD In Northern Europe. and a large local freshwater reservoir (2). and their microflora tends to reflect their local environment more than that of marine species. it was found that several different sources were used for the fish . Here. states that sugar is often included with the salt. 6. malt or flour (1). and the initial microflora is likely to be considerably larger in these cases. paddy ponds beside rice fields used for collecting fish when the field has dried up. in a survey of fermented fish products in north-east Thailand. rakefisk or surfisk. and counts ranging from below 104 /g to in excess of 107 have been reported. The diversity of fish used in fermented products (see. A modern account of their production. made from herring. 126 . especially in fish/salt products. relatively mild-tasting product. however. TABLE 6. the most popular varieties of which are rakørret. This could lead to quite marked variations since the same fish can be obtained from a variety of sources. freshwater fish are the usual raw material. There are also more heavily fermented products. in Norway and surstrømming. 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. is a traditional. Penaeus indicua Many of the fish used in fermentation are uneviscerated. for example. some fermented fish products remain popular. in Sweden. For example. which employ lower levels of salt than Asian products. fermented trout.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.the flooded rice fields. These are often described as fish/salt products. or buried salmon.IV Fish species used in lactic fermented fish products of the Philippines Product names Burong ayungin Burong bangus Burong dala Burong gurami Burong hito Burong kanduli Burong tilapya Balao balao Burong hipon Local Ayungin Bangus Dalag Gurami Hito Kanduli Tilapya Tagunton Suwahe English Silver perch Milkfish Mudfish Goramy Catfish Sea catfish Tilapia Shrimp Shrimp Scientific name Therapon plumbeus Chanos chanos Ophicephalus striatus Osphronemus goramy Clarias batrachus Arias manillensis Tilapia nilotica Macrobrachium spp. Norwegian gravlaks. Table 6.IV) and in the way in which they are handled prior to processing means that the initial microbial levels are far from uniform. which has been increasing in popularity over the last 50 years.
however. 6. the product contained about 500 cfu/g comprising mainly Micrococcus and Bacillus spp.3. were found to be the predominant isolates. where the total counts dropped from above 107 cfu/g to below 103 cfu/g after 14 days and below 102 after 40 days. angkak (Philippines and Thailand) and look-pang (Thailand) are used. These preparations contain a mixed microflora of moulds and yeasts. Studies have shown that the number of bacteria declines rapidly during the production of the Thai fish sauce nam pla and that. Hansenula. (3. It also demonstrated their significant proteolytic activity during the first month of fermentation. and studies 127 . which included nam-pla and patis. 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. probably reflecting their ability as spore-formers to survive rather than any capacity to multiply under the prevailing conditions (5).3 6. however. 4). In another microbiological study of four fish sauces. they can contribute to the initial microflora.4. principally species of the genera Rhizopus. More recently. It has also been shown that a perfectly acceptable patis could be produced from fish that had been sterilised with ionising radiation (6). In some cases. work with nam-pla in Thailand has isolated Halobacterium and Halococcus spp. mean that microorganisms have been generally held to play little or no role in the process that follows. Similar results were seen with patis. its contribution is likely to be insignificant compared with that of the fish microflora in terms of both spoilage potential and public health concerns. 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. either directly or as the partially saccharified rice they produce. Endomycopsis and Saccharomyces . in many cases. and the anaerobic conditions that prevail in the bulk of the material. Similarly microbial inactivation by irradiation has not been found to adversely affect the properties of Korean fermented squid (7). suggesting that halophilic bacteria do play an important role in the production of fish sauce (8). Other halophilic species have been identified from fish/salt products.1 – 6.74 or below. When this is a simple carbohydrate source such as sugar. traditional starter culture preparations such as koji (Japan). sufficient to saturate the water phase present and produce an aw of 0.FERMENTED FISH Where other ingredients are used. or boiled or roasted rice.1 Processing and its Effects on the Microflora Fish sauces and pastes Detailed flow charts for several fish/salt fermented products are presented as Figures 6. after one month. Bacillus spp. Generally. in numbers reaching 108 /ml after 3 weeks of fermentation but declining thereafter. Mucor. the Philippine fish sauce.organisms that play an important part in the production process but can also contribute to the ultimate spoilage of the product. This.
FISH AND SEAFOOD have demonstrated the production of halophilic proteinases from fish sauce isolates (9 . Fig.1. Patis (Condiment) 128 .2. 6. Belacan (Condiment) Fig. 6.12).
and muscle cathepsins have been shown to be inhibited at relatively low salt concentrations (13.4. To produce semi-solid fish pastes as opposed to fish sauces. Myulchijeot (Staple food) The extensive proteolysis and liquefaction that occur during production are.3. In fish sauce production. however. eviscerated fish are sometimes used to slow the autolytic process. 6. thought to be largely the result of autolytic breakdown of the fish tissues. 14).6. This is more rapid when whole fish are used since the head and viscera contain higher concentrations of proteolytic enzymes than other tissues.FERMENTED FISH Fig. Ika-Shiokara (side dish) Fig. the 129 .
8. filtered and bottled. for example.6 – 6.V and Figures 6. 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. particularly glutamate. Hoi-Malangpu-Dong (side dish. Some examples of products and their formulation are presented in Table 6. Their main feature is that the provision of a source of carbohydrate allows a lactic acid bacterial fermentation to occur. snack) 130 .FISH AND SEAFOOD high salt level used also assists in the osmotic extraction of a supernatant rich in amino acids.5.5). 6. 6. Fig. In contrast to free-swimming fish.2 Fish/salt/carbohydrate products of Asia This category encompasses an even greater diversity of products than the fish sauces and pastes.3. and the low pH produced contributes a second protective barrier or hurdle against the growth/survival of the normal fish-spoilage organisms and pathogens. which is eventually separated from the residual fish tissues. Figure 6. volatile fatty acids and nucleotides.
25-1 Fish:salt:roasted rice: garlic 10:1:3:1 Fish (minced):salt:rice: garlic 10:0. NE Formulation Fish (whole or pieces): salt: roasted rice.12 months Shelf life Use 1 .7 days 5 . the prefix 'pla' is replaced by 'kung'. C NE. south of Thailand. NE 10 . Kung-Chom (staple food. N.V Fish/salt carbohydrate products of Thailand Name 1 Pla-ra Region C. N.5:2-3:1 Fish:salt:boiled rice: ang-kak 3:1:3:0. north.5-1. central.3 years Condiment and main dish 2-3 months 3 weeks 2 weeks 2 weeks 6 .FERMENTED FISH TABLE 6.12 months Main dish 2 Pla-jao C.03 Preparation time 6 . Ang-kak: red mould rice obtained by fermenting rice with Monascus purpureus. N S 5 .2-4 Fish (pieces): salt:khaomark (fermented rice).6. C. Fig. 3:1:1-3 Fish:salt:boiled rice: garlic 10:2:1:0. 3:1:0. side dish) 131 .10 days 5 days Main dish Main dish Main dish Condiment and main dish Note: Products 1 . north-east. NE.6.12 days 3 .20 days 3 Pla-som 4 Pla-jom 5 Som-fak 6 Pla-paengdaeng NE. C NE. N. S.4 are also produced using shrimps. In these cases.
It also indicates that.10%. while they are able to grow at high salt levels.salt and lactic acid. 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. Phillipines Journal of Food Science and Technology. the salt is of far greater importance in determining the product’s long-term stability. Generally. these products are salted at lower levels than the fish sauces and pastes. This is readily explained by the susceptibility of LAB to elevated salt concentrations.3. then this will take longer to develop than if lower salt levels are used. their growth and metabolism are nonetheless slowed. A survey of fish/salt/carbohydrate products in Thailand (15) noted that there were two empirical rules governing formulation:6. typically in the range 2 .7.FISH AND SEAFOOD The extent to which lactic fermentation occurs depends largely on the salt content. If acidity is a desired sensory property of the product. The LAB isolated from these products are those with more marked salt tolerance but.6. (from Arroyo et al. Red burong isda 132 .2.VI). giving a reduced rate of acid generation. 106) Fig.1 The use of higher salt levels results in a longer production phase but a better keeping quality product. but there is considerable variation in this.. 2. of the two factors contributing to shelf life .
6.17.2 4.0 .3 years 1 .2 years 2 weeks 2 weeks Source: A Concise Handbook of Indigenous Fermented Foods in the ASCA Countries (16).9 .1 .6 3.4.2 4.5 ..8 .5.6.VI Shelf lives reported for fish salt carbohydrate products Product Pla-som Pla-paeng-daeng Pla-ra Pla-chao Pla-chom Som-fak Salt 18.104.22.168 pH 4.3 . 106) Fig.9 22.214.171.124 .FERMENTED FISH (from Arroyo et al. White balao-balao TABLE 6.5 3.8 2.0 Shelf life 3 weeks 6 .12 months 1 .3 5.0 .5 .5.9 4. 133 .5.0 .9.5.4 .9. 2. Philippines Journal of Food Science and Technology.1 4.2 7.8 .
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. 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). alternatively. The effect of this will be to decrease the product’s final pH and increase the amount of total acidity. they increase the amount of soluble sugars produced and thus the range of LAB that can grow. such as salt content. Rice is the most common carbohydrate source 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. TABLE 6. When added as boiled or roasted rice alone. although the final population of LAB may be similar.5) despite other factors. of course.1 .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. Addition of carbohydrate will. being similar (18.9) compared with the white product (pH 4. a saccharifying agent such as koji (Japan) or ang-kak (the Philippines) may be added separately (Table 6. the rice may be added in a partially saccharified form such as kao-mark (Thailand). This results in a more active lactic fermentation and a lower pH.VII Fish/salt/carbohydate products in which saccharified rice or a saccharifying starter culture is used Product red burong-isda i-sushi.3. also serves as a valuable source of the more readily fermented carbohydrate.2. also lower the unit cost of the product and this is probably a significant factor in many cases. a common ingredient in these products. In several products. inulin. 19).4. 21). Although the ability to ferment starch is not widespread in the LAB.3.VII). 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 . Garlic.FISH AND SEAFOOD 6.0 . Where they are used. efficient lactic acid production will depend upon the presence of amylolytic LAB or saccharification by other microorganisms or enzymes that may be present. For example.
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). One study found that any effective measure to exclude air from a fermenting fish/carbohydrate mix prevented yeast growth (23). Traditional packing techniques used with lactic fermented fish products. the level of salt employed is on the high side for fish/salt/carbohydrate products.FERMENTED FISH Sikhae. Any loss of acceptability must arise from chemical or physical changes occurring over prolonged storage.17.9%. Some authors have claimed that the function of the roasted rice added during pla-ra production is to improve texture and slow the fermentation rather than to serve as a substrate for lactic fermentation (22). In a successful fermentation. Cleaned pieces of eviscerated fish flesh are salted with 6% salt overnight before mixing with cooked millet. presumably correlating with the salt content.5 in the first 2 . red pepper powder and garlic in the ratio fish:millet:pepper:garlic of 80:7. thereafter declining to about 4. The mixture is allowed to ferment for 2 . such as wrapping in banana leaves. Clearly. and there is a marked regional variation in the preferred type. The growth of yeasts and moulds could. lead to an increase in pH.3 days. It seems that a lactic fermentation takes place in many types of pla-ra but is not significant in others.7 up to 6. but they are likely to behave similarly to lactic fermented products produced from other commodities. 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.2. which ranged from 7.5:9:3.2 at the end of fermentation. reporting the product’s pH to range from 4.3 weeks at around 20 °C.5. 135 .8 . is something of an exception since millet is used along with garlic as the carbohydrate source. which is sufficient to prevent lactic fermentation by all but the most halotolerant LAB.7 to below 4. the Korean fermented fish product. Excessive growth and acid production by the LAB could render products too sour for acceptability. will not reliably exclude air. but in most cases it is probable that yeast and mould growth would be an important cause of spoilage. Results of a product survey were consistent with this. around 12%. allowing other spoilage organisms to grow. the salt levels in the latter are sufficient to rule out microbiological spoilage in all but the most exceptional of cases. This suggests that use of oxygen-impermeable wrapping films could extend the shelf life of such products. 6. and surface mould growth has been shown to be a cause of rejection in the Thai product som-fak (24). Generally. There is little published information on the factors that limit the shelf life of the low-salt products. The Thai product pla-ra exemplifies these basic rules as well as the difficulties associated with attempts to classify such products. in turn.4 Spoilage The ASCA handbook on fermented foods in the region ascribes shelf lives to various fermented fish products ranging from 2 weeks in the case of some of the low-salt lactic fermented products to 5 years for some of the high salt sauces and pastes (16). the pH drops from 6.
viable numbers will probably decline quite rapidly. After capture. As in Norway. Because of the severity of the illness it causes. and failure to achieve a sufficient combination of inhibitory factors could lead to their growth or survival. The fish/salt/carbohydrate products depend on the combination of salt and pH/lactic acid for their safety. During fermentation. and consumed without cooking. Improper storage of fish prior to processing could result in the production of toxins capable of persisting through to the product. Their safety therefore is critically dependent on the quality of the raw materials used and the inhibitory effects of the fermentation process. under conditions inimical to growth and high ambient storage temperatures. fish products are the most common source of botulism. There is little in the way of documented information associating fermented fish products with foodborne illness. typically 30 °C. In Japan. and the most frequently implicated product is the fish-salt-carbohydrate fermented product Izushi. The bacterial pathogens most commonly associated with the living environment of fish. Several measures have been suggested for reducing the risk of botulism. however. V. 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. The Scandinavian fermented fish product rakefisk was responsible for 16 (84%) of the outbreaks of botulism recorded in Norway in the period 1961 90.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. parahaemolyticus has been reported growing down to a pH of 4. 6. The epidemiological evidence points to greater awareness of the importance of these factors since outbreaks have declined over recent years (27). the pH decreases below 5 and safe processing is recognised to depend on proper salt levels and temperature control until the pH drops. 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.5 Pathogens: Growth and Survival Many fermented fish products are stored at ambient temperatures.87. which accounted for 88% of outbreaks in the period 1952 . In the fish sauces and pastes.8 and at salt levels as high 10%. are Vibrio parahaemolyticus and Clostridium botulinum. but we are unaware of any reports of this happening. fish can potentially be contaminated with almost any foodborne pathogen as a result of improper handling and storage. outbreaks of 136 . although these limits apply when all other factors are optimal (26). The combinations of pH and salt in many fermented products are likely to preclude growth of the organism completely and. outbreaks of botulism are the only well documented instances of illness associated with fermented fish products.
. Saisithi P. botulinum and the production of toxin. Rakefisk and Herring in the Norwegian Tradition. Journal of Food Science. 6. Greenland and Alaska. Fermented Fish Products Study (Thailand) Phase 1. Dollar A. Infection with the trematode Opisthorchis viverrini is associated with cholangiocarcinoma. 3. viverrini were a strong risk indicator and that consumption of fermented fish products was an independent risk factor in the contraction of cholangiocarcinoma (31). they could act as an important vehicle of transmission. showed that inclusion of fermented fish in the Thai dish lab-pla improved inactivation of the metacercariae. and these are often used in the production of fermented fish products. salt content was found to have the greatest impact on the viability of larvae. affecting an estimated 40 million people.O.. An investigation of survival of the fish parasitic nematode Anisakis in marinated herring has given some indication of what these studies might find.. 1966. Lutefisk. Liston J. Puwastien P. walrus nose. Foodborne trematode infections are emerging as a major public health problem. The more widespread current use of non-traditional containers such as glass or plastic jars is thought to be one contributing factor (28). 31. A case-control study conducted in northeast Thailand showed that elevated levels of antibodies against O. while increasing the acetic acid content of the formulation had little or no effect (32). beaver tail and paw. Freshwater fish are the most important intermediate reservoir hosts of foodborne trematodes. 2. and salmon heads and eggs. 1990. although the absence of any source of fermentable carbohydrate means that the process is putrefactive. Microbiology and chemistry of fermented fish. 1989. probably reflecting greater awareness of the critical factors that determine the product’s safety (27). Raw materials subject to this process include seal flipper. Studies to determine the effect of traditional processing techniques on the infectivity of metacercariae was identified as a priority research area by a WHO study group (30). and warm weather can result in rapid growth of C.. Botulism has also been associated with a number of traditional foods of the peoples of northern Canada.. Baseline Study (1987-1988). References Riddervold A. though 27% survival was still recorded (33). They are packed into containers such as buckets or pits in the ground and left at ambient temperature to putrefy. Institute of Nutrition.6 1. Where these products are consumed without final cooking. With this particular organism. Many of these products are described as fermented. Dhanamitta S. Thailand. which is generally consumed raw (29). whale meat.M. One study with the trematode Haplorchis taichui.FERMENTED FISH botulism have become less frequent in recent years. Salting levels are often insufficiently low to prevent growth of C. Kasemsarn B. Yhoung-Aree J. Oslo. Mahidol University. 105-10. particularly in south-east Asia and the western Pacific regions (30). which will persist into the product. 137 . giving an alkaline fermentation. Novus Press. botulinum.
. 8. Namwong S. Fermented fish products of south east Asia. 1976. 2006. International Journal of Systematic and Evolutionary Microbiology.T. 70. Plenum Press.. Orejana F. from fermented shrimp paste in Thailand.. 57.H..W. 5. Rodtong S. Applied Microbiology. Adams M. 6338.V. Hull R.. Ono H.D. 1995. 57. Jae-Hun K. 25.. Microflora of four fermented fish sauces.. 1972. Process Biochemistry.H.. Olympia M. 16. Aslam Z. 185-92. Thawai C. SR5-3 isolated from fish sauce: purification and characterisation. Crisan E.T. Takada K.S. A halophilic serine proteinase from Halobacillus spp. Sands A. 353-63. 17-30.. Salinococcus jeotgali spp. Biotechnology and Biochemistry...A. 13.J. Tropical Science.. Catheptic activity of fish muscle. Yook H. 106-8.M. Fukuda H. Journal of Scientific and Industrial Research (India). Byun M. Yongsawatdigul J. Journal of Food Protection. 1975. Suntinanalert P. Nov. Phil.. 14. Ed. 1395-401.. Government of Australia. Tanasupawat S. 141-2. Dhamcharee B.. 124-30. Agents of proteolysis and its inhibition in patis (fish sauce). Liston J. 1957. Rodriguez-Valera F.. 15.. 61-73.. Production and characterisation of NaCl-activated proteinases from Virgibacillus spp. Hiraga K.R. isolated from jeotgal. Dymsza H. Ahn H.. 11. 138 . Effects of gamma radiation on sensory qualities. Kudo T. 1985. 63. Characterisation of starch-hydrolysing bacteria isolated from a fermented fish and rice food “burong isda” and its amylolytic enzyme. 16A. 17. Velankar N. Journal of Food Science and Technology. Protein hydrolysate from fish... 7. Oda K. 6. 11. Rattagool P..FISH AND SEAFOOD 4. 1986. 3649. 2007. 80. 12.. Reddi P...M. Nov. Journal of Food Science. 3. 43.. 934-9. New York. Yasir M. Constantinides S. Backhoff P. Sinsuwan S. Pakdeeto A. 2008. Lentibacillus kapialis spp. Tsunemi M.. Thongthai C. Itoh T.. Kyong-Haeng L. Bioscience.. Dong-Ho K. International Journal of Systematic and Evolutionary Microbiology. 643-7..K.. a traditional Korean fermented seafood.. Canberra.. A Concise Handbook of Indigenous Fermented Foods in the ASCA Countries. Halophiles in Thai fish sauce (Nam pla). 9.R. Lee S. in General and Applied Aspects of Halophilic Microorganisms. 37.R. Takano M.. microbiological and chemical properties of salted and fermented squid. Some chemical changes in fish silage. Cooke R. Saono S. Moonmangmee S. 29. Kaneko Y. 381-8. 1991. Lim J. Tanasupawat S.K. 2000. 2007. Chung Y.. Journal of Fermentation and Bioengineering.. 10. Im W. Journal of Food Technology. 1979. SK33 isolated from fish sauce fermentation..
2007. 28. a Thai low-salt fish product. a Thai low-salt fermented fish product. 28 (6). Ostergaard A. International Commission on Microbiological Specifications for Foods. Genotypic and phenotypic characterisation of garlic-fermenting lactic acid bacteria isolated from som-fak..H. in Clostridium botulinum: Ecology and Control in Foods. The influence of oxygen accessibility on the growth of yeast in fish/rice fermentation. 19. 20. 28.. Journal of Applied Microbiology. 27. 24.. Blackie. 103.FERMENTED FISH 18. 28. International Commission on Microbiological Specifications for Foods. 3. Eds. 1996. 23. 69-104. Rustad T. 138-44. Journal of Agricultural Science (Tokyo). in the Philippines.A. Ben Embarek P.D. Canada Communicable Disease Report. Two outbreaks of botulism associated with fermented salmon roe – British Columbia.. Fermentation and spoilage of som fak. 426-35.. 1998... 1999.. International Journal of Food Microbiology. Martin J. Caldo G. Vibrio parahaemolyticus.. 3-10.H. Wedell-Neergaard C. Yamprayoon J. Sakai H. Dodds K. Yeast-flora in red burong-isda a fermented fish food from the Philippines. Gram L. 22. Traditional fermented fish products with special reference to Thai products. 30. Characterisation of lactic acid bacteria isolated from a Thai low-salt fermented fish product and the role of garlic as substrate for fermentation. 21. 46. Isaac-Renton J.W. 38. Ed.. Gram L. Huss H. Marcel Dekker. Hauschild A.H. 105-12. 305-22. New York. Kozaki M.. London. Dodds K. Journal of Food Science and Technology. Marcel Dekker. Tropical Science. Erikson U. ASEAN Food Journal. Caldo G. Hazards from northern native foods. Hauschild A. Patrick D.. 2002. Journal of Agricultural Science (Tokyo). 1987. 1-4. Volume 5: Microbiological Specifications of Food Pathogens. 1992. Paludan-Müller C.H. in Microorganisms in Foods. 29. 27. 307-14. Valyasevi R.B.. Fung C. The fermented fish food.M. WHO Study Group... Epidemiology of human foodborne botulism.L.. burong-isda.A. World Health Organisation Technical Report Series. 2002.. Hauschild A.L. Benjakul S. Kozaki M. Control of foodborne trematode infections..W. Owens J. Visessanguan W.K. New York. 1992. 25. Effect of irradiation on properties and storage stability of Som-fug produced from bigeye snapper..H. Avurhi J. 1983. in Clostridium botulinum: Ecology and Control in Foods.W. Eds. 1995. Huss H. Saisithi P. 1815. Sakai H... Paludan-Müller C. 104-6. 1983..D. 1990... Riebroy S. Dawar M. 92. Tanaka M. 26. Food Chemistry. Gram L. 274-86. 219-29. Wainwright R.H. 139 .. Moody L..B. Huss H..
. 110-1.J. Gaithersburg. Southeast Asian Journal of Tropical Medicine and Public Health. Huss H. International Journal of Food Science & Technology.R. 661-70... 33. Thai traditional food preparation. Adams M. Uchida K.. 854-60.R.. 1995.. Kittiwatanachot P. Miwa M. Kikukawa H. northeast Thailand. Sripa B.. Srivatanakul P. 23.. 36. 1986. International Journal of Cancer.. Survival of Anisakis larvae in marinated herring fillets. Fermentation and Food Safety. Horst K. Roepstorff A. Progress in Industrial Microbiology. 32.7 Further Reading Adams M.. Fermented flesh foods. Todoroki T. Wongsawad C. Deerasamee S. 15998. Chuboon S. Hanai S. Survival of Haplorchis taichui metacercariae in lab-pla. Ruamsuk A.. Genetic and environmental determinants of risk for cholangiosarcoma via Opisthorchis viverrini in a densely infested area in Nakhon Phanom.. 140 . Nout M. 6. Bloemsma B.. 117. Nithikathkul C.R. Honjo S. Jedpiyawongse A. 2005...FISH AND SEAFOOD 31. Aspen Publishers Inc.H. 2005. 29.. 2001. Sriplung H...
FISH AND SHELLFISH TOXINS Jeffrey L. Another book dealing with the clinical effects of a variety of poisons and toxins. FCIC UNC Wilmington Center for Marine Science Marvin Moss Lane Wilmington NC 28409 United States of America 7. The manual on harmful marine algae and their products. a number of new toxins and toxin families have been identified. An emerging technology is the use of immunosensors (1 .1 Introduction Shellfish and finfish toxins continue to be a threat to public health and local fisheries around the world. In the last decade seafood monitoring has intensified and detection methods have continued to develop in selectivity and sensitivity. produced by the Intergovernmental Oceanographic Commission (IOC) of UNESCO (United Nations Educational. known and unknown.D. Scientific and Cultural Organisation) has been updated (7). Ph. the export of exotic seafood that may contain such toxins can lead to unexpected public health issues. physiology and detection of marine and freshwater toxins has also appeared (8).3) and some practical applications for detecting shellfish toxins have been reported (4. Increasingly sophisticated instrumentation (e. including the well known marine toxins 141 .C. and is often necessary in light of the complex toxin profiles that can occur in shellfish tissue.7. Although most of these antibody-based test kits have yet to receive international regulatory approval. While some of these toxins only occur in seafood from a restricted geographical region.. Wright. Nevertheless.M. Since the original chapter in this series was published (6). most notably the azaspiracids and the cyclic imines as well as some additional polyethers. Liquid Chromatography/ Mass Spectrometry/ Mass Spectrometry -LC/MS/MS) is employed to identify and confirm the presence of known toxins. It remains to be seen if this approach gains momentum in the future. 5). 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. and a second updated edition dealing with the pharmacology.g.. C. several scientific reports have described their efficacy and potential.
142 .2. with intoxications occurring in many countries even today (15). Saxitoxin (STX) and neosaxitoxin (neoSTX) are the parent members of the group and are the most toxic. neoSTX and Gonyautoxins (GTX) 1-4. Consequently. can become contaminated with these toxins. temperate. a great variety of shellfish from all latitudes. PSPs are watersoluble and heat stable so cooking has little or no effect on PSP toxicity. 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. 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. An account of this meeting detailing recommendations and scientific advice on marine biotoxins has been published (10).1 Origins and distribution The PSP toxins are produced by several dinoflagellate species belonging to the genera Alexandrium. The Ncarbamoyl sulfates are readily hydrolysed to STX.g.1). Around twenty PSP toxins are known. and sub-tropical waters and habitats (15). Not all the toxins found in shellfish extracts are microalgal products. A brief report on the status of rapid detection methods for phycotoxins in the European Union (EU) has appeared (11). and a tropical species Pyrodinium bahamense (16). The remarkable frequency of neurotoxins from marine dinoflagellates has been noted (13). and a review on the risk assessment of marine toxins has also been produced (14).FISH AND SEAFOOD has been produced (9). Many of these species are extremely common and are found throughout the world in a variety of cold. and another. Gymnodinium catenatum. All PSPs degrade rapidly in alkaline conditions. Saxitoxin Group instead of the Paralytic Shellfish Poisons). even crustaceans such as lobsters. 7. all of which contain a tetrahydropurine skeleton (Figure 7. some are the result of biotransformation processes by the filter-feeding host. 7. In addition.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. dealing with the detection methods and regulatory requirements for many marine toxins has been published (12).
2. and fingers within 5 . 7.30 minutes of consumption. Structure of saxitoxin and neosaxitoxin and their sulphated derivatives 7. PSPs bind to site 1 of sodium channels reversibly blocking sodium conductance in nerve and muscle membranes by binding to a specific receptor site located on the outside surface of the membrane close to the ion channel opening. tongue. Cooking does not destroy these compounds and although some of the toxins may be extracted into the cooking water. leading to paralysis within a few hours and in extreme cases death by asphyxiation due to respiratory paralysis.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. this is never sufficient to render the seafood safe to consume. Neurophysiological studies have shown that PSP toxins all possess the same mode of action though some derivatives such as STX and neoSTX are considerably more potent than the C-toxins (15). the less potent C toxins are readily hydrolysed by acid to the more potent GTX toxins. However. Human symptoms begin with tingling and numbness of the lips.1. 143 .2 Toxicology PSP toxins affect the central nervous system of mammals. and humans seem to be particularly susceptible.
While the chemical detection methods are most promising for the future.2. Since its discovery. Taiwan. 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 while simple and relatively inexpensive to operate. namely oxidative conversion to fluorescent derivatives after High Performance Liquid Chromatography (HPLC) analysis (post-column derivatisation). a criticism has been that such methods are time consuming. China. and islands of the South Pacific. Early post-column methods were somewhat time-consuming and required three separate HPLC runs to complete the analysis.8 µg/g) of shellfish or seafood tissue. 7. TTX most commonly occurs in many species of puffer fish. An antibody-based kit that can detect STX in shellfish is commercially available (RIDASCREEN®). Alternative HPLC methods have subsequently reduced this to a single HPLC run. and has also been evaluated by a number of EU laboratories. or conversion before HPLC separation (pre-column derivatisation) and these have recently been thoroughly reviewed (12. including Japan.3. 15.3 Detection methods The safe limit set by most countries for PSP toxins is 800 µg/kg (0.FISH AND SEAFOOD 7. One approach has been to take advantage of the ready conversion of these toxins to fluorescent derivatives.3 Tetrodotoxin Tetrodotoxin (TTX) possesses a unique chemical structure and was the first marine toxin to be chemically characterised.1 Origins and distribution Unlike many other marine toxins there is no temporal distribution of TTX episodes. Most intoxication events have occurred in South East Asia. There continues to be a significant effort applied to the development of alternative chemical and biochemical detection methods for these toxins. specifically 144 . a pre-column method has received Association of Analytical Communities (AOAC) approval (18). The mouse bioassay has been in place for decades and the method has been standardised. However it remains as the monitoring method of choice in many countries. 23. other derivatives have been reported. 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. 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. as well as other parts. and more standards of individual PSP toxins are required to validate the complete profile to toxins that could occur in contaminated shellfish tissue. 7. After some development and various trials. Thailand. and over ten analogs are known (22. and a second rapid dipstick kit (MIST Alert™) is available for the qualitative detection of positive and negative samples (19). 24) (Figure 7. Two approaches have been taken here.2). 17).
7. preventing depolarisation and propagation of action potentials in nerve cells.2. raising speculation that TTX is produced by a bacterial source.3 Detection methods TTX can be extracted from suspect tissue using dilute acid conditions. this has expanded to include bacteria within the Alteromonas and Pseudomonas genera. 25. nausea and vomiting. respiratory failure and coma. Severe poisoning causes paralysis and respiratory failure. TTX acts on the central nervous system by binding extracellularly to site 1 of voltage-gated sodium channels. and once again the mouse bioassay has been used to detect the presence of the toxin. However TTX has been found to occur in a remarkable variety of genetically unrelated species including gastropods. horseshoe crabs. The onset of symptoms usually occurs rapidly. An oral dose of 1 . 7. 23. and blocks diffusion of sodium through the sodium channel. and ingestion of larger amounts leads to cardiovascular effects including bradycardia.2 Toxicology Like the PSP toxins. All of the observed toxicity is secondary to the action potential blockade. gonads and roe (24). Chemical methods for TTX detection have also been investigated and are mainly based on its smooth conversion by heating in an alkaline solution to a detectable fluorescent (C9) base.FISH AND SHELLFISH TOXINS in the liver. Fig. Structure of tetrodotoxin 7. molluscs.2 mg of purified toxin can be lethal.3. the blue-ringed octopus. A review of TTX detection methods has been published (12. hypotension. it is served in speciality restaurants with qualified chefs to properly prepare the fish. where puffer fish of the genus Fugu are a delicacy. and in mild cases results in distal muscle weakness. 27). and more recently by an actinomycete Norcardiopsis dassonvillei (26). This was indeed confirmed and a number of marine Vibrios have been shown to be capable of TTX production (24. and even newts. Typically this has been accomplished using HPLC followed by post 145 . Since then. 26). In Japan.3.
both the toxin and the diatoms capable of producing it have been found to be distributed worldwide. Fig. a polar glutamate agonist. Structure of domoic acid and three naturally occurring photoisomers 7.1 Origins and distribution The cosmopolitan diatom Pseudonitzschia multiseries was identified as the culprit organism in the 1987 poisoning event (31). Immunoassay methods have also been investigated.3. but all of them are considerably less toxic than DA itself (29). but 146 . Surveys by a number of groups have identified over a dozen species of Pseudonitzschia capable of producing DA.FISH AND SEAFOOD column oxidation to the fluorescent derivatives. However this method has been criticised for being complex.4. These isomers arise as a result of isomerisation and location of the double bonds in the side chain of the molecule.3 for examples). Currently 12 isomers of DA have been reported (see Figure 7. and while promising. Since the original episode of human poisoning following consumption of DA-contaminated shellfish in Canada in 1987 (28). but it is known that such congeners are less toxic than DA itself (30).4 Amnesic Shellfish Poisoning (ASP) Toxins The parent compound of this group is domoic acid (DA). 7. need further development (24). 7. 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).
seizures. herbivorous fish become toxic after ingesting strains of Pseudonitzschia spp.FISH AND SHELLFISH TOXINS only three strains (P. Okinawa and more northerly parts of Japan. and Pseudonitzschia seriata) are considered to be high producers of DA (>10 pg/cell). has been reported as a medium-level producer of DA (33). including persistent short term memory deficits (36 . southern and western portions of Ireland. Pseudonitzschia australis. found in the Pacific around Vietnam. To a great extent. and in locations around New Zealand (32. The effects of DA may be augmented by the excitotoxic effects of glutamate and aspartate in the sample.10% of the administered dose. 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. which has been well documented in studies along the west coast of the US.. Penetration of the blood brain barrier is poor in healthy subjects. but any impairment of the blood brain barrier results in additional risk. Another factor is the movement of the toxin through the food chain. Recently. 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.4. Levels of DA in the other strains are generally <1 pg/cell. scallops. 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. 33). and review of the toxicologic pathology of DA has been published (36). as does impaired renal function which increases serum concentration and residence time of the toxin. multiseries. Pseudonitzschia and DA have been detected in many other maritime locations in the north Atlantic and the North Sea where it appears around the coast of the UK. Victims display a variety of symptoms including headache. 7. For example. Indeed. 147 . and clams. the Philippines. and is mainly distributed in the blood.2 Toxicology DA is an acidic amino acid and potent neurotoxin unaffected by cooking or steaming. Four deaths were attributed to the Canadian incident and years later a few seriously affected individuals still display chronic neurological deficits. disorientation and coma. another diatom species Nitzschia navisvaringica. and this has led to ASP toxicity in birds. 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. nausea. cormorants and pelicans (34). the Mediterranean.39). and sea animals such as sea lions and even whales (35). The oral absorption of DA is around 5 .
Dinophysis spp. 7. and dinophysistoxin-2 (DTX-2) (Figure 7. the term “DSP toxins” will only be used to refer to these three polyether compounds and any derivative based on them. DTX-2 has been found to be extremely widespread throughout many European maritime countries and is often the major DSP toxin present. Since then. and an immunosensor method has also been reported (4). Pseudonitzschia fraudulenta and Pseudonitzschia delicatissima were found to be less effective. but just over 15 years ago a third toxin. DTX-2 was discovered in contaminated Irish shellfish (46). the heterotrophic dinoflagellates Dinophysis spp.3 Detection methods DA is water soluble and can be extracted from shellfish tissue by the acidic PSP extraction method although this may result in some decomposition. Provided the concentrations in shellfish are high enough (>50 µg/g shellfish tissue). produce a variety of other 148 . Since their first characterisation it was believed that the only causative agents of this group were OA and DTX-1 (45). 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). 49). An EnzymeLinked ImmunoSorbent Assay (ELISA) kit (Biosense) for DA has been approved by the AOAC. are the more potent source of toxins – often very low cells/ml of seawater can result in human poisoning. though this method is most commonly used to analyse plankton and seawater (42). The benthic Prorocentrum spp. although in the latter study the probes for Pseudonitzschia pungens. and consequently were often overlooked as the real source of the toxins.. are amenable to laboratory culture and consequently are used in many experiments involving studies of DSP toxins. Alternatively.30 ng/g) with a rapid clean-up step using a strong anion exchange resin. 41). 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)..4). California. and this is typically the method of choice. However. as can LC/MS.FISH AND SEAFOOD 7. Detection levels can be further lowered by conversion of DA to the fluorescent fluorenylmethoxycarbonyl (FMOC) derivative. The sensitivity of the LC/UV (ultra violet) method (detection limit ~ 1 µg/g) can be improved (20 . 48. DSP toxins are produced by dinoflagellates of the genera Prorocentrum and Dinophysis (47. shellfish tissue can be efficiently extracted with aqueous methanol (1:1). In the context of this chapter.4. australis. To further complicate matters. Of additional interest is the use of molecular probes originally developed to detect the presence of cells of potentially toxic Pseudonitzschia spp. Fortunately DA is conveniently detected by a variety of LC-linked analytical methods and this is the recommended approach for DA monitoring in EU countries. and these methods have been reviewed (40. The probes have been employed in trial studies in New Zealand (44) and Scotland (43) to detect P. dinophysistoxin-1 (DTX-1). in Monterey Bay.
7. and include Dinophysis acuminata. but do not have the same mechanism of action as DSP toxins nor induce the same symptoms. Lately these compounds. 49). Dinophysis fortii. which include the pectenotoxins and the yessotoxins. and DTX-6) all of which are rapidly converted to OA when the plankton cells are ruptured (48). Dinophysis acuta. 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. 149 .4. Prorocentrum maculosum. This was probably not the first DSP episode. In comparison. and Prorocentrum belizeanum.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. DSP toxins produced by benthic Prorocentrum spp. are recognised as DSP toxin producers. Structure of okadaic acid and the two related DSP toxins 7. DTX-5. and anecdotal evidence now suggests that several seafood poisoning episodes during the 1960s and 1970s in the Netherlands.FISH AND SHELLFISH TOXINS lipophilic compounds that are toxic in the mouse bioassay. 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. While these esters are not toxic in vitro. Dinophysis caudata. Fig. analysis of Prorocentrum cultures have uncovered an additional series of diol esters of OA and sulphated diesters (called DTX-4. The parent acids are the only DSP congeners to be found in shellfish tissue. such as Prorocentrum lima.. they can be converted to the parent toxins by nonspecific esterases or lipases either in the producing organism or in the host shellfish. are also recognised as a threat to human health. Today over ten Dinophysis spp. Norway and Japan were likely due to DSP toxins produced by a Dinophysis spp. In this case Dinophysis was suspected as the culprit organism.5. and Dinophysis norvegica among others (47.
DSP episodes have been reported from Japan. and while this provides critical seafood safety information. 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). it does not provide information on the DSP toxin profile. At the first Meeting of the EU National Reference Laboratories on Marine Biotoxins and Analytical Methods and Toxicity Criteria in 1996. 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.5. Preparation of a fluorescent 9-anthryldiazomethane (ADAM) derivative followed by HPLC offers reasonable sensitivity. DSP episodes have been reported worldwide and are included in many monitoring programs (47).FISH AND SEAFOOD Perhaps as a consequence of these multiple sources of DSP toxins. This regulatory level includes the possibility of LSPs in the DSP fraction being tested. Due to the lipophilic nature of the toxins. and DTX-2. vomiting. some careful clean-up steps are required to avoid such matrix interferences and to simplify data interpretation (48). 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). 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. various European countries. A regulatory level of 80 . and a detection limit of 100 µg/kg has been reported (12). most progress in DSP toxin detection has been using new or improved LC/MS methods and a detection limit of 10 µg/kg has been reported. All the DSP toxins have around a thousandfold greater affinity for PP2A than PP1.160 µg OA equivalents/kg of whole shellfish tissue has been implemented in some countries.3 days. 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. nausea.5. The onset of symptoms can occur within 30 minutes of consuming contaminated shellfish and generally last about 2 . and abdominal pain. In recent years. which can lead to an underestimation of the total DSP content in a sample.2 Toxicology The DSP toxins are potent inhibitors of the serine/threonine phosphatases PP1 and PP2A. even with the minimal of clean up steps (48). DSP-Check is a commercially 150 .3 Detection methods Once again the mouse or rat bioassay is relied upon to detect DSP toxins in contaminated seafood. Immunoassays have been developed for OA detection but an inherent problem of these methods has been the low cross reactivity with DTX-1. New Zealand and Chile. Human symptoms include diarrhoea. Since the late 1970s. 7. 7. but the method is complex and matrix effects as well as interferences with other lipophilic compounds such as the pectenotoxins compromise the method.
they are toxic in the mouse bioassay and can result in false positives and hence are included here for completeness. 7. Within this group. 7. and the latter group are usually found to be responsible for any reported shellfish toxicity (59). fluorometric protein phosphatase inhibition assays have been found to perform better than colorimetric assays. Indeed. Some doubt remains as to the significance of these compounds as marine or shellfish toxins with respect to human health since the PTXs for example are co-extracted with the DSP toxins. 58). for many years these compounds were included in the DSP toxin complex. Structure of the principal pectenotoxins and one of the seco-acid derivatives 151 .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). Some representative examples of both groups of toxins are shown in Figures 7. In yet another approach.5 and 7. Fig. over 20 PTXs are known (56) and more than 30 YTXs have been reported (57. with a claimed detection limit of 20 µg/kg. 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. with good agreement with the mouse bioassay and LC–based methods (54. However. 55). As a result these compounds are now being referred to as lipophilic shellfish toxins (LSTs) (49). and another rapid antibodybased test kit for DSP toxins has also been reported (53).FISH AND SHELLFISH TOXINS available ELISA test kit that can detect both OA and DTX-1 with comparable sensitivity. 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.5.6.
As is typical for many algal species. New Zealand and South America (49.FISH AND SEAFOOD Fig. YTX and homo-YTX 7. eastern Canada.6. 58). which may co-occur in blooms with other organisms.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). 57. but nevertheless the toxins are extremely widespread and have been found in various shellfish in Norway and Scandinavia.6. Structure of the two parent yessotoxins. 7. many parts of Europe and the Mediterranean.2 Toxicology PTXs show varied toxicity in the mouse bioassay (59). the toxin content and profile may vary among different strains. 7. Oxidation of the methyl group at C-43 to a carboxyl function as in PTX-6 significantly reduces toxicity (500 µg/kg body weight) as does hydrolysis to PTX-2 seco acid (>5000 µg/kg body weight) a common metabolite found in shellfish extracts. Japan. 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). While 152 .6. The sources of YTXs have now been identified as the dinoflagellates Protoceratium reticulatum and Lingulodinium polyedrum.
but various congeners are acutely toxic in the mouse bioassay following IP injection. The YTXs are retained in the aqueous methanol fraction while the PTX and DSP toxins are concentrated in a dichloromethane fraction. chemical analysis is best carried out by LC/MS and several approaches have been described (57. For many years it was believed that 153 . but less so following oral dosing (61). which aids detection by LC/UV.3 Detection methods Typically these toxins are co-extracted with DSP toxins following acetone or aqueous methanol extraction of the shellfish tissue. 64). 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. 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. though its cell effects may be linked to modulation of calcium channels (62). which is further compounded by differences in the experimental design. and this is believed to be due to hydrolysis of PTX-2 to the seco acid derivative. There are no clear cases of human illness resulting from ingestion of PTXs (59). some of which are found as glycosides and sulfates. The crude extract may be used directly in the mouse bioassay. 7. Although there is no evidence that yessotoxins are responsible for human illness. Some of the PTXs are potent cytotoxins in vitro (60). There are two skeletal types known as PbTx-1 (or brevetoxin A) and PbTx-2 (brevetoxin B) (Figure 7. Because of the number and complexity of the YTX group. The EU has recommended an extraction and solvent partition process to separate the polar YTX compounds from the more lipophilic PTX and DSP compounds. 58). a regulatory level of 1 mg/kg shellfish tissue has been set. However. 7.7 Neurotoxin Shellfish Poisoning (NSP) Toxins NSP episodes are caused by a family of fused polyether compounds known as the brevetoxins (63.7). Estimates of acute toxicity of the parent compound following IP injection vary considerably.FISH AND SHELLFISH TOXINS demonstrating considerable toxicity following IP injection. it may be compromised by other biological materials in the matrix. but a fluorescent derivative has been prepared for others. The structures and chemical data including the molecular weight of each member have been collated (56). Acetone extracts of shellfish tissue are dried and re-suspended in 60% aqueous methanol before partitioning against dichloromethane. Many YTX toxins contain a conjugated diene system. Their toxicology and mechanism of action remain uncertain.6. While LC/UV detection is possible for PTXs. the PTXs show considerably less toxicity when administered orally.
Fibrocapsa japonica. and off the coast of New Zealand (65).7. 154 . was chemically characterised as PbTx-2 (67).7. namely Chatonella marina. oysters. PbTx-1 and PbTx-2. Fig. brevetoxins are found in mussels.S. whelks. and one of the brevetoxin metabolites BTX-B2 7. 7. and recently the major toxin associated with an ichthyotoxic unialgal bloom of Chatonella cf. Structure of the two parent brevetoxins. the same toxins have been reported to be produced by three genera of Raphidophytes. Interestingly. which are found in Japanese waters (66). and cockles (65. They have also been associated with fish kills along the eastern mid-Atlantic coast of the US. 66). verruculosa off the mid Atlantic coast of the U.FISH AND SEAFOOD NSP toxins only occurred in the Gulf of Mexico and along the western coast of Florida. These are biotransformation products of principally PbTx-2 by addition of certain amino acids to the conjugated aldehyde function (66). A complicating factor both in terms of establishing the profile and toxicity of brevetoxin-contaminated shellfish has been the discovery of the so-called “brevetoxin metabolites”. but in 1992/1993 they were identified as the culprits in a human poisoning event in New Zealand (65. 66). Many of these species are associated with massive fish kills.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. In the Gulf of Mexico. and Heretosigma akashiwo.
The mouse bioassay is used as the primary detection method. and in severe cases seizure. 7.8 Ciguatoxins Known for centuries and originally referred to as ciguatera by Spanish explorers in the Indo-Pacific ocean and Caribbean.3 Detection methods Three basic approaches for the detection of NSP toxins have been recently reviewed (71).7.FISH AND SHELLFISH TOXINS although recently it has been shown that herbivorous fish can store brevetoxins in their flesh (68). considerable progress has been made in the development of LC-MS methods to routinely monitor for brevetoxins in shellfish. but this method is now used routinely in New Zealand and the US to monitor for the presence of brevetoxins (68). the competitive ELISA method correlated most closely with the mouse bioassay results. shellfish homogenates. A second generation competitive ELISA assay has been developed that has successfully quantified brevetoxins in a variety of biological and environmental matrices including seawater. which can result in a cascade of associated events in a cell (66). Ingestion of brevetoxins results in a variety of symptoms including parathesia. The human health implications following exposure to NSPs have been reviewed (69). and mammalian body fluids without pretreatment or dilution (72). beginning with herbivores that feed on benthic dinoflagellates associated with reefs and which are 155 . vertigo.2 Toxicology Brevetoxins bind to site 5 of voltage-gated sodium channels. While the mouse bioassay continues to be used. nausea. The mean time to onset of symptoms is around 3 hours. Outbreaks of ciguatera poisoning are confined to tropical and sub-tropical regions of the ocean. Exposure to brevetoxins can also occur through inhalation of brevetoxin aerosols resulting in irritation of the upper respiratory tract (65. 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. Ciguatera toxins (CTXs) are extremely potent and occur in finfish rather than shellfish. with a mean duration of symptoms of around 1 day. pre-treatment or clean-up of the shellfish sample is essential in order to obtain reproducible results. this family of toxins has proven to be a challenge both to identify and monitor. malaise. 69). 7. In a comprehensive multi laboratory evaluation of five different detection methods as possible replacements for the mouse bioassay. 66. The brevetoxin metabolites are reported to be less toxic than the parent compound (70).7. with hot spots in the coral reef areas of the Caribbean and French Polynesia. 7. In this application. and it has also been used in studies to quantify occupational and recreational exposure to aerosolised brevetoxin exposure (73). and diarrhoea.
A detailed comprehensive review of ciguatoxins has recently been published (74). The diversity of benthic phytoplankton that exist on reefs further complicates the picture. CTXs are large. and only a few structures have been identified. although the dinoflagellate Gambierdiscus toxicus is considered to be the major source of these toxins. and other genera of neighbouring reef dinoflagellates can further contribute to an extremely complex profile of toxins in finfish. Such microalgae contribute to the primary production of tropical and sub-tropical reefs and live in epiphytic association with bushy red.8.1 Origins and distribution It is recognised that several species of benthic dinoflagellates can produce CTXs and the gambiertoxins.FISH AND SEAFOOD transferred through the food chain to the larger carnivores.8. complex. Culprit species producing CTXs may simultaneously produce unrelated toxins such as maitotoxin.8). green and brown seaweeds and also occur in sediments and coral debris. but more are suspected to exist. Indeed even the chemistry of CTXs from the Pacific differs subtly from those found in the Caribbean (Figure 7. Distribution through the food chain begins with 156 . lipid soluble polyethers. most likely a consequence of biotransformation of the parent toxins as they pass up the food chain. Fig. Structure of the Pacific variant of ciguatera toxin (P-CTX-1) and the Caribbean variant (C-CTX-1) 7. 7. a group of closely related polyethers that are considered the source of ciguatoxins. These include species belonging to the genera Ostreopsis and Coolia.
the absence of a strong chromophore means that simple detection methods based on UV or fluorescence-based detection methods are ineffective. Other symptoms include a metallic taste. pruritus. sharks. which can return following the consumption of alcohol and certain foods such as oily fish (74. there are neither standards nor an official testing program. though the mouse bioassay continues to be relied upon as the most common method of detecting CTXs. 78).8. weeks or even months (74 .FISH AND SHELLFISH TOXINS grazing by herbivorous fish including the sturgeon fish and parrotfish. and it is estimated that ingestion of as little as 0. Like the brevetoxins. 7.76). groupers. and temperature sensation reversal. Nevertheless CTX poisoning continues to be a threat to local and visiting populations. but fish toxicity can vary widely depending on where the fish are caught and how long they have been feeding in that area. 75). 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. Differences in potency among the CTXs correlate directly with their binding affinity for the sodium channel. Another symptom that can develop over time is extreme fatigue. Even when suspect samples are available. they bind to voltage-dependant sodium channels in the cell membrane causing them to open. The pharmacological effects following CTX exposure are numerous and include severe gastrointestinal distress and neurological symptoms.1 µg of ciguatoxin can result in human illness (74.2 Toxicology Ciguatoxins (and maitotoxin which is often co-produced with CTXs) are extremely poisonous. which can be confused with chronic fatigue syndrome. and inducing membrane depolarisation resulting in prolonged symptoms indicative of damage to nerve tissue.3 Detection methods As a consequence of their potency.8. 7. and there are suggestions that such incidents are increasing and spreading worldwide (74). only trace amounts of CTXs can cause illness following consumption of contaminated fish. where cold objects feel hot and vice versa. and although the gastronomical problems may dissipate within a day or so. barracudas. although not all CTXs are equipotent. Globally. all of which can be linked to the effect of CTX on excitable membranes. Patients have reported symptoms lasting weeks and even months. arthralgia. A good knowledge and careful observation of the symptoms in the bioassay (Minimal 157 . which in turn are consumed by carnivores such as snappers. 75. These include paresthesias (numbness and tingling of the extremities). The onset of symptoms can occur rapidly within 24 hours. Local island populations attempt to avoid poisoning by catching only smaller fish. and moray eels. and in fact CTX poisoning is often only identified after consumption of contaminated fish. the neurological symptoms can persist for days. myalgia. and the sensation of loose teeth (77).
Usually the additional AZA derivatives found in shellfish tissue are hydroxylated derivatives (e. and following synthesis of the molecule some time later. and the separation of these polyethers from the lipid-soluble fraction of fish provides an additional challenge. 84). has been developed. although specificity and sensitivity remain major hurdles (74) and the presence of DSP toxins affects the test (80). Consequently. specific for CTX activity.30%) and AZA3 (5 . another in vitro sodium channel-based assay. A sodium channel-specific assay using mouse neuroblastoma cells has been described although this approach has still to be developed in a routine.5) have been fully characterised by Nuclear Magnetic Resonance (NMR) and MS analysis. high-throughput screen (74).20%) present in lesser amounts (Figure 7. and only a few structures (AZA 1 . Immunoassays may be an alternative detection method (79). only a few laboratories are able to undertake such analyses. with AZA2 (10 . The original structure proposed revealed an entirely new class of shellfish toxin. and it is likely that most of these compounds are biotransformation products formed in the shellfish tissue. following consumption of contaminated mussels Mytelius edulis (82).9 Azaspiracids Azaspiracid (AZA1) was originally identified in 1995 as the causative agent for a poisoning episode in Ireland. and the corrected structure was published in 2004 (86) (Figure 7.9). which features characteristic fragment ions corresponding to sequential losses of three molecules of water. 7. which are based on an LC-MS/MS approach. 158 . but this only has application in a clinical setting (81). minor revisions of a few stereocenters were made. Since the original discovery and identification of the toxin. followed by clean-up on silica and aminopropyl cartridges. and another dealing with the chemistry and detection methods has been published (85).25 µg/kg) can provide useful clues as to the toxins present but more specific and informative assays are required.85). Recent reviews of this new group of toxins have appeared (83. Usually the major AZA derivatives found in shellfish are AZA1 (50 . In this way Caribbean and Pacific CTX-1 compounds can be distinguished (74). almost 20 additional derivatives have been reported (83 . Recently. Chemical detection methods are severely compromised by the lack of standards. Acetone extracts of fish tissue are subjected to solvent partitioning.FISH AND SEAFOOD Lethal Dose (MLD) 0. The majority of these have been proposed based on LC/MS analytical data. This is then subjected to LCMS/MS analysis.g. at C-3 and C-23).80%).9).
and most recently in a crustacean (Cancer pagurus). 7. Structure of the three principal azaspiracids found in shellfish 7. In fact the routes of trophic transfer of AZAs through the food chain ending in contaminated shellfish remain unknown. Donax spp. which are ingested by P. Following the original episode. While this result is not necessarily in debate. 88. AZA toxins do 159 .2 Toxicology Consumption of AZA contaminated shellfish can result in acute symptoms that include nausea.). Portugal.9. However. oysters (Crassostrea gigas. Ostrea edulis).3 days. 90). and variable field results have led to suggestions that the actual producing organisms are Dinophysis spp.FISH AND SHELLFISH TOXINS Fig. 84. and to date this has been the only source from which these toxins have been found at toxic levels. Scandinavia. diarrhoea and stomach cramps. The bivalves affected include mussels (Mytilus edulis. and some time after the original episode in Ireland. and off the coast of Morocco (83. cassipes produces AZAs in culture. vomiting. These symptoms are not unlike those experienced following the consumption of DSP toxins. Mytilus galloprovincialis). 89). these molluscs were never considered to be the biogenetic source. cockles (Cerastoderma edule). which can persist for 2 . cassipes (83). it was reported that the heterotrophic dinoflagellate Protoperidinium cassipes was the planktonic source of the toxins (87).1 Origins and distribution The azaspiracids were first isolated from contaminated shellfish. and to date no long-term effects have been recorded (83. but current data indicates that unlike the DSP compounds. AZA occurrence has been reported by several European countries including France. There has been considerable debate over the source of azaspiracids. it has been impossible to establish that P. clams (Tapes philipinarum. scallops (Pecten maximus). Ensis siliqua. 7.9.9. as in other cases of shellfish toxins.
PSP. 91. though the possibility exists that they may inhibit other phosphatases. and closely related ptierotoxins (Figures 7.12).10 – 7. it is believed that the hydroxylated derivatives AZA4 (470 mg/kg) and AZA5 (<1000 mg/kg) are considerably less toxic. Due to their relatively recent discovery and limited knowledge of their toxicological properties and impact on human 160 . This common cyclic imine feature is believed to be a common pharmacophore of this group of toxins which comprises the gymnodimines. Once again. the pinnatoxins. However. 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. parenchymal cell pyknosis in the pancreas.9. Importantly these pathological outcomes were reported as being different from those caused by DSP. 85). 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. Injection of a lethal dose of AZA1 in mice (>150 µg/kg) caused vacuole formation and fatty acid accumulation in hepatocytes. and ASP. and erosion and bleeding in the stomach. it is suggested that the order of toxicity is AZA2 (110 µg/kg) >AZA3 (140 µg/kg) >AZA (150 µg/kg). and necrosis of epithelial cells on the microvilli (91). The mouse bioassay has been used to monitor shellfish extracts. Key fragment ions are useful in identifying the various AZA compounds that can occur in shellfish tissue (83. although autopsies conducted after 4 and 8 hours revealed various gastrointestinal (GI) abnormalities including accumulation of fluid in the ileum.FISH AND SEAFOOD not inhibit the phosphatases PP1 and PP2. the spirolides. The limited amounts of AZA toxins available continue to hinder a complete understanding of their mechanism of action. Using the same approach.10 Cyclic imines These toxins are represented by several distinct structural types. which are particularly amenable to detection by positive ion electrospray mass spectrometry. 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). 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. 7. AZAs also display cytotoxicity towards various mammalian cell lines (83. 7. but all contain a spirocyclic or cyclic imine function within their structure. Based on IP minimum lethal dose data. abnormalities in the thymus and spleen.3 Detection methods The regulatory level for AZAs in shellfish has been set by the EU at 160 µg AZA equivalents/kg shellfish tissue. but unlike DSP monitoring methods which use only the hepatopancreas of the shellfish for extraction. 92).
1. 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. cockles.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. Structure of the cyclic imine gymnodimine 7. 95). gymnodimine B and C were isolated from the dinoflagellate Karenia selliformis (formerly Gymnodinium selliformis). 7. Some time later. a species closely related to Gymnodinium mikimoitoi was the planktonic source of the toxin.10.1 Gymnodimines The observation of neurotoxic symptoms in the mouse bioassay of lipophilic extracts of dredge oysters (Tiostrea chilensis) collected from around the South Island of New Zealand. surf clam and New Zealand abalone. scallops.10. The structure is novel in that it contains a spirocylic imine group and a single tetrahydrofuran ring embedded within a macrocyclic structure. led to the identification of a new marine toxin with an unprecedented structure named gymnodimine (96) (Figure 7. two additional congeners. they are sometimes referred to as emerging toxins of uncertain human health concern. it was proposed that a dinoflagellate identified as Gymnodinium spp. Fig. At the time of the initial observation of gymnodimine.FISH AND SHELLFISH TOXINS health. 161 .. selliformis (97).10. The gymnodimines are the smallest members of the cyclic imine group with a molecular weight range of 500 . but they are included here for completeness. Reviews of the current status of the cyclic imine group were published by Molgo and Cembella (94. 95). In only two cases (gymnodimines and spirolides) has a planktonic source been identified. Pacific oyster. 7.10).520 Da (94. Subsequent taxonomic studies revised this again to a new dinoflagellate K.
100). the first step in the process is typically an initial clean-up step using a short C18 cartridge and elution with water containing increasing amounts of methanol. 95). extracts exhibited the fast acting symptoms that became associated with cyclic imines following IP injection. Whether isolated from dinoflagellate cells directly or from shellfish tissue. There is a rapid onset of symptoms including respiratory distress.3 Detection methods The characteristic “fast-acting. cyclic imines are readily observed by electrospray mass spectrometry in the positive ion mode. Most cyclic imines lack an extended chromophore. edulis) and scallops (Placopectin magellanicus) from eastern Canada that displayed abnormal symptoms in the mouse bioassay. Oral administration of the toxin to mice by gavage resulted in about an 8. A later study using purified toxin established an LD50 dose (Lethal Dose.10. and lack of responsiveness to stimuli. all or nothing” mouse bioassay response to gymnodimines and cyclic imines in general continues to be a useful monitoring approach.2 Toxicology A review of the toxicology of all cyclic imines has been published (98).10. 50%) of 96 µg/kg. together with a lack of evidence of human intoxication. finishing with 100% methanol. Since their original discovery which resulted in the characterisation of spirolides A-F (99.1.FISH AND SEAFOOD 7.2 Spirolides The spirolides comprise the largest group of cyclic imine compounds.to10-fold reduction of toxicity (LD50 600 . They were originally discovered in extracts of mussels (M. Based on this diminished oral toxicity. The time lapse between ingestion and death is between 5 and 15 minutes.11). The structure of the spirolides bears a strong resemblance to both gymnodimine and 162 . 7. and this is the detection method of choice. it has been stated that regulation against gymnodimine is not required (98). almost a dozen derivatives in total have been identified to date (94. This led to the suggestion that gymnodimine acts by blocking nicotinic acetylcholine receptors at the neuromuscular junction. However. 95) (Figure 7. and so spectroscopic detection by UV methods is not favoured.900 µg/kg). abdominal breathing.1. The original observation of toxicity in New Zealand followed from IP injection of lipophilic extracts in the mouse bioassay. Fractions are then dried and re-suspended in methanol or acetonitrile in preparation for LC/MS analysis (94. Reduction of the imine group eradicated the toxicity. and there was a further ten-fold reduction in toxicity when the toxin was supplied with food. paralysis of the hind quarters and eventually complete immobility.10. and short-acting cholinesterase inhibitors such as neostigmine or physostigmine protected mice against the toxic effects of gymnodimine. 7. In particular.
in fjords in Denmark and Norway. The LD50 values vary considerably depending upon whether the compound is administered by IP injection (5 . those spirolides possessing the vicinal dimethyl grouping (see Figure 7. Structure of four spirolide toxins A-D 7.2 Toxicology The spirolides are the most studied group of the cyclic imines.1 Origins and distribution At the time of the original discovery of the spirolides in shellfish from the coastal waters of eastern Nova Scotia. 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.10. 7. the gulf of Maine. fasted mice). Fig. by gavage (87 . or by oral ingestion (381 .11.2. the Bay of Biscay and most recently from Mediterranean shellfish and plankton in the Adriatic (102).707 µg/kg. fasted mice). Since then the spirolides have been found to be globally distributed and are found in shellfish or plankton fractions along eastern Canada. 7. 95). R1 = CH3) of the cyclic imine ring are the most toxic of the group (98). This proved to be the case and eventually the dinoflagellate Alexandrium ostenfeldii was identified as the culprit organism (101).8 µg/kg). They have also been reported in the southern hemisphere in Chilean mussels and in New Zealand shellfish and plankton (94. Interestingly. the western coast of Scotland. these are the toxins that are also most resistant to acid hydrolysis of the imine function 163 . southern Ireland.10.FISH AND SHELLFISH TOXINS the pinnatoxins (vide infra).2. While oral toxicity is considerably less than by IP injection. All data reported here are for desmethyl spirolide C. their localisation in the digestive gland of mussels and scallops led to the suspicion that plankton were the real source of the toxins.166 µg/kg.11.
which includes two diastereomers. and in fact these compounds share a number of other structural features.12. 95. 102). Once again this relies on the sensitivity of these cyclic imines to detection by positive ion electrospray mass spectrometry.FISH AND SEAFOOD (100). While the mechanism of action remains to be clearly defined. experiments have shown upregulation of muscarinic and nicotinic receptors in mice following dosing with spirolide C.12). The pteriatoxins were isolated from a different mollusc but bear a strong structural resemblance to the pinnatoxins (Figure 7. 7. 7. A number of methods following this procedure have been described (94.B contain a spiro imine moiety similar to that found in the spirolides.3 Pinnatoxins and pteriatoxins Pinnatoxins A . Structure of the pinnatoxin group and the closely related pteriatoxins 164 . However. 7.10.10. The compounds are zwitterionic and therefore somewhat more polar than the spirolides. To date. the mouse bioassay can be used to detect these fast acting toxins. Fig. four members of the group have been identified – two of them being the C-34 diastereomers.2.3 Detection methods As discussed for gymnodimine. Three members of this group have been reported. 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. consistent with modulation of acetyl cholinergic receptors (103).
1 Origins and distribution Shellfish of the genus Pinna.3. while pinnatoxin D was essentially inactive. has come the realisation that a “toxin profile” can be a complex mixture of many different classes of toxins. There is strong circumstantial evidence to suspect that these toxins. With increasing knowledge of the chemistry of toxins. Despite this progress.2 Toxicology There is little information describing the toxicology of these compounds (98). new toxin groups such as the azaspiracids and cyclic imines have also been reported. Coupled with this. and more advanced means of detection. This is generally a result of increased monitoring and continually improving detection technologies and methods.FISH AND SHELLFISH TOXINS 7.3 Detection methods No formal detection methods for these compounds have been reported.most likely a dinoflagellate.10. 7.10. This prompted a chemical investigation of Pinna muricata that ultimately led to the identification of the pinnatoxins in 1995 (104). The LD99 dose for the pteriatoxins is reported to be in the range (8 . However. which occupy tropical waters around Okinawa and the south east coast of China. arise from an algal source .180 µg/kg).100 µg/kg). like the gymnodimines and spirolides. While this is recognised as a real possibility for a wide range of toxins.3. Only the acute toxicity in mice following IP injection has been reported. 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. 7. but this remains to be conclusively established (106). 99%) values for pinnatoxins A-C are in the range (22 . there is no compelling evidence to suggest that they have resulted in human toxicity (98). once again reflecting improvements in purification and spectroscopic methods permitting structural identification of new and complex structures. The LD99 (Lethal Dose. 7. which in many cases are now found to be more widespread than previously believed. but the mouse bioassay data and LC-MS can be used to monitor suspect seafood for these compounds. have long been suspected of causing human illness after ingestion.3.10. and later the closely related pteriatoxins were isolated from the digestive glands of another Okinawan bivalve Pteria penguin (105). and in some cases it is known that one toxin can potentiate or augment the toxicity of the other. to date action or 165 .11 Conclusion There has been considerable progress in the detection of known marine toxins.
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Botana L.. 23. 3. Poli M.G...P. Bourdelais A. Ed. Ciguatera fish poisoning in industrial ship crew members: a retrospective study in a seaport general practice in Trinidad and Tobago. Baden D..P.G... 50.D. Ed. Bottein M-Y. Baden D.. 2008.. 2000.A. Granholm A.G. Bean J. Pierce R. like ciguatoxins. 67. 68. Flewelling L. are potent ichthyotoxic neurotoxins that accumulate in fish. Naar J.. 2004.E.C. Food and Agriculture Organisation of the United Nations.B. 187-94. Henry M. Weisman R. Toxicon.G. 70.M. Gandini C. Ed. 179-85.. Boca Raton. Ayyar R.A.. Boca Raton.E. Literature review of Florida red tide: implications for human health. 456-79. Ed. CRC Press. Wang Z.. Ramsdell J. New fish-killing alga in coastal Delaware produces neurotoxins. 74. Johnson D. 75. Fleming L. in Seafood and Freshwater Toxins. 220-6.A.... Watkins S. 66.. and detection. Fernandez M. Backer L.G..S. Steidinger K. 110... 2004. Poon-King T. Zaias J. Mild ciguatera poisoning: case reports with neurophysiological evaluations.J. Pierce R. Cheng Y. Baden D. Fleming L. 110. 2008.. CRC Press. Tomas C. Ciguatera fish poisoning: treatment prevention and management. Prockop L. in Seafood and Freshwater Toxins.....E. CRC Press.D... The molecular and integrative basis to brevetoxin toxicity...... Muscle Nerve. 69. Toxicon. Backer L. West Indian Medical Journal. Aerosolised red tide toxins (Brevetoxins) and asthma. Naar J..P. Abraham W.. Intrinsic potency for synthetically prepared brevetoxin cysteine metabolites BTX-B2 and desoxy-BTX-B2.. Marine Drugs.. Buonocore M. Clark R. Benson J. Lenzi A. 6. Chest. Dickey R.M. Baden D.... Manzo L. Tomas C. Environmental Health Perspectives..W. Dickey R. Bossart G. 2002. Kubanek J. 551-8. Kubanek J.M... 76..C. Food and Agriculture Organisation..E. 71. Brevetoxins. Backer L. Dechraoui M-Y.... Bourdelais A.A. Wanner A.. 72. Jacocks H.. Ramsdell J.. toxicology.... shellfish and mammalian body fluid. Landsberg J. 2008. Pierce R. Squicciarini D. Harmful Algae. 1598-603.. Chen A. Ciguatera toxins: Chemistry.. 2008. Gannon D..N. Boca Raton. 2004. 131. Rome. Flewelling L. Kirkpatrick B. 2002. 99-115. Environmental Health Perspectives. Butera R. Reich A. Baden D. Lancaster J. Abraham W.. 825-34. 479-516. Locatelli C...R. 519-50.. 53. Botana L. Detection of brevetoxins in the twenty-first century. Botana L.S. Schrank K. Abbott J.FISH AND SHELLFISH TOXINS 65. Whitney P. Freidman M. 77...J. Naar J. Competitive ELISA to detect brevetoxins from Karenia brevis (formerly Gymnodinium breve) in seawater. Neurologic Shellfish Poisoning (NSP). Wolny J. in Seafood and Freshwater Toxins.. Reich A... 465-70. in FAO Food and Nutrition Paper 80.. Marine Biotoxins.M. Food and Agriculture Organisation.S.E.. 171 .H.M. 2007. Naar J. 50. Bienfang P. Kirkpatrick B. Fleming L. Cheng Y. 73.S. 117-71. 2007. Poon-King C. Zaias J.. 2007. Granade R. 707-23.
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.
quality assurance or control. 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. Conduct a hazard analysis. ingredient and packaging buyers. food microbiology.The application of methods.A point. 5. 8. hygiene managers. 8. Verification .1 is a flow diagram illustrating this 12-stage logic sequence. Each stage is discussed below in detail. from primary production to final consumption. Figure 8. operation or stage in the food chain including raw materials. Establish documentation concerning all procedures and records appropriate to these principles and their application. The team should also include 177 . Determine the CCPs. in addition to monitoring to determine compliance with the HACCP plan.HACCP IN FISH AND SEAFOOD PRODUCT MANUFACTURE Step . Establish the corrective action to be taken when monitoring shows that a CCP is not under control. 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.3 Stages of a HACCP Study The HACCP system consists of the following seven basic principles: 1. 4.1 Assemble HACCP team HACCP requires management commitment of resources to the process. 7. 6. procedures.Obtaining evidence that the elements of the HACCP plan are effective. Establish a system to monitor control of the CCP. tests and other evaluations. food chemistry or engineering. procedure. An effective HACCP plan is best carried out as a multidisciplinary team exercise to ensure that the appropriate product-specific expertise is available. 2. Validation . Establish procedures for verification to confirm that the HACCP system is working effectively. Establish the critical limit(s). 3.3.
chemical and/or physical).FISH AND SEAFOOD personnel who are involved with the variability and limitations of the operations. biological. it may be obtained from external sources. Fig. 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.g.1. 8. Logic sequence for application of HACCP 178 .
1 Fish and seafood Certain products may be contaminated or carry pathogenic organisms as a part of the natural flora.4 Construct a flow diagram The flow diagram should be constructed by the HACCP team and should contain sufficient technical data for the study to progress. It should provide an accurate representation of all steps in the production process from raw materials to the end179 . heat treatment. fermentation. storage and distribution conditions and instructions for use. or in aquaculture operations.1.3. such as enteric viruses in shellfish.1 Fish and seafood The HACCP team should ideally have access to expertise on the appropriate fisheries. contaminated fish and shellfish should not be used. pH. packaging.g. If the processing does not include a kill step. However. naturally present on fish from warm waters (> 15 °C). etc. In addition.3.g.). 8. 8. shelf life. including practices onboard fishing vessels. it must be noted here that certain toxins (algal and shellfish) are not destroyed by heat treatments. challenge testing and inoculation studies for evaluation of safety aspects are therefore essential for an effective HACCP team. which should be described in detail. aquaculture drug residues and potentially pathogenic Vibrio spp. marine toxins associated with particular fish and shellfish species. 8. freezing.3.3.3 Identify intended use The intended use should be based on the expected uses of the product by the enduser or consumer (e. such as children or the elderly. physical and chemical structure (including water activity (aw).). These factors are likely to have a significant effect on some potential hazards. is a cooking process required?). Vulnerable groups of the population. the only CCP that can render the product safe from pathogenic organisms is adequate heat treatment during preparation.3. composition. The description should include information such as the product name.HACCP IN FISH AND SEAFOOD PRODUCT MANUFACTURE 8. processing conditions (e.3. 8. for example contamination by foodborne pathogens as a result of time/temperature abuse or unsatisfactory handling practice. there may be species-specific hazards related to certain fish and shellfish. Expertise on such inherent hazards.2 Describe the product It is important to have a complete understanding of the product. etc. may need to be considered specifically. It is also important to identify the consumer target groups.
However. and any discrepancies between the flow diagram and normal practice must be recorded and the diagram amended accordingly. 8. and therefore a detailed description of the procedure applied must be available. If HACCP studies are applied to proposed new process lines or products. Ideally.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. conduct a hazard analysis. features of equipment design. 8. For example.FISH AND SEAFOOD product. It may include details of the factory and equipment layout. The analysis is likely to include consideration of: ● the likely occurrence of hazards and the severity of their adverse health effects. The operation should be observed at all stages.1 Fish and seafood Examples of flow diagrams for specific fish products may be found in the appropriate product chapters. the qualitative and/or quantitative evaluation of the presence of hazards. It is also important to include observation of production outside normal working hours such as shift patterns and weekend working. it is essential that the details of each step are fully appreciated and recorded. Many fish-processing operations have relatively few steps and the flow diagrams appear simple. 8.6 List all potential hazards associated with each step. ingredient specifications.4.3. it should also include details of CCP steps. or circumstances of any reclaim or rework activity. once determined. cleaning and hygiene procedures and storage conditions. It is essential that the diagram is accurate. 180 ● ● . survival or multiplication of pathogenic microorganisms. The team should then conduct a hazard analysis to identify which hazards are of such a nature that their elimination or reduction to an acceptable level is essential to the production of safe food. adequate chilling of finfish immediately after capture is vital for the control of several potential hazards. 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. then any pre-drawn HACCP plans must be reviewed once the lines/products are finalised. time/temperature data.3.3. because the hazard analysis and decisions regarding CCPs are based on these data.
g. The HACCP team should then determine what control measures exist that can be applied for each hazard. 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. Hazards specific to certain types of product are detailed in the appropriate chapters of this manual. or may originate from polluted water or from post-capture contamination (e. The incidence of psychrotrophic. non-proteolytic Clostridium botulinum on fresh fish is also sufficiently high that its presence may be assumed. 181 . there are particular hazards associated with contamination of shellfish by microorganisms from human sewage. to generalise about expected hazards and the reader is once again referred to the appropriate product chapter for additional advice on specific hazards. any food safety objectives or manufacturer’s food safety requirements. and the potential growth of Listeria monocytogenes on smoked fish.3. since this may interfere with the analysis. Salmonella spp.g. Pathogens may be part of the resident microflora of the living animal (e. One control measure may be relevant to several process steps. the number of consumers potentially exposed and their vulnerability.1 Fish and seafood The term 'fish and seafood' includes an extremely varied group of products.). or desirable. modified-atmosphere packaging (MAP) of chilled raw fish may provide conditions suitable for the growth of psychrotrophic C. or poor control of batter mixes in frozen battered fish portions may allow growth of Staphylococcus aureus and production of enterotoxin. 8. 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. Vibrio spp. Many of the microbiological hazards associated with fish and seafood products are derived from the raw materials. Note: it is important at this stage that no attempt is made to identify CCPs. botulinum. and there are an equally varied range of potential hazards associated with them. For example. and viruses). Therefore it is not possible. For example.6. the hurdle effect. Hazards introduced during processing of fish products depend very much on the characteristics of the process. where a hazard is repeated.HACCP IN FISH AND SEAFOOD PRODUCT MANUFACTURE ● ● ● ● production or persistence of toxins.
and the reader is referred to the appropriate product chapter. biological or sensory parameters. 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. 8. and its use may not always be appropriate. structured approach to decision making. it is necessary to specify both time and temperature for a thermal process. application of the decision tree should be flexible.7 Determine CCPs The determination of CCPs in the HACCP system is facilitated by a decision tree (Figure 8.7.8 Careful selection of sources for raw materials Adequate temperature control Effective sanitation Food handler hygiene Prevention of cross-contamination Establish critical limits for each CCP Critical limits separate acceptable from unacceptable products. 8. 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 . critical limits should be specified and validated for each CCP.3. Criteria used to set critical limits must be measurable and may include physical.3. then the process must be modified to include a control measure. However. given the enormous variety of fish products and processes in use. but for which no control exists at that step or any other. If a significant hazard has been identified at a step where control is required for safety. it can be said that effective control measures are likely to include the following: • • • • • 8. Where possible.FISH AND SEAFOOD 8.8.1 Fish and seafood Again.2) to provide a logical. However. It is also essential that the HACCP team has access to sufficient technical data to determine the CCPs effectively. More than one critical limit may be defined for a single step.3.3. chemical. it is not possible to generalise on likely CCPs.1 Fish and seafood Specific product chapters provide information on criteria that may be used to set critical limits. For example.
HACCP IN FISH AND SEAFOOD PRODUCT MANUFACTURE
• • • 8.3.9
Water activity (aw) values for dried fish pH values in fermented fish Microbiological quality of water in shellfish production areas Establish a monitoring system for each CCP
Monitoring involves planned measurement or observation of a CCP relative to its critical limits. Monitoring procedures for target levels must be able to detect loss of control of the CCP, and should provide this information with sufficient speed to allow adjustments to be made to the control of the process before the critical limits are violated. Monitoring at critical limits should be able to rapidly detect when the critical limit has been exceeded. Monitoring should therefore either be continuous, or carried out sufficiently frequently to ensure control at the CCP. Therefore, physical and chemical on-line measurements are usually preferred to lengthy microbiological testing. However, certain rapid methods, such as ATP assay by bioluminescence, may be useful for assessment of adequate cleaning, which could be a critical limit for some CCPs, for example pre-start-up hygiene. Persons engaged in monitoring activities must have sufficient knowledge, training and authority to act effectively on the basis of the data collected. These data should also be properly recorded. 8.3.10 Establish corrective actions
For each CCP in the HACCP plan, there must be specified corrective actions to be applied if the CCP is not under control. If monitoring indicates a deviation from the critical limit for a CCP, action must be taken that will bring it back under control. Actions taken should also include proper isolation of the affected product, repair of defective equipment, and an investigation into why the deviation occurred. A further set of corrective actions should relate to the target levels, if process drift is occurring. In this case, only repair of the process defect and investigation of the fault are required. All corrective actions should be properly recorded. 8.3.11 Establish verification procedures
Verification usually involves auditing and testing procedures. Auditing methods, procedures and tests should be used frequently enough to determine whether the HACCP system is being followed, and is effective at controlling the hazards. These may include random sampling and analysis, including microbiological testing. Although microbiological analysis is generally too slow for monitoring purposes, it can be of great value in verification, since many of the identified hazards are likely to be microbiological. In addition, reviews of HACCP records are important for verification purposes. These should confirm that CCPs are under control and should indicate the nature
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of any deviations and the actions that were taken in each case. It is also useful to review customer returns and complaints regularly. 8.3.12 Establish documentation and record keeping
Efficient and accurate record keeping is an essential element of a HACCP system. The procedures in the HACCP system should be documented. ● ● Determination of critical limits The completed HACCP plan
Examples of documented procedures include: ● ● The hazard analysis Determination of CCPs
Examples of recorded data include: ● ● ● Results of monitoring procedures Deviations from critical limits and corrective actions Records of certain verification activities, e.g. observations of monitoring activities, and calibration of equipment.
The degree of documentation required will depend partly on the size and complexity of the operation, but it is unlikely to be possible to demonstrate that an effective HACCP system is present without adequate documentation and records. The length of time that records are kept will be as per company policy, but should not be less than one year beyond the shelf life of the product. Three to five years is typical for many food companies. 8.4 Implementation and review of the HACCP plan
The completed plan can only be implemented successfully with the full support and co-operation of management and the workforce. Adequate training is essential and the responsibilities and tasks of the operating personnel at each CCP must be clearly defined.
HACCP IN FISH AND SEAFOOD PRODUCT MANUFACTURE
Fig. 8.2. CCP Decision Tree (Adapted from Codex Alimentarius Commission, 2003) Finally, it is essential that the HACCP plan be reviewed following any changes to the process, including changes to raw materials, processing conditions or equipment, packaging, cleaning procedures and any other factor that may have an effect on product safety. Even a small alteration to the product or process may invalidate the HACCP plan and introduce potential hazards. Therefore, the implications of any changes to the overall HACCP system must be fully
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considered and documented, and adjustments made to the procedures as necessary. Triggered reviews/audits should occur as a result of changes, whereas a scheduled review/audit should be annual, as a minimum. 8.5 References
Wareing P.W., Carnell A.C. HACCP – A Toolkit for Implementation. Leatherhead, Leatherhead Food International. 2007. Bernard D., Scott V. Hazard Analysis and Critical Control Point System: use in controlling microbiological hazards, in Food Microbiology: Fundamentals and Frontiers. Eds. Doyle M.P., Beuchat L.R. Washington D.C., ASM Press. 2007, 971-86. Su Y-C., Daeschel M.A. Sanitation and HACCP, in Surimi and Surimi Seafood. Ed. Park J.W. Boca Raton, CRC Press. 2005, 141-62. Huss H.H. Considerations in the application of the HACCP principles to seafood production, in Assessment and Management of Seafood Safety and Quality. Eds. Food and Agricultural Organisation, Huss H.H., Ababouch L., Gram L. Rome, FAO. 2004, 153-77. Chen H-C., Chang P.C-M. Frozen seafood safety and HACCP, in Handbook of Frozen Foods. Ed. Hui Y.H., Cornillon P., Legaretta I.G., Lim M.H., Murrell K.D., Nip WK. New York, Marcel Dekker. 2004, 339-63. Mortimore S., Mayes T. The effective implementation of HACCP systems in food processing, in Foodborne Pathogens: Hazards, Risk Analysis and Control. Eds. Blackburn C.deW., McClure P.J. Cambridge, Woodhead Publishing Ltd. 2002, 229-56. Mayes T., Mortimore C.A. Making the Most of HACCP: Learning from Other’s Experience. Cambridge, Woodhead Publishing. 2001. Mortimore S.E., Wallace C., Cassianos C. HACCP (Executive Briefing). London, Blackwell Science Ltd. 2001. Chartered Institute of Environmental Health. HACCP in Practice. London, Chadwick House Group Ltd. 2000. Jouve J.L. Good manufacturing practice, HACCP, and quality systems, in The Microbiological Safety and Quality of Food, Volume 2. Eds. Lund B.M., BairdParker T.C., Gould G.W. Gaithersburg, Aspen Publishers. 2000, 1627-55. Stevenson K.E., Bernard D.T. HACCP: A Systematic Approach to Food Safety. Washington D.C., Food Processors Institute. 1999. National Seafood HACCP Alliance. IQF cooked shrimp processing HACCP model, in HACCP User's Manual. Ed. Corlett D.A. Gaithersburg, Aspen Publishers. 1998, 454-73. Mortimore S., Wallace C. HACCP: A Practical Approach. Gaithersburg, Aspen Publishers. 1998.
HACCP IN FISH AND SEAFOOD PRODUCT MANUFACTURE Khandke S.S., Mayes T. HACCP implementation: a practical guide to the implementation of the HACCP plan. Food Control, 1998, 9 (2-3), 103-9. Forsythe S.J., Hayes P.R. Food Hygiene, Microbiology and HACCP. Gaithersburg. Aspen Publishers. 1998. Food and Agriculture Organisation. Food Quality and Safety Systems: A Training Manual on Food Hygiene and the Hazard Analysis and Critical Control Point (HACCP) System. Rome, FAO. 1998. US Food and Drug Administration. Fish and Fishery Products Hazards and Controls Guide. Rockville, Food Drug Administration. 1998. National Advisory Committee on Microbiological Criteria for Foods. Hazard Analysis and Critical Control Point Principles and Application Guidelines. 1997. Gardner L.A. Testing to fulfil HACCP (Hazard Analysis Critical Control Points) requirements: principles and examples. Journal of Dairy Science, 1997, 80 (12), 3453-7. Martin R.E., Collette R.L, Slavin J.W. Fish Inspection, Quality Control and HACCP: A Global Focus. Proceedings of a Conference, Arlington, May 1996. Lancaster P.A., Technomic Publishers. 1997. Codex Alimentarius Commission. Hazard Analysis Critical Control Point (HACCP) System and guidelines for its application, in Food Hygiene: Basic Texts. Ed. Codex Alimentarius Commission. Rome, FAO. 2003, 21-31. Garrett E.S., Jahncke M.l., Tennyson J.M. Microbiological hazards and emerging food safety issues associated with seafoods. Journal of Food Protection, 1997, 60 (11), 1409-15. National Seafood HACCP Alliance. HACCP: Hazard Analysis and Critical Control Point Training Curriculum. Raleigh, North Carolina State University. 1997. Dillon M., McEachern V. HACCP and quality assurance of seafood, in Fish Processing Technology. Ed. Hall G.M. London, Blackie. 1997, 224-48. Savage R.A. Hazard Analysis Critical Control Point: a review. Food Reviews International, 1995, 11 (4), 575-95. Pearson A.M., Dutson T.R. HACCP in Meat, Poultry and Fish Processing. Glasgow, Blackie. 1995. Huss H.H., Food and Agriculture Organisation. Assurance of Seafood Quality. FAO Fisheries Technical Paper No. 334. Rome, FAO. 1994. Bonnell A.D. Process control, in Quality Assurance in Seafood Processing: A Practical Guide. Ed. Bonnell A.D. London, Chapman & Hall. 1994, 39-53. Pierson M.D., Corlett D.A., Institute of Food Technologists. HACCP: Principles and Applications. New York, Van Nostrand Reinhold. 1992. Hudak-Roos M., Garrett E.S. Monitoring, corrective actions and record keeping in HACCP, in Quality Assurance in the Fish Industry: Proceedings of an International Conference, Copenhagen, August 1991. Eds. Huss H.H., Jakobsen M., Liston J. Amsterdam, Elsevier. 1992, 521-31.
188 . Volume 4: Application of the Hazard Analysis Critical Control Point (HACCP) System to Ensure Microbiological Safety and Quality.L.H. 1992. Geneva. Food Control. 1992. International Commission on Microbiological Specifications for Foods. 3 (1). Bryan F.. World Health Organisation.HACCP IN FISH AND SEAFOOD PRODUCT MANUFACTURE Huss H. 1992. 15 (1/2). 14-9. Development and use of the HACCP concept in fish processing. International Journal of Food Microbiology. Blackwell Scientific Publications. Microorganisms in Foods. 1988. Simple users' guide to the hazard analysis critical control point concept for the control of food microbiological safety. Hazard Analysis Critical Control Point Evaluations: A Guide to Identifying Hazards and Assessing Risks Associated with Food Preparation and Storage. 33-44. Oxford. Mayes T. WHO.
final product specifications. have been replaced by several linked measures on food safety rules and associated animal health controls. from structural conditions in the factory or process facility. Microbiological standards have a useful role and help establish requirements for the microbiological safety and quality of food and raw materials.1 Introduction Hygiene is an important aspect of ensuring food safety. Jenny Pfleger Leatherhead Food International Randalls Road Leatherhead Surrey KT22 7RY United Kingdom 9. with the intention of introducing consistency and clarity throughout the food production chain from primary production. including microbiological criteria. to sale or supply to the final consumer. however. Procedures under the new legislation are based on risk assessment and management and follow a 'farm to fork' approach to food 189 . 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.9. to personnel requirements. A new package of European Commission (EC) hygiene measures became applicable on 1 January 2006 to update and consolidate the earlier 17 hygiene directives. A number of standards are provided in food legislation. the existence of microbiological standards alone cannot protect consumer health. and conditions of raw materials. The general food hygiene Directive 93/43/EEC and other Directives on the hygiene of foodstuffs and the health conditions for the production and placing on the market of certain products of animal origin intended for human consumption. under which food is produced to optimise public health and to prevent. eliminate or acceptably control pathogen contamination of food. EC FOOD HYGIENE LEGISLATION Eugenia Choi and Dr. The new legislation was designed to establish conditions. Hygiene is a general concept that covers a wide subject area. and one that plays an important role in most countries’ food legislation. transport and delivery vehicles requirements.
p. Regulation 854/2004 relates to the organisation of official controls on products of animal origin. the following European Union (EU) hygiene regulations have applied: • • • Regulation (EC) No. 2073/2005 on microbiological criteria for foodstuffs • The general hygiene requirements for all food business operators are laid down in Regulation 852/2004. J. European Communities L 268.B. p. N.1). storing. 190 • • .FISH AND SEAFOOD safety with the inclusion of primary production in food hygiene legislation.7. transportation.1992. 853/2004 of the European Parliament and of the Council laying down specific hygiene rules for food of animal origin 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) No. and sets out what those enforcing the provisions have to do. 9. as last amended by Regulation (EC) No 806/2003. Regulation 853/2004 supplements Regulation 852/2004 in that it lays down specific requirements for food businesses dealing with foods of animal origin. 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. 852/2004 of the European Parliament and of the Council on the hygiene of foodstuffs Regulation (EC) No. packaging. 7.1991. distribution.2 Legislative Structure From 1 January 2006.9. 41). 24. 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. handling and offering for sale or supply to the consumer. p.1991. 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. 24. manufacturing. processing. Subsequently.15). A number of more detailed implementing and transitional measures have been adopted at EC level. Prescribed are detailed measures to ensure the safety and wholesomeness of food during preparation.9.
with primary production.LEGISLATION • Council Directive 93/43/EEC of 14 June 1993 on the hygiene of foodstuffs (OJ L 175. There are some exemptions. Wales and Northern Ireland. national legislation in the form of a Statutory Instrument (S.) in England. This Regulation lays down general rules for food business operators on the hygiene of foodstuffs. and binding in its entirety on all member states from the date of entry into force. Wales and Northern Ireland). setting offences. right through to the implementation of procedures based on HACCP principles. and also if the producer supplies small amounts of primary product to the final consumer or local retail establishments supplying the final consumer. and of hunting and fishing. processing and distribution of food under their control satisfy the relevant hygiene requirements laid down in Regulation (EC) No. is required to give effect to the EU regulations. 191 .11) The EU hygiene regulations apply to all stages of food production including primary production.I. for example. 19. as amended) came into force on 11 January 2006 (separate but similar national legislation also came into force on that day in Scotland. 2073/2005. 9. The Food Hygiene (England) Regulations 2006 (S. domestic preparation or handling. Additionally the requirements of EC Regulation 853/2004 must be complied with. 1 . the legislation is directly applicable law. As regulations. and equivalent legislation in Scotland. revocation of existing implementing legislation. 2006 No. 852/2004 on the General Hygiene of Foodstuffs Food business operators must ensure that all stages of production. penalties and powers of entry. The national legislation in all four UK countries also applied the provisions of the EU Microbiological Criteria Regulation No. 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.3 Regulation (EC) No.g. p. e.14. of stock farming.7. Although EU food hygiene regulations are directly applicable in the individual Member States there are some aspects where Member States are required or allowed to adopt certain provisions into their national laws. Although the regulations have the force of law.1993. food storage that is for private or domestic consumption. 852/2004.I. which will be covered later in this chapter. Primary products are defined as products of primary production including products of the soil. particularly taking into account a number of factors ranging from ensuring food safety throughout the food chain to begin with primary production. The regulation lays down general hygiene provisions for which food business operators carrying out primary production must comply with as laid down in Part A of Annex 1. etc.
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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: 126.96.36.199 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.
188.8.131.52 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.
184.108.40.206 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.
220.127.116.11 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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18.104.22.168 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.
22.214.171.124 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.
126.96.36.199 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.
188.8.131.52 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. 184.108.40.206 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.
220.127.116.11 Chapter X Chapter X lays down provisions applicable to the wrapping and packaging of foodstuffs to avoid contamination of any form. 18.104.22.168 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). 22.214.171.124 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:
Factory vessel means any vessel on board which fishery products undergo one or more of the following operations followed by wrapping or packaging and. gutting and removal of fins and. purification centres or dispatch centres in tanks or any other installation containing clean seawater. whether whole or prepared. followed by wrapping or packaging. skinning. filleting. after preparatory work such as bleeding. parts and products of such animals. heading. Conditioning means the storage of live bivalve molluscs coming from class A production areas. 197 . where appropriate. slicing. 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. Bivalve molluscs means filter-feeding lamellibranch molluscs Marine biotoxins means poisonous substances accumulated by bivalve molluscs. or in natural sites. Fresh fishery products means unprocessed fishery products. such as gutting. and chopping. slicing. 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. to preserve or to improve organoleptic qualities and to ensure they are in a good state of vitality before wrapping or packing. 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. where necessary. to remove mud. shelling. and all mammals.LEGISLATION Fishery products means all seawater or freshwater animals (except for live bivalve molluscs. including products packaged under vacuum or in a modifiedatmosphere that have not undergone any treatment to ensure preservation other than chilling. if necessary: chilling or freezing: filleting. in particular as a result of feeding on plankton containing toxins. mincing or processing. heading. slime or sand. shucking. live tunicates and live marine gastropods. Freezer vessel means any vessel on board in which freezing of fishery products is carried out.
9. the food business operator may apply the same identification mark to both types of food. for other codes. the code would be UK. 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. For the UK.4.1.1 Application of the identification mark (i) The identification mark must be applied before the product leaves the establishment of production. Section I of Regulation (EC) 853/2004 are given as follows: 9. the mark must be oval shaped and include the abbreviation CE. please refer to Regulation (EC) 853/2004.1. ES. EB or WE.4. EC. EÜ.4. EK. when a product's packaging and/or wrapping is removed or is further processed in another establishment. In this case. EG. (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. the mark must show the approval number of the establishment.1 Identification marking An identification mark needs to be applied following the requirements of Annex II.2 Form of the identification mark (i) The mark must be legible and indelible.4. (iv) If the identification mark is applied in an establishment within the Community. (iii) Additionally.2 Fishery product requirements 9.2. EF. and the characters easily decipherable and be clearly displayed. EO. EK.2.2. (ii) However. a new mark must be applied to the product. EY. Section I of Regulation (EC) 853/2004.FISH AND SEAFOOD 9. 198 . If an establishment manufactures both food to which Regulation 853/2004 applies and food to which it does not. Extracts of the requirements of Annex II. the new mark must show the approval number of the establishment where this occurred. (ii) The mark must indicate the name of the country in which the establishment is located and written out in full or shown as a two-letter code in accordance with the relevant ISO standard.
2 Hygiene requirements for fishery products The regulation details specific hygiene requirements for fishery products. reference should be made to the actual Regulation. Extracts of the requirements of Annex III. including vessels. processing. its wrapping or the packaging. a) requirements for establishments. (v) When the mark is applied directly to products of animal origin. involved in primary production and such operations accompany the needs of Annex I of Regulation (EC) 852/2004 b) for other establishments including vessels. C and D.1. it applies to those animals when not placed on the market live. Chapter IV. This section does not apply to bivalve molluscs. Therefore. and are intended for further handling.e. Section VIII of Regulation 853/2004. i.2. or be printed on a label affixed to the product. it is sufficient to apply the mark to the exterior of that package only. (iii) In the case of liquid. for full requirements. granulate and powdered products of animal origin carried in bulk. the colourings used must be EC authorised colours.2. With the exception of Chapters I and II.3 Method of marking (i) The mark is to be applied directly to the product. It may also be a tag made of a resistant material affixed in such a way that it cannot be removed. the mark may be applied to the outer surface of the container or packaging. wrapping or packaging in another establishment. (iv) If products of animal origin are placed in a package destined for direct supply to the final consumer. echinoderms. The requirements of this section accompany those in Regulation (EC) 852/2004. and fishery products carried in bulk. as amended.4. they would need to be obtained following Section VII on live bivalve molluscs. Parts A. tunicates and marine gastropods when placed on the market live. the wrapping or the packaging. the requirements accompany those of Annex II of Regulation (EC) 852/2004 199 . (ii) For products of animal origin that are placed in transport containers or large packages. specifically relating to fishery products are given below.4. an identification mark is not necessary if accompanying documentation contains the name of the country or its two-letter code. the approval number of the establishment and the abbreviation for the community where appropriate.LEGISLATION 9. 9. Part A and Chapter V apply to retail. Chapter III.
refrigeration and wrapping.) 9. clean seawater may be used for the handling and washing of fishery products. bleeding. this supplements the requirements of Annex II. removing fins. heading. A. they also include (i) the transport and storage of fishery products that have not been substantially altered (including live fishery products. if a vessel has water coming in that is used with fishery products. Requirements for vessels designed and equipped to preserve fresh fishery products for more than 24 hours 200 ..2. it must be placed somewhere that will not contaminate the water supply. gutting. 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. from the place of production to the first establishment of destination. within fish farms on land) (ii) the transport of fishery products that have not been substantially altered (including live fishery products. surfaces that contact fishery products need to be made from cleanable corrosion-resistant material. 2.4. and (b) related operations include any of the following operations if done on board fishing vessels: slaughter. i. Structural and equipment requirements refer to the following points 1.e. (a) primary production covers the farming. Chapter VII of Regulation 852/2004. Requirements for all vessels Vessels should be made in a way that does not cause contamination with substances such as fuel and oil etc. fishing and collection of live fishery products with a view to their being placed on the market. the production of ice used to chill fishery products and the rapid cooling of crustaceans and molluscs after their cooking.FISH AND SEAFOOD c) for the water supply.2. The definition of fishery products covers primary production as well as its associated operations. There is a derogation from point a.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.1 Chapter I . or to handle or process them after harvesting.
Storage areas for the finished products also need to be sufficiently large and cleanable. and not more than 0 °C after 16 hours. A temperature recording device must be present and placed where the temperature in the hold is the highest. Such devices must achieve a chilling rate that ensures that the mix of fish and clean seawater does not exceed 3 °C six hours after loading. tanks must incorporate devices that allow a uniform temperature throughout the tanks. 201 . where necessary. (iii) In vessels equipped for chilling fishery products in cooled clean seawater. (v) Special equipment for disposing of waste/ inedible fishery products into the sea or a watertight tank and allocation of separate areas. 3. (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. any waste processing unit needs to have a separate hold to store the waste. (iii) The above requirements for holds in vessels designed and equipped to preserve fresh fishery products for more than 24 hours must be met. (iv) There needs to be somewhere for storing packaging materials and this is to be separate from product preparation and processing areas. to ensure that melt water does not stay in contact with the products. (ii) Refrigeration equipment needs to keep fishery products in storage holds at not more than -18 °C. tanks or containers for the storage of fishery products at the temperatures laid down in Chapter VII. where necessary. Requirements for freezer vessels (i) These need to contain freezing equipment in order to achieve a core temperature of no more than -18 °C.LEGISLATION (i) Such vessels need to have holds. 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. 4. recording of temperatures. and allow the monitoring and. Holds and containers used for storing fishery products must allow its preservation under adequate hygiene conditions and.
Livers and roes for human consumption must be preserved under ice (temperature to approach that of melting ice) or be frozen. e) In cases where whole fish intended for canning is frozen in brine. do not have to meet the above requirements if no other form of handling or processing takes place on board such vessels. the product must be kept at no more than -9 °C. b) Factory vessels on board which crustaceans and molluscs are cooked. B.Requirements during and after landing 9.2. 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. d) Any gutting or heading of fish on board is to be done hygienically and quickly after capture. other than those kept alive. c) Fishery products. and products washed thoroughly. where fishery products are displayed for sale. 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. landed as soon as possible. Handlers may use spiked instruments to move large fish or fish that might injure them. must be chilled or. provided no damage occurs to the flesh. c) Factory vessels that freeze fishery products must have equipment meeting the requirements for freezer vessels as stated above. 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. Chapter II . chilled and wrapped.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. (ii) In the case of food business operators responsible for auction and wholesale markets etc.4. b) Fishery products are to be handled and stored in a way that prevents the flesh bruising.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. Parts that may present a danger to public health are to be removed quickly and kept away from products for human consumption.
3 Chapter III . prepared or processed immediately after arrival at an establishment on land. (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. and where necessary. they must be stored under ice in appropriate facilities. Additionally. Requirements for fresh fishery products (i) Where chilled or unpackaged products are not distributed.Requirements for establishments. Requirements for mechanically separated fishery products (i) Raw materials used must satisfy the following 203 . points 1 and 2. Food business operators need to co-operate with the relevant competent authorities so as to allow them to carry out official controls as laid down in Regulation (EC) No. 9. 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. 854/2004. (iv) Filleting and cutting is to be carried out in a way that avoids contamination or spoilage of fillets or slices. including vessels. They should be wrapped. part I. dispatched. handling fishery products A. C. packaged and chilled as quickly as possible after preparation. B. Gutting should ideally be done quickly once products have been caught or landed and afterwards be washed thoroughly.2. 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. C. Fillets and slices must then be removed from work tables as soon as possible after preparation.4. (iii) Whole and gutted fresh fishery products can be transported and stored in cooled water on board vessels. Chapter I.LEGISLATION consumption. Packaged fresh fishery products must be chilled to a temperature approaching that of melting ice. (ii) Heading and gutting must be carried out hygienically. there are requirements that such premises should be well lit and vehicles emitting fumes should not be able to access the premises.2.
stating the type of process they have undergone. (iv) (wild) Atlantic and Pacific salmon. Requirements concerning parasites (i) The fishery products below must be frozen at no more than . or a stabilising treatment. (iii) A document from the manufacturer. they must be cooled rapidly. 9. 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. (ii) The manufacturing process must satisfy the following Mechanical separation is to be carried out immediately after filleting. If no other method of preservation is used. Once produced. cooling must continue until a temperature 204 . mechanically separated fishery products must be frozen as quickly as possible or incorporated into a product intended for freezing.Requirements for certain processed fishery products A. D.FISH AND SEAFOOD Only whole fish and bones after filleting may be used to produce mechanically separated fishery products.4. except when supplied to the final consumer. must accompany fishery products referred to above when placed on the market.2.2. and whole fish must be gutted and washed beforehand. (ii) mackerel.4 Chapter IV . and (c) marinated and/or salted fishery products. (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. raw materials must have no guts. if the processing is insufficient to destroy nematode larvae.20 °C in all parts of the product for at least 24 hours (this treatment must be applied to the raw product or the finished product): (a) fishery products to be consumed raw or almost raw. Requirements for cooking of crustaceans and molluscs After crustaceans and molluscs are cooked. (iii) sprat. (b) fishery products from the following species.
B.4.5 Chapter V . raw materials need to be chilled as soon as possible and should meet the temperature requirements in Chapter VII. Requirements for fish oil intended for human consumption (i) In relation to the raw materials used in their preparation and their transport. 2074/2005).LEGISLATION approaching that of melting ice is reached. provided that the freshness criteria are met and the total volatile basic nitrogen (TVB-N) value of the unprocessed fishery products does not exceed the limits set out in point 1 of Chapter I of Section II of Annex II to Regulation (EC) No. C.2. the following conditions need to be met: A. 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). B. Shelling/shucking is to be done hygienically. 205 . 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. Histamine Histamine limits must not be exceeded (limits are set in Regulation (EC) No.2. 2073/2005). and the raw material is processed within 36 hours after loading. processing. Total volatile nitrogen Unprocessed fishery products cannot be placed on the market if chemical tests reveal that TVB-N or TMA-N limits are exceeded (limits are set in Regulation (EC) No.Health standards for fishery products In addition to compliance with microbiological criteria laid down in Regulation (EC) No. cooked products are to be frozen or chilled to the temperatures stated in Chapter VII. (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. However. refining and purification steps before being placed on the market for the final consumer. 2073/2005. centrifugation. 2074/2005. pressing. separation. 9. and after this.
7 Chapter VII . The requirements for histamine and parasites do not apply to whole fishery products being used directly for preparing fish oil intended for human consumption.2.4. thawed unprocessed fishery products. and when fishery products are wrapped on board fishing vessels. Molidae. 9. (ii) Frozen blocks prepared onboard vessels must be adequately wrapped before landing. Fresh. Toxins harmful to human health Fishery products derived from poisonous fish of the following families must not be placed on the market: Tetraodontidae. Fishery products contaminated with parasites may not be placed on the market for human consumption. In the case of whole frozen fish in brine intended for the manufacture of canned food. Diodontidae and Canthigasteridae.4. 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. 9.Storage of fishery products (i) Fresh fishery products.2.FISH AND SEAFOOD D. in particular Ruvettus pretiosus and Lepidocybium flavobrunneum.6 Chapter VI . 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. (ii) Frozen fishery products must be kept at a temperature of not more than -18 °C in all parts of the product. 206 .2. The scientific name must accompany the common name on the label.2. it should be kept at no more than -9 °C.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. prepared and processed fishery products belonging to the family Gempylidae. E. and cooked and chilled products from crustaceans and molluscs are to be kept at a temperature close to that of melting ice. Parasites Fishery products are to be visually examined so that visible parasites are identified before being placed on the market.
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.2. This covers Total volatile basic nitrogen (TVBN) limits and how to determine them.2. 9.4.). The temperature of frozen fishery products must not fluctuate above -15°C. species belonging to the Merlucciidae family. (b) 30 mg of nitrogen/100 g of flesh for the species belonging to the Pleuronectidae family (with the exception of halibut: Hippoglossus spp.LEGISLATION (iii) Fishery products kept alive are to be kept at a temperature which will not adversely affect food safety or its viability.2. (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 . 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. Helicolenus dactylopterus. 2074/2005. The required temperature for whole frozen fish in brine intended for the manufacture of canned food during transport is not stated.4. 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. provided that the journey is short and the competent authority allows this.Transport of fishery products The above points for Chapter VII apply in relation to transport. Sebastichthys capensis.8 Chapter VIII . melt water must not remain in contact with the products. (c) 35 mg of nitrogen/100 g of flesh for the species: Salmo salar. species belonging to the Gadidae family. 9.
1 Chapter I .4.FISH AND SEAFOOD Regulation (EC) No 853/2004. they supplement the requirements of Annex II to that Regulation. taken from at least three different points and mixed together by grinding. Apart from the provisions on purification. however. and point 3 of Chapter VII applies to retail. VI.3 The regulation details specific hygiene requirements for bivalve molluscs. The sample must consist of about 100 g of flesh. please refer directly to the regulation. for full requirements. Extracts of the requirements of Annex III. this Section also applies to live echinoderms. tunicates and marine gastropods. they supplement the requirements of Annex I to that Regulation. specifically relating to bivalve molluscs are given below. (b) In the case of other operations. Live bivalve mollusc requirements - 9. Section VII of Regulation 853/2004. The information required in the registration document is given in point 4 and includes the health status of the production area.4.3. 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. 9. Chapter IX applies to pectinidae harvested outside those areas. Member States may set limits at a higher level for certain species pending the establishment of specific Community legislation. where the raw material complies with points (a). 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. shellfish species etc. Chapters V. reference should be made to the actual regulation. as amended. (b) and (c) of Part B(1) of that Chapter. VIII and IX. A food business operator can only accept batch of such bivalve molluscs provided correct documentary requirements are presented. The copy of 208 . The requirements of this Section accompany those laid down in Regulation (EC) No 852/2004: (a) In the case of operations that take place before live bivalve molluscs arrive at a dispatch or purification centre.General requirements for the placing on the market of live bivalve molluscs Live bivalve molluscs can only be put on the market for retail sale if done through a dispatch centre.
Class C means relaying over a long time before it can be placed on the market For bivalve molluscs not subject to Class B or C treatments. Transport needs to allow sufficient drainage for best survival conditions.LEGISLATION document must be kept for at least 12 months post dispatch or receipt or longer as stated by the competent authority.2 Chapter II . Requirements for relaying live bivalve molluscs Only areas approved by the competent authority can be used for relaying live bivalve molluscs with a requirement that buoys. C. 9. Requirements for production areas (i) The competent authority classes (classed A. 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. B.Hygiene requirements for the production and harvesting of live bivalve molluscs A. 209 . B or C) production areas with fixed locations and boundaries for where live bivalve molluscs may be harvested and this must be complied with.4. (ii) Provided the requirements of chapter V are met. processing or relaying.Class A means they can be placed directly on the market .3. collection from: . poles or other fixed means are to identify the boundaries of the sites. Other requirements such as conditions for relaying in order to allow optimal conditions for purification and for permanent record keeping are also specified. Requirements for 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.Class B means treatment in a purification centre or after relaying is done before it can be placed on the market .
(iv) yessotoxins. B. 20 milligrams of domoic acid per kilogram (iii) okadaic acid. containers and a label to identify purification. 9. requirements if there are many batches of live bivalve molluscs. 210 .Health standards for live bivalve molluscs In addition to microbiological criteria and organoleptic characteristic requirements. dinophysistoxins and pectenotoxins together.4. Requirements for dispatch centres Handling should not result in contamination to any degree. shells of live bivalve molluscs are to be washed throughout with clean water and the molluscs are to come from one of the following: (i) Class A production area (ii) Relaying area (iii) Purification centre (iv) Another dispatch centre 9.4. 800 micrograms per kilogram.5 Chapter V .Structural requirements for dispatch and purification centres Premises on land should not be subject to flooding by tides etc. Requirements for purification centres There are requirements for live bivalve molluscs to be washed.3 Chapter III .FISH AND SEAFOOD 9. the levels of marine biotoxins should not exceed the following limits.3.3.Hygiene requirements for purification and dispatch centres A. for: (i) paralytic shellfish poison (PSP).3. 160 micrograms of okadaic acid equivalents per kilogram. and there are requirements for tanks and water storage containers.4.4 Chapter IV . 1 milligram of yessotoxin equivalent per kilogram. Before dispatch. the purification system to allow them to resume /maintain filter-feeding activity. (ii) amnesic shellfish poison (ASP).
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. the label. they cannot be re-immersed in or sprayed with water.9 Chapter IX . by way of derogation from directive 2000/13/EC.6 Chapter VI .Identification marking and labelling In addition to general identification marking (contained in 9.2.8 Chapter VIII . (Note: testing methods for detecting marine biotoxins are detailed in Annex III of Regulation (EC) 2074/2005) 9. (ii) Once the live bivalve molluscs have been packed for retail sale and have left the dispatch centre. 9.4. 160 micrograms of azaspiracid equivalents per kilogram. Requirements of Chapters III and IV are to be complied with. 9. dispatch centre or processing establishment. 211 .4.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. the species of bivalve mollusc and date of packaging (at least the date and month) are required.4. including the identification mark.3.Wrapping and packaging of live bivalve molluscs (i) Oysters must be wrapped or packaged with the concave shell downwards. (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. the date of minimum durability may be replaced by the words ‘these animals must be alive when sold’.3.3. Finally.7 Chapter VII .4.3. must be waterproof. The requirements for production areas as stated earlier apply to pectinidae and these can only be placed on the market through a fish auction.4.1). Also.LEGISLATION (v) azaspiracids. Registration document requirements also apply. 9.
The requirements refer to hygienic conditions and regular inspections both on land and at port to ensure compliance of temperature and other requirements. or (iii) bivalve molluscs. In this regulation. Fishery products in particular must comply with the requirements of Annex III of the regulation. (ii) fishery products not complying with the requirement concerning biotoxins. general principles for official controls in respect of all products of animal origin falling within the scope of the regulation are given.1 Regulation (EC) 854/2004 Annex III Chapter III states that fishery products are to be declared unfit for human consumption if: organoleptic.FISH AND SEAFOOD 9. chemical. The official controls include audits of good hygiene practices and hazard analysis and critical control point (HACCP)-based procedures. echinoderms. Chapter V of Regulation 853/2004 apply. Section VIII. tunicates or marine gastropods containing marine biotoxins in total quantities exceeding the limits referred to in Regulation (EC) No 853/2004. 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. physical or microbiological checks or checks for parasites have shown that they are not in compliance with the relevant Community legislation. 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. This refers to official controls or production and placing on the market of fishery products. 9.5 Regulation (EC) No. they derive from: (i) poisonous fish.5. It is a requirement that food business operators give assistance to ensure that official controls carried out by the competent authority can be done properly. or - - 212 . Therefore the health standards for fishery products as previously stated in Annex III.
Annex II 9. three dilutions Most Probable Number (MPN) test specified in ISO 16649-3. Requirements for the competent authority are stated in relation to the classification or a production or relaying area is stated. The reference method for this analysis is the five-tube. coli per 100 g of flesh and intravalvular liquid.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.600 E. 4.5. coli per 100 g of flesh and intravalvular liquid. live bivalve molluscs must not exceed 46. Also. The reference method for this analysis is the five-tube. - - - - B. Sampling plans and frequency are required. chapter II on hygiene requirements for the production and harvesting of live bivalve molluscs in Regulation (EC) 853/2004. In the remaining 10 % of samples. section VII.Official controls concerning live bivalve molluscs from classified production areas A. A summary is given as follows: 9. 000 E. Live bivalve molluscs from class A areas may be collected for direct human consumption provided health standards are met. as previously stated.2 This states official controls for the production and placing on the market of live bivalve molluscs. Monitoring of classified relaying and production areas This is to ensure there is no malpractice. Classification of production and relaying areas: This section reinforces that specified in Annex III. Regulation (EC) 854/2004. coli per 100 g of flesh and intravalvular liquid. Live bivalve molluscs from Class B areas must not exceed. that microbiological criteria and chemical contaminants amongst other requirements are met. three dilutions MPN test specified in ISO 16649-3.000 E. Live bivalve molluscs from Class C areas must not exceed 46. live echinoderms. chapter V is reinforced. live tunicates and live marine gastropods.2. in 90% of the samples. 213 .1 Chapter II .5.
5.2. which has applied since 1 January 2006. opening or closing a production area. 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.Official controls concerning pectinidae harvested outside classified production areas These are to be carried out in fish auctions.3 Imports By way of derogation from Article 11(1) of Regulation (EC) No 854/2004. Procedures for re-opening a closed production area are stated D.FISH AND SEAFOOD C. 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. F. 2073/2005. boundaries or class of a production area or its closure. 9. 9. establishes microbiological criteria for a range of foods. 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.5. 214 .6 Regulation (EC) No. 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. 2073/2005 on Microbiological Criteria for Foodstuffs Regulation (EC) No. E. the competent authority is to close the production area. Decisions after monitoring There are specific requirements whereby if sampling results indicate that health standards for molluscs are exceeded or there is a risk to human health.2 Chapter III . 9.
LEGISLATION The aim of this legislation is to complement food hygiene requirements. per unit(s) of mass. a batch of foodstuffs or a process. 215 . 9.4 referring to fishery products. applying to products placed on the market and process hygiene criteria that are applied during the manufacturing process. based on the absence.6. and/or on the quantity of their toxins or metabolites.2 Process hygiene criteria Chapter 2 focuses on process hygiene criteria. presence or number of microorganisms. with chapter 2. 9. The definition of ‘microbiological criterion’ means a criterion defining the acceptability of a product.II. The criteria are outlined in Table 9. and the legislation applies to all businesses involved in food production and handling.1 Food safety criteria Chapter 1 of the regulation focuses on food safety criteria that cover foods such as ready to eat foods and fishery products and live bivalve molluscs. volume. Two kinds of criteria have been established: food safety criteria.I. The relevant criteria are outlined in Table 9.6. area or batch. ensuring that foods being placed on the market do not pose a risk to human health.
monocytogenes.I Food safety criteria Sampling plan(1) n 10 5 Absence in 25 g EN/ISO 11290-1 c m M Products placed on the market during their shelf life Limit(2) Analytical reference method(3) Stage where the criterion applies Food category Microorganisms 1.16 Cooked crustaceans and molluscan shellfish Salmonella EN/ISO 6579 Products placed on the market during their shelf life .3 Ready-to-eat foods unable to support the growth of L.1 Ready-to eat foods intended for infants and ready-to-eat foods for special medical purposes (4) 5 0 100 cfu/g (5) EN/ISO 11290-2 (6) L.2 Ready-to-eat foods able to support the growth of L. monocytogenes L. monocytogenes 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. other than those intended for infants and for special medical purposes(4) (8) L. other than those intended for infants and for special medical purposes EN/ISO 11290-1 Products placed on the market during their shelf life Before the food has left the immediate control of the food business operator. monocytogenes. who has produced it EN/ISO 11290-2 (6) Products placed on the market during their shelf life 1.
17 Live bivalve molluscs and live echinoderms. tunicates and gastropods 1(16) 0 230 MPN/100 g of flesh and intravalvular liquid 100 mg/kg 200 mg/kg HPLC (19) ISO TS 16649-3 Salmonella 1.25 Live bivalve molluscs and live echinoderms.Food category Microorganisms Sampling plan(1) n 5 0 Absence in 25 g EN/ISO 6579 c m M Products placed on the market during their shelf life Products placed on the market during their shelf life Products placed on the market during their shelf life HPLC (19) Products placed on the market during their shelf life 400 g mg/kg Limit(2) Analytical reference method(3) Stage where the criterion applies 1.26 Fishery products from fish species associated with a high amount of histidine (17) 9 2 200 mg/kg Histamine LEGISLATION 217 1. manufactured from fish species associated with a high amount of histidine (17) Histamine . coli (15) 1. tunicates and gastropods 9 (18) 2 E.27 Fishery products which have undergone enzyme maturation treatment in brine.
subject to scientific justification. — fresh. Other categories of products can also belong to this category. including cocoa and chocolate products. Relevance of matrix effect in determination of biogenic amines in 6 7 8 15 16 17 18 19 218 . monocytogenes. that the product will not exceed the limit 100 cfu/g throughout the shelf life. In such a case the presumption laid down in Article 14(6) of Regulation (EC) No 178/2002. E. Engraulidae. Duflos G. 79. Products with pH ≤ 4. Scombresosidae.4 or aw ≤ 0.FISH AND SEAFOOD 1 2 3 4 n = number of units comprising the sample. honey and confectionery.92. Coryfenidae. soft drinks. beer. to the satisfaction of the competent authority. uncut and unprocessed vegetables and fruits. products with pH ≤ 5.25.. according to which the whole batch is to be deemed unsafe. c = number of sample units giving values between m and M For point 1. AOAC Internat. — bread. 2. Bouquelet S. excluding sprouted seeds. A pooled sample comprising a minimum of 10 individual animals. Pomatomidae. when he is not able to demonstrate. Malle P.0 and aw ≤ 0. Regular testing against the criterion is not required in normal circumstances for the following ready-to-eat foods: — those which have received heat treatment or other processing effective to eliminate L.1 . Particularly fish species of the families: Scombridae.. coli is used here as an indicator of faecal contamination. spirits and similar products.94. when recontamination is not possible after this treatment (for example. 5 This criterion shall apply if the manufacturer is able to demonstrate.1. The most recent edition of the standard shall be used.. that the product will not exceed the limit of 100 cfu/g throughout the shelf life. — — bottled or packed waters. This criterion shall apply to products before they have left the immediate control of the producing food business operator. Assay of biogenic amines involved in fish decomposition. biscuits and similar products. 1 ml of inoculum is plated on a Petri dish of 140 mm diameter or on three Petri dishes of 90 mm diameter. 1996. live bivalve molluscs. Malle P. References: 1. Dervin C. shall not apply. products with a shelf-life of less than five days shall be automatically considered to belong to this category. to the satisfaction of the competent authority. Valle M. Single samples may be taken at retail level. Bouquelet S. sugar. 43-9. cider. 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. J. wine. m = M.. products heat treated in their final package). Clupeidae..
monocytogenes in ready-to-eat foods intended for infants and for special medical purposes: — satisfactory. AOAC Internat. if the following requirements are fulfilled: 1. excluding live bivalve molluscs and live echinoderms. The test results demonstrate the microbiological quality of the batch tested(1). if all the values observed indicate the absence of the bacterium. if the presence of the bacterium is detected in any of the sample units. the mean value observed is ≤ m 2. Interpretation of the test results relating to Table 9.LEGISLATION plaice (Pleuronectes platessa) and whiting (Merlangus merlangus. Salmonella in different food categories: — satisfactory. 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. — unsatisfactory. if the presence of the bacterium is detected in any of the sample units. where the limit refers to a pooled sample. 1097-101. 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. — unsatisfactory. if all the values observed are ≤ the limit. no values observed exceed the limit of M.I The limits given refer to each sample unit tested. coli. if any of the values are > the limit. L. — unsatisfactory. J. monocytogenes in ready-to-eat foods able to support the growth of L. tunicates and gastropods in relation to testing E. if the presence of the bacterium is detected in any of the sample units. 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. (1) 219 . monocytogenes in other ready-to-eat foods: — satisfactory. Histamine in fishery products from fish species associated with a high amount of histidine: — satisfactory. — unsatisfactory. 1999. a maximum of c/n values observed are between m and M 3. if all the values observed indicate the absence of the bacterium. L. if all the values observed indicate the absence of the bacterium. 82. — unsatisfactory. L.
II Process hygiene criteria Limit(2) Analytical reference method(3) M 10 cfu/g ISO TS 16649-3 End of the manufacturing process End of the manufacturing process Improvements in production hygiene Stage where the criterion applies Action in case of unsatisfactory results Food category Microorganisms c 2 1 cfu/g m Sampling plan(1) n 2.4.TABLE 9. coli 5 shucked products of cooked crustaceans and molluscan shellfish 2 100 cfu/g 1000 cfu/g EN/ISO 6888-1 or 2 FISH AND SEAFOOD Coagulasepositive staphylococci 5 Improvements in production hygiene 220 .1 Shelled and E.
2006 No. The regulations provide for defences in relation to upward variations of the 8 °C temperature. The test results demonstrate the microbiological quality of the process tested. S. S. 2006 No. For example.7 Food Hygiene (England) Regulations 2006. — acceptable. The defendant may be required to produce well-founded scientific proof to support his claims. if all the values observed are ≤ m. if one or more of the values observed are > M or more than c/n values are between m and M. and the rest of the values observed are ≤ m. had not been 221 . the defendant will need to prove that the food was on service or display.LEGISLATION 1 n = number of units comprising the sample. Hot-held foods (food having been cooked or reheated and is for service or on display for sale) must not be kept below 63 °C. if a maximum of c/n values are between m and M. with chill holding tolerance periods. but there is also a general requirement that foods must not be kept at temperatures that would result in a risk to health. tolerance periods for chill-holding of foods and hot-holding variations. Schedule 4 of the Food Hygiene (England) Regulations 2006. Coagulase-positive staphylococci in shelled and cooked crustaceans and molluscan shellfish: — satisfactory. 14 details temperature control requirements for foods in general. coli in shelled and shucked products of cooked crustaceans and molluscan shellfish: — satisfactory.I. 14 (Hygiene requirements specific to the U.1 In the U. if all the values observed are ≤ m.II The limits given refer to each sample unit tested. — unsatisfactory.K. E. — acceptable. and the rest of the values observed are ≤ m. if a maximum of c/n values are between m and M.I. 9. which overrides the 8 °C requirement. and particularly that perishable foodstuffs must not be kept at above the maximum recommended storage temperature. 2 Interpretation of the test results relating to Table 9. if one or more of the values observed are > M or more than c/n values are between m and M. c = number of sample units giving values between m and M The most recent edition of the standard shall be used.K) Temperature control requirements 9. — unsatisfactory. The regulations prescribe a chilled food holding temperature of 8 °C or less.7.
S. The permitted reasons are given below: to facilitate handling during and after processing or preparation the defrosting of equipment.I. S. was kept at above 8 °C or the recommended temperature for an unavoidable reason.K. and was kept at above 8 °C or the recommended temperature. 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.I. 2006 No.8 Guidance In the U.gov. for a limited period consistent with food safety. it would need to be proved that the food was being transferred to or from a vehicle used for the activities of a food business. Recommended temperature: a specified temperature that has been recommended in accordance with a food business responsible for manufacturing. or. to or from premises (including vehicles) at which the food was to be kept at or below 8 °C or the recommended temperature. the period for which it can be expected to remain fit for sale when kept in a manner consistent with food safety. such as that below.FISH AND SEAFOOD previously put on display at more than 8 °C and had been kept there for less than four hours. 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. 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. For other food. 3). 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. Schedule 4 of the Food Hygiene (England) Regulations 2006 contains several definitions. Alternatively.food.uk/foodindustry/guidancenotes/hygguid/fhlguidance/ 222 . preparing or processing the food recommending that it be kept at or below a specified temperature between 8 ºC and ambient temperatures. 9. 2006 No. It should be noted that the temperature control requirements as detailed in Schedule 4 of the Food Hygiene (England) Regulations 2006 (S. including: Shelf life: where the minimum durability or 'use by' indication is required according to Regulation 20 or 21 of the Food Labelling Regulations 1996 (form of indication of minimum durability and form of indication of ‘use-by date’) the period up to and including that date.
9 Other Relevant Legislation The following regulations about fish designations need to be considered: • Regulation (EC) No 104/2000 on the common organisation of the markets in fishery and aquaculture products Regulation (EC) No 2065/2001 laying down detailed rules for the application of Council Regulation (EC) No 104/2000 as regards informing consumers about fishery and aquaculture products • Note: Requirements for preserved fish are laid down elsewhere. 14. 2076/2005.12.2004.4. 2074/2005 and Regulation (EC) No. 2076/2005. Regulation (EC) No. L226. 1663/2006. Regulation(EC) No.2004. Regulation (EC) No.2004. 1791/2006 and 1021/2008 and as read with EC Directive 2004/41. 1662/2006. 853/2004 is now set out in a Corrigendum (OJ No. Regulation (EC) No. L226. 30.2005. 3) as amended by Regulation (EC) No. L226. The revised text of Regulation (EC) No. Regulation (EC) No. L139.2006. L338. 852/2004 is now set out in a Corrigendum (OJ No. as read with the corrigenda at OJ No. 62) as amended by Regulation (EC) No. namely.6.4. 852/2004 of the European Parliament and of the Council on the hygiene of foodstuffs (OJ No. 25. 83) as amended by Regulation Nos 882/2004. Regulation (EC) No.2004. Commission Regulation (EC) No. 2076/2005. 2074/2005. 2075/2005 and Regulation (EC) No. 1). 206).2004.6.2004. 55). laying down common marketing standards for preserved sardines. L139. The revised text of Regulation (EC) No. 1688/2005. these are not be discussed here. and Regulation (EEC) 1536/92 laying down common marketing standards for preserved tuna and bonito.10. 1. 2074/2005. 30.6. 2076/2005. 25. 30.LEGISLATION 9. 854/2004 of the European Parliament and of the Council laying down specific rules for the organisation of official controls on products of animal origin intended for human consumption (OJ No. 2073/2005 on microbiological criteria for foodstuffs (OJ No. as amended. 1441/2007. Regulation (EEC) 2136/89. 223 . 22) as amended by Regulation (EC) Nos.4.10 Further Reading Regulation (EC) No. 1791/2006 and 1020/2008 and as read with EC Directive 2004/41. 853/2004 of the European Parliament and of the Council laying down specific hygiene rules for food of animal origin (OJ No. 1019/2008 and as read with Regulation 2073/2005. L283. 22. L139. 25. 854/2004 is now set out in a Corrigendum (OJ No. 9. The revised text of Regulation (EC) No. 2074/2005.
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 854/2004.2000. 27 Commission Regulation (EC) No 2076/2005 laying down transitional arrangements for the implementation of Regulations (EC) No 853/2004. 83) Regulation (EC) No 104/2000 on the common organisation of the markets in fishery and aquaculture products (OJ L 17.12.12. (EC) No 854/2004 and (EC) No 882/2004 of the European Parliament and of the Council and amending Regulations (EC) No 853/2004 and (EC) No 854/2004 ( OJ L 338.1. 22.2005. 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 278.2005. as amended by Commission Regulation (EC) No 1022/2008 OJ L 338.2001.FISH AND SEAFOOD Commission Regulation (EC) No 2074/2005 laying down implementing measures for certain products under Regulation (EC) No 853/2004 of the European Parliament and of the Council and for the organisation of official controls under Regulation (EC) No 854/2004 of the European Parliament and of the Council and Regulation (EC)No 882/2004 of the European Parliament and of the Council. 22. 23. 21.10. 6) 224 .
a 10 to 1.0 x 1. 10.1. 10. and 2.1. Some death will occur at this pH if the temperature is 4 °C.4 Heat resistance Aeromonads are sensitive to moderately high temperatures.1.000-fold increase in number is observed after one week in food systems (1).2 Oxygen requirements Facultative anaerobe. under optimal incubation temperatures.3 minutes.5 µm 10. Aeromonas has the potential to grow over a pH range of ca 4 .1 10. At pH 4.3 .45 °C (1). no growth is observed (1). respectively (1). Morphology Gram-negative rods.5. 10.1. PATHOGEN PROFILES 10. and D-values at 45 and 51 °C have been reported as 29.1.10 (3).3 Temperature Aeromonas grows at temperatures of 2 .10. At 5 °C. 225 .5 minutes.1.1 Aeromonas spp.3. The minimum pH value for growth is thought to be pH 5. 0.0 .5.5 pH Under otherwise optimal conditions. The optimum growth temperature is 28 °C (2).
Group I will not grow at temperatures of 10 ºC or less.2.22.2 Clostridium botulinum Seven different types of C. 226 .4 x 1.3 Temperature All strains of C.2. 0. but the minimum temperatures required to inhibit Groups I (proteolytic group) and II (non-proteolytic group) are different. All C. 10. botulinum grow reasonably well in the temperature range of 20 45 ºC. many foods that are not obviously ‘anaerobic’ can provide adequate conditions for growth. E and. but the spores of this organism are more so. All type A strains are proteolytic and type E strains are usually non-proteolytic. at 85 ºC for 5 minutes. botulinum is an obligate anaerobe.0 µm. Types A. 10. 10.2.2.4 Heat resistance The vegetative cells of C. botulinum is prevented. botulinum are known. which may be inactivated by heating at 80 ºC for 20 . F are the types that are responsible for most cases of human botulism (5.5 .7 . botulinum are not particularly heat resistant.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 .1. 10.1 Morphology Gram-positive. or at 90 ºC for a few seconds. botulinum types produce heat labile toxins.FISH AND SEAFOOD 10. but the interior of the food may do so. It is also important to note that the inclusion of oxygen as a packaging gas cannot ensure that growth of C. forming at least seven different toxins.even as low as 3 ºC (7). B. Thus an aerobically packed product may not support the growth of the organism on the surface.5% NaCl (4).2% sodium chloride (NaCl) (1).2. and is sensitive to > 4.30 minutes. spore-forming rods. and groups I and II are those responsible for the botulism cases. but Group II strains are psychrotrophic. to a lesser extent. 6). There are four main groupings of the organism. A to G.2 Oxygen requirements Although C. 10. types B and F can be either.
they may survive mild heat treatments (70 . parcooked meals with pH value > 5. botulinum are produced by type A and proteolytic B strains (D121ºC=ca 0.0 µm.97 for Group II (5). respectively (8.5 pH The minimum pH for the growth of proteolytic and non-proteolytic strains is pH 4. However.9 x 2. 10. The spores of Group II (non-proteolytic/psychotrophic) strains are less heat resistant than Group I strains. These concentrations correspond to limiting aw of 0. safety margins must be introduced to allow for process variability.94 for Group I and 0. In commercial situations.10.0 and aw > 0. vacuum packed.3 10. grow on the surface of foods unless they are vacuum or gas-packed.6 and pH 5. 10. botulinum spores The most heat resistant spores of Group I C. the minimum aw for growth may be lower.is an obligate anaerobe.85 ºC) and their ability to grow at refrigeration temperatures necessitates their control in foods capable of supporting their growth (e. The organism will grow well in the centre of meat or poultry dishes.97) (10. 11)).7 Characteristics of C. botulinum depends on solute.2 Oxygen requirements C. and 5% (w/w) NaCl is necessary to prevent growth of Group II organisms. 0.2.0.21 minutes) (9). 9).2. but under optimum growth conditions 10% (w/w) NaCl is required to prevent growth of Group I. 10.3. These values have been established under carefully controlled laboratory conditions.3. Note also that if humectants other than salt are used. pH and temperature. therefore.PATHOGEN PROFILES 10.g.6 Aw/ Sodium chloride The minimum water activity (aw) for growth of C.1.0 . 10. particularly by cooking. perfringens .1 Clostridium perfringens Morphology Gram-positive spore-forming rods. 227 .2.3 . where oxygen levels are reduced. It will not.like other clostridia .
and their heat resistance is also affected by the heating substrate. perfringens in relation to food safety is the organism’s ability to grow extremely rapidly at high temperatures. 15.3.97 (glycerol and sucrose respectively) depending on the solute used to control aw (13. death of the vegetative cells of this organism usually occurs rapidly above this temperature (12.93 and 0. the enterotoxin is not heat resistant . 16). 13).8% inhibit growth of most C. perfringens is not a tolerant organism with respect to pH. although prior growth at high temperatures or the presence of fat in a food will result in increased heat resistance. In addition. depending on strain and other conditions. type of solute. Under otherwise ideal conditions. 10.3.it is destroyed by heating at 60 ºC for 10 minutes (14. perfringens has the potential ability to grow within the temperature range 15 . 18). 13).FISH AND SEAFOOD 10.5 pH C. The D-value for C.3. (It is unusual for spores to be formed in foods after the growth of this organism (14)). As in the case of other factors limiting the growth/survival of this organism. perfringens spores The spores of C. the limits for aw are affected by temperature. While some growth can occur at 50 ºC. Salt concentrations of 6 .45 ºC. although C. perfringens strains.50 ºC.3.7 Characteristics of C. perfringens appears to be 0. Spores. 10.4 Heat resistance Exposure to a temperature of 60 ºC or more will result in the death of vegetative cells of C.6 Aw/Sodium chloride C.5 minutes (17). It grows best at pH values between 6 and 7 (the same pH as most meats). Its optimum temperature for growth is 43 . 10. Recorded heat 228 .3. however. perfringens can vary quite considerably in their heat resistance. lower concentrations may be effective in combination with other factors. perfringens is not tolerant of low water activities. perfringens. very limited growth may occur at pH values over the range pH ≤ 5 and ≥ 8. At cold temperatures.3 Temperature The most significant characteristic of C. perfringens in vacuum packed beef (18). pH. will survive greater extremes of pH (and aw) (12. 0 . perfringens in roast beef at 60 ºC is 14.3. etc. vegetative cells die rapidly (12). 10. Some studies indicate that the presence of 3% NaCl delays growth of C.10 ºC. The lowest aw recorded to support the growth of C.
is between 30 and 37 ºC.23. growth at low temperatures can be very slow. In foods.5 x 0.19 minutes.0. monocytogenes is reported to be 45 ºC (20). pH < 5. D-values for L. -0.3 . however. 10.4 10.4.1 Listeria spp.98 and 0.3 is the lowest value where growth has been recorded.4. or even below 0 ºC. and where acetic acid is used as an acidulant. Morphology Gram-positive short rods. Its optimum growth temperature.5 . and temperature. The upper temperature limit for the growth of L. it is not a sporeformer.4 . using hydrochloric acid as an acidulant).64.0 µm. 10.4.2 Oxygen requirements Aerobe or microaerophilic.4 ºC is probably a more likely minimum for growth in foods (19). 229 .0 minutes for heat resistant spores to 1. requiring days or weeks to reach maximum numbers.3 Temperature Listeria monocytogenes is unusual amongst foodborne pathogens in that it is psychrotrophic. Under ideal conditions.4 Heat resistance L. 10. 0. However.4. respectively (21).8 minutes for heat sensitive spores (12). 10. monocytogenes is not a particularly heat resistant organism. however. the organism is able to grow at pH values well below pH 5 (pH 4.albeit slowly .2 has been suggested as the lowest working limit (22). monocytogenes in crawfish tail meat at 55.2. It has been reported to have slightly greater heat resistance than certain other foodborne pathogens.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.PATHOGEN PROFILES resistance values at 95 ºC (D-values) range from 17. the lowest limit for growth is likely to be considerably higher .6 . 10.at refrigeration temperatures down to.especially at refrigeration temperatures. 1.2. so can be destroyed by pasteurisation. being potentially capable of growing . 60 and 65 ºC were reported as 10.4.
5 10. 10. 10.8. 25). peritrichous flagella.90.0 .3.8 .0 x 1 .6.7 x 1.5. or even grow.97. 10.5.0 µm.1.5 (24.45 ºC.5 .5 pH Plesiomonas has a pH range for growth of pH 4.5. The bacterium may survive for long periods at aw values as low as 0.5 .4.1 Salmonella spp.0. Morphology Gram-negative short rods. 10. 10.6 Aw/Sodium chloride Plesiomonas can grow well at NaCl concentrations up to 4% (24.5.12% NaCl or more). at salt levels found in foods (10 . 230 .1 Plesiomonas Morphology Gram-negative.2 Oxygen requirements Facultative anaerobe.6 10. the optimum being 30 ºC (23). 0. It is likely to survive.5. monocytogenes is quite tolerant of high NaCl/low water activities.3 µm.83 (20).3 Temperature Plesiomonas can grow in the temperature range of 8 . but has been shown to be able to grow at an aw level of 0. 0.5. 10. It grows best at aw of ≥ 0. 25).4 Heat resistance Pasteurisation at 60 ºC for 30 minutes has been reported as being effective in killing Plesiomonas shigelloides (24).6 Aw/Sodium chloride L. 10.FISH AND SEAFOOD 10.
10. Salmonellae are quite resistant to freezing.37 ºC.6.20 times more heat resistant than the average strain of Salmonella at high aw (27). high fat or low moisture levels (low aw) will reduce the effectiveness of heat treatments.5 ºC.48 ºC. Salmonella senftenberg 775W is about 10 . The growth of Salmonella is generally inhibited by the presence of 3 .93 (as reported in dried meat and dehydrated soup). depending on the type of acid and temperature. However.8 .3 Temperature Salmonellae can grow in the temperature range of 7 . substrate.5 .4% NaCl.6.6 Aw/Sodium chloride Where all other conditions are favourable. D60 ºC values normally range from about 1 to 10 minutes. and with an appropriate acid. although salt tolerance increases with increasing temperature (28).5. 10. or possibly 0. under otherwise ideal conditions.0. a heat resistant organism. Some death will occur at pH values of less than about 4.6.4 Heat resistance Salmonella is not a spore-forming organism.9. Furthermore. some strains are able to grow at temperatures as low as 4 °C (26).945. therefore. pasteurisation and equivalent treatments will destroy the organism under normal circumstances. 10. 231 . the optimum being 35 . strains vary in their ability to withstand heating. However.6. and appropriate heat treatments must be determined experimentally for low-aw foods.7.PATHOGEN PROFILES 10.5 pH Salmonella has a pH range for growth of pH 3. temperature and pH. Salmonella has the potential to grow at aw levels as low as 0. depending on serotype.6.2 Oxygen requirements Facultative anaerobe. The optimal pH for Salmonella growth is between 6. It is not. 10.5. Growth is slow at temperatures below about 10 ºC. Salmonellae are quite resistant to drying. with a z-value of 4 .
Under most circumstances.7. Most strains of staphylococci can grow within the pH range 4. In meat macerate. etc. the D60 ºC value is 2 .). 10.7 . 10. under otherwise ideal conditions (29.0. NB: The growth of Staph.2 to 9. aureus viability.9 µm diameter. with an optimum of 35 .37 ºC (29). 10. but may cause some cell damage (30).7 10. Staph.1 Staphylococcus aureus Morphology Gram-positive cocci.5). The following relate to limits for growth only. 0.7.4 Heat resistance Heat resistance depends very much on the food type in which the organism is being heated (conditions relating to pH. Freezing and thawing have little effect on Staph.7.2 Oxygen requirements Facultative anaerobe.7. 31). It can survive well at low temperatures. aw and temperature (sensitivity to acid increases with temperature). stressed cells can also be less tolerant of heating.3 (optimum 7.20 minutes. 232 . The limits for toxin production are also narrower than for growth.3 Temperature Under otherwise ideal conditions.0 .5 ºC. As is the case with other bacteria. fat content.7. however.FISH AND SEAFOOD 10. aureus is more limited under anaerobic than aerobic conditions. aureus can grow within the temperature range 7 – 48. aw. the organism is heat sensitive and will be destroyed by pasteurisation.5 pH The pH at which a staphylococcal strain will grow is dependent upon the type of acid (acetic acid is more effective at destroying Staph. aureus than citric acid). 10. depending on aw.7.
2.4 .7.86 is the generally recognised minimum in foods (32).6 Aw/Sodium chloride Staph. pH.7.8 10.2 . In general. aureus can grow in 7 .0 . However. 233 .1 Vibrio cholerae Morphology Gram-negative curved rods.0.>0.5) (29.5 . Staph.8. It can grow over the aw range 0.43 ºC (35). 31) between 0. In general.10% NaCl.PATHOGEN PROFILES 10. 10.2 Oxygen requirements Facultative anaerobe. * dependent on temperature.8. atmosphere.46 ºC (optimum between 35 and 40 ºC) (very little toxin is produced at the upper and lower extremes) (31) 5. An effect of increasing salt concentration is to raise the minimum pH of growth. aureus is unusual amongst food-poisoning organisms in its ability to tolerate low water activities. strain. especially vacuum packed foods (33) Temperature: pH: *Aw: Atmosphere: Heat Resistance: Enterotoxins are quite heat resistant.87 and > 0.7 Limits permitting toxin production 10 .83 . an aw of 0. Staphylococci are more resistant to salt present in foods than other organisms. but certain strains can grow in 20% NaCl.99 little or no toxin production in anaerobically packed foods. 10.6 µm 10.7. 0. with a temperature range for growth of 10 .8 x 1.3 Temperature The optimum temperature for growth has been reported as 30 ºC. 34).8.9. 10. heating at 100 °C for at least 30 minutes may be required to destroy unpurified toxin (31. and solute.0 (optimum 7.99 aerobically under otherwise optimal conditions.
cholerae is sensitive to heat at temperatures of >45 ºC.5 ºC).9.0.9. and it is sensitive to refrigeration temperatures (0 . 10.1 . but not acid conditions. The D-value for V. parahaemolyticus is easily destroyed by drying.984 (35). declining in number during storage.97 . 10. cholerae may persist on frozen fish for >180 days (36). are 5 and 43 ºC. respectively.0% NaCl (35).8. with optimum at 7.1 Vibrio parahaemolyticus Morphology Gram-negative curved rods. there is evidence that V.6 (35. 10.4. 38). It can grow in the presence of 0. 234 . 10.3 Temperature The minimum and maximum temperatures reported for growth in laboratory media. 37.6 Aw/Sodium chloride V. but only moderately sensitive to freezing (40). with an optimum of 37 ºC (39).9. with an optimum of 0.8. cholerae can grow over the aw range 0. 10. 10. it has a pH range for growth of pH 5 .4 Heat resistance V.5 pH This organism is tolerant of high pH values.9.9 10. V. under otherwise favourable conditions.2 Oxygen requirements Facultative anaerobe.8. cholerae in crab meat homogenate at 60 ºC is 2.998.65 minutes (35).FISH AND SEAFOOD Also.6.
10. and are readily destroyed by cooking.5% NaCl. 10.29 minutes. parahaemolyticus is unable to grow in the absence of salt.8. with a temperature range for growth of 8 . parahaemolyticus will grow at aw levels as low as 0. and 2. but the optimum concentration is 2 .10. 10.9. 41). 10.4 Heat resistance V.02 .6 Aw/Sodium chloride V. The D-values at 55 ºC for clam and crab homogenate are 0.43 ºC (42).PATHOGEN PROFILES 10. 10.10.4 Heat resistance V. 41). The organism can survive for long periods at freezer temperatures (42).10 Vibrio vulnificus 10. depending on solute. 235 .6 (41).922.11.9. with an optimum pH of 7.4% (39.8 . parahaemolyticus are not heat resistant.5 pH The pH range for growth is 4.5 minutes.3 Temperature The optimum temperature for growth has been reported at 37 ºC. respectively. The optimum concentration of NaCl equates to aw of approximately 0. its upper and lower limits for growth are 8 (although some reports suggest 10) and 0. it has been reported that cooking oysters for 10 minutes at 50 ºC should ensure destruction of the organism (43).6 .0. 10. vulnificus is heat sensitive.9. respectively (39).1 Morphology Gram-negative curved rods.2 Oxygen requirements Facultative anaerobe. 10.10. V.98 (35.
9. Eds. 1989. Hauschild A. Clostridium botulinum and Clostridium perfringens.M.O.C. Washington D.. Microbiology and Molecular Biology. Aeromonas hydrophila group. 10. 121-76.L. 111-89.P.. Ed. in Foodborne Pathogens.P. ASM Press. 279-303. Hui Y. Cliver D. Dodds K.M.. Johnson E.H. 97-131.R. 2005. New York.W. Clostridium botulinum. vulnificus grows over the range 0.11 1..10. 383-408. 1994. Hauschild A.. Montville T. Potential hazards associated with REPFEDS.H. Dodds K..L. Bhunia A.K. Beuchat L. Kim J. Volume 1.D. Smith J. Doyle M.O. Marcel Dekker. Growth and heat resistance of psychrotrophic Clostridium botulinum in relation to ‘sous vide’. with an optimum of 2.. with an optimum pH of 7. 1-27.. Ed. Doyle M. Novak J. London. Marcel Dekker Inc. 8.. Miliotis M. New York.0% NaCl.W.. Eds.A. Marcel Dekker. Doyle M. 1995. in Foodborne Bacterial Pathogens. Betts G.L.5 . Abeyta C. 1993. Hauschild A. International Commission on Microbiological Specifications for Foods. Lund B. Eds..W.5% (42). Gorham J... Juneja V. in Clostridium botulinum: Ecology and Control in Foods. 1989. Volume 1: Diseases Caused by Bacteria. Caister Academic Press. Faris A. Cliver D. Fratamico P.H.. in Foodborne Bacterial Pathogens. Ed. in Food Microbiology: Fundamentals and Frontiers. New York. 6 (1).. Palumbo S.FISH AND SEAFOOD 10. Volume 5: Microbiological Specifications of Food Pathogens. 1996. Eds. Food Control.D.H.5. Stelma G..W.. 10. Aeromonas. 10. 6. 2. 2003.D. 11. Notermans S. in International Handbook of Foodborne Pathogens.. New York. New York. Marcel Dekker. 1994. in Foodborne Disease Handbook.. 1-19.P.L.M. Hui Y.N. 57-63. Gorman J. Dodds K. Eds. Eds. 4. Diseases Caused by Bacteria. 236 .D. 357-67.5 pH The pH range for growth is 5 . Aeromonas hydrophila. Austin J. Marcel Dekker. in Foodborne Disease Handbook. New York. Peck M.. Kovacek K. 5-19.R. in Microorganisms in Foods. Blackie..E.8 (42). in Clostridium botulinum: Ecology and Control in Foods. Great Britain.H. 5.R. Clostridium botulinum.W. 7.6 Aw/Sodium chloride V. Bier J.10. New York. Marcel Dekker. Stelma G. 1993. 3..J. Marcel Dekker.. References International Commission on Microbiological Specifications for Foods. Foegeding P. Principles of control. Murrell K. 329-49. Murrell K. Aeromonas species. Eds.10. Gaze J. 2001.N. Clostridium botulinum.H.
Foodborne disease due to Bacillus and Clostridium species. 2007. Nagaraja K. 23. Evans M.P. in Foodborne Pathogens: An Illustrated Text. 1990. ASM Press. Effect of time and temperature on growth of Salmonella enteritidis in experimentally inoculated eggs. Hui Y. Halvorson D. Ed. Ed. in Foodborne Diseases. 982-6. Volume 5: Microbiological Specifications of Food Pathogens.H. 129-38. Wolfe Publishing Ltd. in Foodborne Infection and Intoxication. International Commission on Microbiological Specifications for Foods.. Doyle M.R. 20. 1994.O.J. 2006. 1989. 1989. Listeria. 1996. Clostridium perfringens. London. Washington D. 137-64. Marcel Dekker. Emery D. Clostridium perfringens food poisoning. Growth of Listeria monocytogenes at refrigeration temperatures. Ryser E.G. Labbe R.. 17. Volume 5: Microbiological Specifications of Food Pathogens.J. Clostridium perfringens. Avian Disease..O. 1996. Lund B. in Food Microbiology: Fundamentals and Frontiers. Eds. Academic Press. Clostridium perfringens. London.H. London. 133-67. New York. Listeria monocytogenes.. McClane B..A. Marth E. Elsevier. Matthews K. Evans M. Clostridium perfringens. 22. in Foodborne Bacterial Pathogens.M...J.An Introduction.O... 1989. Archer P. Murrell K. Volume 5: Microbiological Specifications of Food Pathogens. Wrigley D.A. London. International Commission on Microbiological Specifications for Foods. Rinke H. 112-25.A. 1991. 2005. in Microorganisms in Foods.G.K. Marcel Dekker. 201-8. Ed. 26.V. Varnam A. Montville T. 24. 25. Plesiomonas. 208-13. Ed. International Commission on Microbiological Specifications for Foods. 191-243. Beuchat L. 16.M. Varnam A. 2001. ASM Press. Blackie..P. 229-40. London. International Commission on Microbiological Specifications for Foods.D. Journal of Applied Bacteriology. Plesiomonas shigelloides. 1996.. Eds.A. Kim C.T. 351-82. Marcel Dekker. 157-62. Listeria monocytogenes. Montville T. International Commission on Microbiological Specifications for Foods. 68 (2). Marcel Dekker... 21. 141-82. New York.PATHOGEN PROFILES 12. 336 (8721). 311-25. Plesiomonas shigelloides. Lancet. International Commission on Microbiological Specifications for Foods. Riemann H. Juneja V.. Gorham J. Doyle M. 735-42.G. Eds. Clostridium perfringens gastroenteritis. 237 . 19. 1990. in Food Microbiology .J. New York. 18.P..R. 15.H. Ed. in Foodborne Disease Handbook. London. Blackie. 1990. Doyle M.P. Banks J. Volume 1. Listeriosis and Food Safety. in Microorganisms in Foods. Diseases Caused by Bacteria.C.A. Cliver D. Cliver D. in Microorganisms in Foods. Eds. Ed. Eds. 14. 33. Labbe R. Blackie.H. New York. Johnson E.R. 13. Cliver D... Washington D. in Foodborne Bacterial Pathogens.C. Walker S. Koburger J..
Jay J. Microbiological Specifications of Food Pathogens. Salmonella. 31. International Commission on Microbiological Specifications for Foods. D’Aoust J-Y. 2005. Warner R.. CDR Review. Staphylococcal Food Poisoning. New York.K.M.. Doyle M. 1996. 4 (13).P.A. 34. 33.P.C. 238 . Academic Press. London. Lee-Wong A. Eds.C. Marcel Dekker. Ed. in Modern Food Microbiology. 37. New York. 985-98. 217-64.L. 28. Cliver D. On the resisting power of Vibrio cholerae and typhoid Bacillus against low temperatures. 85-106. Cleland D. 8. Staphylococcus aureus as a food pathogen: staphylococcal enterotoxins and stress response systems. Jay J. Doyle M. London. 35. Smith J.J. Foodborne illness (Part 1): Staphylococcal (“Staph”) food poisoning. London. 1994. Wilkinson B.D. 545-66. Stress Response and Resistance. Cliver D. 2005.P. Volume 5. Ed. International Commission on Microbiological Specifications for Foods. Marcel Dekker.H. Staphylococcal gastroenteritis.A. 1983. in Foodborne Pathogens: Microbiology and Molecular Biology. 13 (11). 273-84. 523. 1993. Food and Environmental Sanitation. Academic Press. Golden D. 642. in Microorganisms in Foods. Dairy..S. 36. Reed G. 426-35. 1996. 327-445. International Commission on Microbiological Specifications for Foods. Riemann H.M. Staphylococcal intoxications. 1989.S. Experimental investigations concerning the prevention of epidemic diseases due to aquatic products (Fourth Report).FISH AND SEAFOOD 27. London. International Commission on Microbiological Specifications for Foods.O. Ed.R. 1930. 45 (3).J. Salmonellae. Loessner M. New York. Ed. Eds. 2005. Blackie. Bergdoll M. Vibrio parahaemolyticus. 327-52. in Foodborne Pathogens. International Commission on Microbiological Specifications for Foods. Crowcroft N. 32. Japanese Journal of Experimental Medicine. 38. Volume 5.. in Foodborne Infections and Intoxications. 1989. Oliver J.. Eds. Loessner M.. R15764.S. 1990. Vibrio cholerae. London. Wymondham. 1996. 29. 543-68.. Ed. in Microorganisms in Foods. Bergdoll M. Blackie. Woodhead Publishing Ltd. Springer Science. Tohyama Y. Staphylococcus aureus.O. Fratamico P. Distribution of Vibrio vulnificus and other lactose-fermenting vibrios in the marine environment. in Understanding Pathogen Behaviour Virulence. Volume 5: Microbiological Specifications of Food Pathogens. Ed. in Microorganisms in Foods. Blackie. in Foodborne Disease. 414-25.M. Twedt R. Vibrio parahaemolyticus. Cholera: current epidemiology. Caister Academic Press. Applied and Environmental Microbiology. 2005. Bhunia A.. 30..A. Ed. 331-57. 40. Griffiths M. in Foodborne Bacterial Pathogens. Stewart G.M. Cambridge. Golden D. Microbiological Specifications of Food Pathogens. International Commission on Microbiological Specifications for Foods.62. Gustafson J.. 39.
55 (12).M. Yersinia.M. 985-9.W.J. Cold storage and mild heat treatment as processing aids to reduce the numbers of Vibrio vulnificus in raw oysters.D.. 43. 657-78. International Commission on Microbiological Specifications for Foods. Jay J..PATHOGEN PROFILES 41.J. London. in Modern Food Microbiology. Ruple A. Foodborne gastroenteritis caused by Vibrio. Loessner M.A. New York. 1996. 436-9.. Journal of Food Protection.. Golden D. and Campylobacter species.A. Loessner M. Springer Science. International Commission on Microbiological Specifications for Foods. Cook D. Microbiological Specifications of Food Pathogens. 1992. 2005. Volume 5. Vibrio vulnificus. Golden D.. Blackie. Jay J. 239 . Ed. in Microorganisms in Foods. 42.
FISH AND SEAFOOD 240 .
gov.org.seafish.gov.uk or v.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.org/ .org.gov.scotland.uk/ Department for Environment Food and Rural Affairs (Defra) Nobel House 17 Smith Square London SW1P 3JR United Kingdom Tel: + 44 (0) 207 2386000 Fax: + 44 (0) 207 2382188 241 Email: email@example.com 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 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: info@ifm.CONTACTS Addresses of Trade Associations and Professional Bodies Association of British Salted Fish Curers and Exporters c/o Cawoods (Fish Curers) Ltd Estate Rd No 6 South Humberside Industrial Estate Grimsby North East Lincolnshire DN31 2TG United Kingdom Tel: + 44 (0) 1472 342248 Fax: + 44 (0) 1472 353100 Email: email@example.com Web site: firstname.lastname@example.org Web site: www.uk Web site: www.gsi.ifm.
is/ 242 . Baelskaai 25 8400 Oostende Belgium Tel: + 32 59321876 Fax: + 32 59322840 Email: info@eapo.FISH AND SEAFOOD Department of Agriculture and Rural Development.asp Food and Agriculture Organization of the United Nations Fisheries and Aquaculture Department Viale delle Terme di Caracalla 00153 Rome Italy Tel: + 39 (06) 57051 Fax: + 39 (06) 57053152 Email: FAO-HQ@fao.dardni.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: difta@difta. Dublin Eire Ireland Tel: + 353 1 2144100 Web site: www.gov.fao.is Web site: www.bim.no Web site: www.ie/templates/homepage.13 Postbox 6122 N-9291 Tromsø Norway Tel: + 47 77 629000 Fax: + 47 77 629100 Email: email@example.com Web site: www.fiskehandlerne.no/ Matís ohf (Icelandic Fisheries Laboratories) Skúlagata 4 101 Reykjavík Iceland Tel: + 354 422 5000 Fax: + 354 422 5001 Email: firstname.lastname@example.org/s Norwegian Institute of Fisheries and Aquaculture Ltd Muninbakken 9 .dk Danmarks Fiskehandlere H.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.gov.eapo.C.suite. 1 1553 København V Denmark Tel: + 45 35 372023 Fax: + 45 35 372033 Email: Info@fiskehandlerne.com Web site: www.net Irish Sea Fisheries Board PO Box 12 Crofton Road Dun Laoghaire Co.fiskeriforskning.org/fishery/ European Association of Fish Producers’ Organisations H.email@example.com Web site: www.dk Web site: www. Andersens Boulevard 37.uk/index/fisheriesfarming-and-food.org Web site: www. Northern Ireland Fisheries and Rural Policy Division 4th Floor North Dundonald House Stormont Estate Belfast BT4 3SB United Kingdom Tel: + 44 (0) 28 90524999/ 4991 Fax: + 44 (0) 28 90378323 Email: dardhelpline@dardni.
uk/en/index.ifst.fdf.gov.htm Food and Drink Federation (FDF) 6 Catherine Street London WC2B SJJ United Kingdom TeI: + 44 (0) 207 8362460 Fax: + 44 (0) 207 8360580 Email: firstname.lastname@example.org Web site: www.csiro.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.co.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 Institute of Food Science and Technology (IFST) 5 Cambridge Court 210 Shepherds Bush Road London W6 7NL United Kingdom TeI: + 44 (0) 207 6036316 Fax: + 44 (0) 207 6029936 Email: firstname.lastname@example.org 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.org Web site: www.au Web site: www.cmar.dh.gov.seafood.au New Zealand Seafood Industry Council Private Bag 24-901 Wellington 6142 New Zealand Tel: + 64 4 3854005 Fax: + 64 4 3852727 Email: email@example.com/ 243 .co.uk Department of Health Richmond House 79 Whitehall London SW1A 2NS United Kingdom Tel: + 44 (0) 207 2104850 Email: firstname.lastname@example.org CSIRO Marine Laboratories Castray Esplanade GPO Box 1538 Hobart Tasmania 7001 Australia Tel: + 61 3 62325222 Fax: + 61 3 62325000 Email: email@example.com Other Sources of Information British Standards Institution 389 Chiswick High Road London W4 4AL United Kingdom Tel: + 44 (0) 208 9969001 Fax: + 44 (0) 208 9967001 Email: firstname.lastname@example.org Web site: www.uk Web site: www.
1211 Geneva 27 Switzerland Tel: + 41 22 7912111 Fax: + 41 22 7913111 Email: email@example.com 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/ SIK .uk/index.uk/ Health Protection Scotland Clifton House Clifton Place Glasgow G3 7LN United Kingdom Tel: + 44 (0)141 3001100 Fax: + 44 (0)141 3001170 Email: hpsenquiries@hps.FISH AND SEAFOOD Society for Applied Microbiology Bedford Heights Brickhill Drive Bedford MK41 7PH United Kingdom Tel: + 44 (0) 1234 326661 Fax: + 44 (0) 1234 326678 Web site: www.hps.int/foodsafety World Health Organization (WHO) Headquarters CH .net/web/indexen.int Web site: http://www.who.euro.uk Web site: www.who.nhs.euro.se Web site: www.sofht. jsp Food Safety 10 .int Web site: www.int/en/ WHO Regional Office for Europe Scherfigsvej 8 DK-2100 Copenhagen Ø Denmark Tel: + 45 39 171717 Fax: + 45 39 171818 Email: email@example.com. Web site: www.The Swedish Institute for Food and Biotechnology Box 5401 SE-402 29 Göteborg Sweden Tel: + 46 31 3355600 Fax: + 46 31 833782 Email: firstname.lastname@example.org/ FAO/WHO Collaborating Centre for Research and Training in Food Hygiene and Zoonoses Robert von Ostertag Institute Postfach 330013 D-14191 Berlin Germany Tel: + 49 (0) 188 84122155/ 2157 Fax: + 49 (0) 188 84122957 Email: email@example.com Rome Italy Tel: + 39 06 487751 Fax: + 39 06 4877599 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.org.uk/scieh.scot.uk Web site: www.de 244 .org Web site: www.nhs.
au/ International Association for Food Protection http://www.int/foodsafety/en/ European Commission: Activities of the European Union food safety http://europa.csiro.uk/ 245 .fpa-food.fsai.who.org/ Royal Institute of Public Health (UK) http://www. Beverage and Consumer Products Companies http://www.org/cms/ Useful Web Sites Gateway to Government food safety information (US) http://www.gov/foodsafety/ Food Safety Authority of Ireland http://www.cdc.org. Van Buren Suite 1000 Chicago IL 60607 United States of America Tel: + 1 3127828424 or + 1 8004383663 Fax: + 1 3127828348 Email: email@example.com.FoodSafety.eu/pol/food/index_en.ie/ Food Science Australia http://www.htm Centre for Disease Control and Prevention (CDC) (US) http://www.org Web site: http://www.CONTACTS US FDA .sofht.org/ The Association of Food.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.co.riph.gov/ World Health Organization (WHO): Food safety programmes and projects http://www.org/ Institute of Food Technologists http://www.fda.ift.uk/ Society of Food Hygiene and Technology (UK) http://www.ift.foodprotection.
FISH AND SEAFOOD 246 .
111 Astroviruses. as autochthonous bacterial flora of fish 3 as spoilage organisms in oysters 65 as spoilage organisms in temperate crustacean shellfish 80 as spoilage organisms in tropical crustacean shellfish 81 in molluscan shellfish 60 Aerobic plate counts. growth and survival in chilled and frozen raw fish 19 of concern in preserved fish 115 Aspergillus spp. preserved by dry/wet salting 99 Anisakis spp. as TTX producers 145 Amnesic shellfish poisoning (ASP) toxins 146-8 caused by domoic acid in crustacean shellfish 87 Anchovies. pathogenic microorganisms in molluscan shellfish 59-60 as contaminant in enrobing process 37 as risk in breaded shrimp and prawn products 82 growth and survival in chilled and frozen prepared fish products 45 growth in vegetable and rice 247 .INDEX Achromobacter spp. source of PSP toxins 142 Alexandrium ostenfeldii. breakdown of – as indicator of fish freshness 10-1 Azaspiracids 158-9 detection methods 160 origins and distribution 159 toxicology 159-60 Bacalao 97. growth and survival in crustacean shellfish 86 Aeromonas sobria. causing spoilage of dried fish 110. 69-70 Aeromonas hydrophila. as autochthonous bacterial flora of fish 3 in fish sauce 127 Bacillus cereus. in crustacean shellfish 86-7 ATP. to test microbiological criteria 19 Aeromonadaceae. as autochthonous bacterial flora of fish 3 Aeromonas spp. as spoilage organism in oysters 65 as spoilage organism in tropical crustacean shellfish 81 in molluscan shellfish 60 Alexandrium. in crustacean shellfish 87 Alteromonas spp. 102 Bacillus spp. growth and survival in crustacean shellfish 86 Air drying 101 Alcaligenes. 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. planktonic source of spirolides in shellfish 163 Allergens.
affected by azaspiracids 159 affected by cyclic imines 161 Botulism. of fresh fish spoilage 11 Chemical pollutants. growth and survival in crustacean shellfish 86 growth and survival in molluscan shellfish 59-60. caused by Vibrio cholerae O1 17 Ciguatoxins 155-8 detection methods 157-8 origins and distribution 1567 toxicology 157 Clams. description and uses 56-7 effects of processing on microflora 60-4 hot-smoking 107 Clear smoke treatments 108 Clostridium. as autochthonous bacterial flora of fish 3 Clostridium botulinum. 137. concern with respect to use of MAP 7 growth and survival in chilled and frozen prepared fish products 45 growth and survival in chilled and frozen raw fish 15-6 growth and survival in crustacean shellfish 84 growth and survival in cured. of fish – effects on microflora 5-6 Carbohydrate. 98-9 Brochothrix thermosphacta. 113. associated with consumption of fish/fish products 16. 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 . as gas in modifiedatmosphere packaging 6 CCP Decision Tree (in HACCP plan) (Fig) 185 Ceviche. cause of neurotoxin shellfish poisoning 153-5 origins and distribution 1545 toxicology 155 importance in wet salting of fish 96 Brining 93. preserved by marinating 100 Chemical detection methods. in crustacean shellfish 86-7 Campylobacter coli. mould causing spoilage of dried fish 110 Battering process 36-8 Bivalves. 136. in crustacean shellfish 87 Chilled prepared fish products 27-51 Chilled raw fish 1-25 Chilled storage. 131 Carbon dioxide. of recipe fish meals 43 Cholera. indigenous – in fish (Table) 4 non-indigenous – in fish (Table) 4 Bakasang. fish sauce 123 Basipetospora halophila. 226 Breading process 36-8 Brevetoxins. growth and survival in crustacean shellfish 86 pathogenic microorganisms in molluscan shellfish 59-60 Campylobacter jejuni.FISH AND SEAFOOD components of seafood ready meals 40 Bacterial pathogens. importance in lactic acid fermentation in fish products 130. for PSP toxins 144 Chemical indicators. as cause of spoilage of MAP fish 15 Calciviruses. 68-9 Capture and handling.
149-50. 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. 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. of fish – analysis of 10 Diarrhetic shellfish poisoning (DSP) toxins 148-51 detection methods 150-1 origins and distribution 14950 toxicology 150 Dinophysis spp. as contaminants in scampi 36. cooked – effects of processing on microflora 83 forms for purchase 80 freshness indicators 83 Crayfish. adverse condition on poorly preserved fish 110 EC grading scheme. 106 Contacts 241-5 Conway method. growth and survival in crabmeat 84 growth and survival in cured. smoked and dried fish 114 pathogenic microorganisms in molluscan shellfish 59-60 249 . as spoilage organisms in tropical crustacean shellfish 81 Coryneforms. as risk in breaded shrimp and prawn products 82 growth and survival in chilled and frozen prepared fish products 45-6 pathogen profile 227-9 Coated fish products. 152 Diphyllobothrium.INDEX pathogen profile 226-7 Clostridium perfringens. processing 334 Cockles. for detection of DA 148 ELISA kit (DSP-Check). to determine fresh fish spoilage 11 Corynebacteria. forms for purchase 80 Crustacean shellfish 79-92 Cured fish 93-121 Cyclic imines 160-1 detection methods 162 origins and distribution 161 toxicology 162 Degradation. as autochthonous bacterial flora of fish 3 Crabs. of prepared fish products 44 Escherichia coli. growth and survival in chilled and frozen raw fish 19 of concern in cold-smoked salmon 115 Domoic acid (DA). source of azaspiracids 159 source of DSP toxins 148-9. hot-smoking 107 ELISA assay. to determine fresh fish spoilage 11 Eels. to detect NSP toxins 155 ELISA kit (Biosense). description and uses 59 effects of processing on microflora 60-4 Cold-smoking 104-5. to detect DSP toxins 150-1 Enrobing process 36-8 Enterobacteriaceae. smoked and dried fish 114 Enzymic spoilage. of fish – different processes 94 Dun.
as source of inulin in lactic acid fermentation in fish products 134 Garos. as source of ciguatoxins 156 Garlic. definition 1 Fermented fish 123-40 Fermented fish/salt products of Thailand (Table) 125 Fish and shellfish toxins 141-74 Fish cakes. effects of processing 127-30 Fish sauces and pastes of south-east Asia (Table) 124 Fish/salt/carbohydrate products of Asia (Table) 125. to measure fish skin resistance 10 Flavobacterium. fish sauce 123 HACCP.FISH AND SEAFOOD EU legislation. of fish – effects on microflora 5-6 Hepatitis (non-A. source of PSP toxins 142 Gyoshoyu. 130-5 Fish-spoilage bacteria 12 Fishtester. of fresh fish spoilage 911 Evisceration. of chilled and frozen prepared fish products – preparation 33-6 Fish sauces 123-4 Fish sauces and pastes. preserved by marinating 101 Gravlaks. concerning crustacean shellfish 87-8 European Grading Scheme. causing pink condition on fish surface 110 in fish sauce 127 Halococcus spp. for evaluation of fish quality 9 Eurotium. fermented fish product 124 General Pathogen Surveillance Programme 84 Gravad fish. causing pink condition on fish surface 110 in fish sauce 127 Handling. mould causing spoilage of dried fish 110 Evaluation. preserved by marinating 100 preserved by dry/wet salting 99 Histamine fish poisoning. in crustacean shellfish 86-7 in molluscan shellfish 59-60 Hepatitis A virus. fermented fish product 126 Green cured split cod 97. caused by Photobacterium phosphoreum 15 Hon kare bushi 107 Hot-smoking 105-7 250 . specific EU hygiene rules 196-224 Fresh fish. of prepared fish products 44 Gambiertoxins. member of cyclic imine group 161 Gymnodinium catenatum. spoilage 9-11 Freshness grading schemes 9 Frozen prepared fish products 27-51 Frozen raw fish 1-25 Frozen storage. 102 Gymnodimines. in crustacean shellfish 86-7 in molluscan shellfish 59-60 Herring. of fish at sea 5 Fatty fish. non-B) viruses. processing 35 Fish component. 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. 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.
outbreak caused by gravid 115 Lobster. cooked – effects of processing on microflora 83 forms for purchase 80 freshness indicators 83 Mackerel. food hygiene 189-224 Lingulodinium polyedrum. preserved by salt-boiling 100 Ionising radiation. concerning crustacean shellfis 88 Jellied eels.INDEX Hygiene regulations for foodstuffs production and distribution (EU) 190-6 Ice storage. as source of botulism 136 International legislation. initial – of chilled and frozen prepared fish products 31-2 of chilled and frozen raw fish 3-5 251 . for microbial inactivation in fish products 127. in fish fillet 1 Lipid oxidation. preserved by marinating 101 Microbiological criteria. 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. preserved by salt-boiling 100 Karenia brevis. 135 I-zushi. smoked and dried fish 114-5 hazard in cold-smoking 105 hazard in mild-processed marinated products 101 Listeriosis. as source of yessotoxins 152 Lipid content. 131 Lactic fermented fish products (Table) 126 fish species used in the Philippines (Table) 126 Lactobacillus. producer of brevetoxins 154 Katsuobushi 107 Kench curing 97-8 Klipfish 97 Lactic acid bacteria. 100-1 Masmeen 107 Matjes. for chilled and frozen prepared fish products 47-9 for chilled and frozen raw fish 19-20 for crustacean shellfish 87-8 for cured. to reduce rate of fish spoilage 12 Ikan kaju. 151-3 detection methods 153 Liquid smoke 107-8 Listeria spp. causing spoilage in vacuum-packed dried salmon 111 Lactic acid fermentation. smoked and dried fish (Table) 116-7 for shellfish products 70 Microbiological limits. hot-smoking 107 preserved by dry/wet salting 99 Marinating 94. as hazard in regard to reef-caught fish 28 growth and survival in crustacean shellfish 84 pathogen profile 229-30 Listeria monocytogenes. leading to oxidative rancidity 11 Lipophilic shellfish poisoning toxins 149. for fresh and frozen fish (Table) 20 Micrococcus spp. enabled by carbohydrate source in fish products 130. definition 1 Legislation. growth and survival in chilled and frozen prepared fish products 46 growth and survival in cured. as autochthonous bacterial flora of fish 3 Lean fish.
contamination leading to illness 68 description and uses 57 effects of processing on microflora 60-4 hot-smoking 107 spoilage organisms 65 P. caused by lipid oxidation 11 Oysters.FISH AND SEAFOOD of crustacean shellfish 80-1 of cured. of crustacean shellfish – effects of processing 81-3 of cured. in crustacean shellfish 867 Norwalk virus. 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. pathogenic microorganisms in molluscan shellfish 59-60 Opisthorchis viverrini. shigelloides. 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. contaminated by azaspiracids 158 description and uses 58 effects of processing on microflora 60-4 hot-smoking 107 Nam-budu. as TTX producer 145 Norovirus. for detection of azaspiracids 160 for detection of ciguatoxins 157 for detection of DA 148 for detection of DSP toxins 150. associated with consumption of fermented fish products 137 Oxidative rancidity. fish sauce 123 Neosaxitoxin (neoSTS) 142-3 Neurotoxin shellfish poisoning (NSP) toxins 153-5 detection methods 155 Nitrosamines. reduction in smoking process 108 Norcardiopsis dassonvillei. smoked and dried fish 95 of fermented fish 126-7 of molluscan shellfish 59-60 Microflora. as autochthonous bacterial flora of fish 3 as spoilage organism in temperate crustacean shellfish 80 in molluscan shellfish 60 Mouse bioassay. growth and survival in chilled and frozen raw fish 17 Paralytic shellfish poisoning (PSP) toxins 142-4 detection methods 144 toxicology 143 252 .
81 as spoilage organism in fresh fish 13 as spoilage organism in oysters 65 as TTX producers 145 253 . 69-70 Polycyclic aromatic hydrocarbons. spoilage organism in oysters 65 Protoceratium reticulatum. adverse condition on poorly preserved fish 110 Pinnatoxins member of cyclic imine group 164. 161. reduction in smoking process 103. mould causing spoilage of dried fish 110 Potato. as component for seafood ready meals 40 Pathogen profiles. 197 Plesiomonas. fermented fish product 124 Plankton. in fish fillet 1 Proteus. pathogen profile 230 in molluscan shellfish 5960. detection methods 165 origins and distribution 165 toxicology 165 Pissala. as component for seafood ready meals 40 Prawns. as cause of histamine fish poisoning 15 as spoilage organism in chilled products 43 as spoilage organism in fresh fish 14-5 Pickling 94. 152-3 Penicillium spp. 163. as autochthonous bacterial flora of fish 3 as spoilage organism in chilled products 43 as spoilage organism in crustacean shellfish 80. in molluscan shellfish 60 Pseudomonas spp. growth and survival in chilled and frozen raw fish 19 growth and survival in crustacean shellfish 86 growth and survival in cured. causing spoilage of dried fish 110-1 Photobacterium phosphoreum. 108 Polypaecilum pisce. planktonic source of azaspiracids 159 Pseudomonadaceae. effects on microflora of fish 5-9 Professional bodies. preserved by salt-boiling 100 Pink.INDEX Parasites. contact details 241-2 Prorocentrum spp. as source of toxins 159. as source of yessotoxins 152 Protoperidinium cassipes. smoked and dried fish 115 Pasta. 100-1 Pindang. causing spoilage in vacuum-packed dried fish 111 Pseudomonads. effects of processing on microflora 81-2 freshness indicators 83 Pre-column derivatisation. 145-6 definitions 28-30 non-seafood ingredients 32 Processing. as source of DSP toxins 148-9 Protein content. smoked and dried fish 113-6 growth and survival in fermented fish 136-7 growth and survival in molluscan shellfish 66-70 Pectenotoxins 151. 225-39 Pathogens. for detection of PSP toxins 144. 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.
) 41 ingredients preparation. smoked and dried fish 114 pathogen profile 230-1 pathogenic microorganisms in molluscan shellfish 59-60 Salt burn. processing 35 Shellstock. smoking and marinating of fish 96 Salt-boiling 94. processing 34. growth and survival in chilled and frozen raw fish 19 Pseudoterranova dicipiens. preserved in saturated brine 99 Sauce. vacuum packing 105 Salmonella spp. importance in lactic acid fermentation in fish products 130 importance in salting. as source of tetrotoxin 144-5 Putty fish. flow diagram (Fig. hazard in salting of fish 97 Salt concentration. source of PSP toxins 142 Quality Index Method. detection methods 165 member of cyclic imine group 164 origins and distribution 165 toxicology 165 Puffer fish. description and uses 58-9 Scampi. initial microflora 31 Seafood ready meals 39-40 Seafood recipe meal centres 38-9 Shaped fish products. processing and effects on microflora 60-1 Shewanella. for evaluation of fish quality 9 Radiation.FISH AND SEAFOOD inhibition of growth by carbon dioxide 6 Pseudonitzschia multiseries. assembly and packaging. in molluscan shellfish 60 Rice. fermented fish product 126 as source of botulism 136 Rakørret. for fish ready meals – manufacture 40-2 Saxitoxin (STX) 142-3 Scallops.) 41 Recipe fish products 38-40 Rhodotorula rubra. flow diagram (Fig. as autochthonous bacterial flora of fish 3 inhibition of growth by carbon dioxide 6 Shewanella putrefaciens. 83 254 . chilled and frozen 1-25 Recipe dish. 36 Seafood. resulting from poorly salted cod 110 Pyrodinium. as spoilage organism in chilled products 43 as spoilage organism in fresh fish 13-4 as spoilage organism in temperate crustacean shellfish 80. producers of brevetoxins 154 Raw fish. 135 Rakefisk. for microbial inactivation in fish products 127. fermented fish product 126 Raphidophytes. to measure fish skin resistance 10 Salmon. detected in cod 115 Pteriatoxins. antibody-based kit to detect STX 144 RT Freshmeter. implicated in amnesic shellfish poisoning 146-7 Pseudoterranova. 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. as carbohydrate source in lactic acid fermentation in fish products 134 as component for seafood ready meals 40 RIDASCREEN.
in poorly salted cod 110 Smoked fish 93-121 Smoking 103-8 different processes 94-5 Snow Mountain virus. smoked and dried fish 114 pathogenic microorganisms in molluscan shellfish 59-60 Shottsuru. 13. growth and survival in chilled and frozen prepared fish products 46 growth and survival in cured. as indicator of fresh fish spoilage 11 Toxins. of fish after capture 5 Surstrømming. processing and effects on microflora 63-4 Skin resistance. contact details 241-2 Trematode infections foodborne 137 Trichosporon. of dried fish 111 of fish 6-7 of fish – (US) NACMCF recommendations 7-8 of fish – Codex Committee 255 . of cooked fish 9-10 Tetrotoxin 144-6 detection methods 145-6 toxicology 145 TMAO. as measure of fish degradation 10 Sliming. as contaminant in scampi 36 as contaminant in enrobing process 37 as hazard in dry salting of fish 89 growth and survival in chilled and frozen prepared fish products 46-7 growth and survival in crustacean shellfish 84 growth and survival in cured. of crustacean shellfish 80.INDEX Shigella spp. role in fish degradation 10. concerning crustacean shellfish 89 Vacuum packaging. 83 of fermented fish 135-6 of fish subjected to different preservation methods 108-10 of fresh fish 9-11. grading of 31 hot-smoking 107 US legislation. 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. in molluscan shellfish 60 Tuna. definitions of forms for purchase 79-80 effects of processing on microflora 81-2 freshness indicators 83 Shucked shellfish. fermented fish product 126 Taste panel evaluation. 14 Torry Scale. fish sauce 123 Shrimps. detection methods 164 member of cyclic imine group 162-4 origins and distribution 163 toxicology 163-5 Spoilage. pathogenic microorganisms in molluscan shellfish 59-60 Specific spoilage organisms (SSOs) 12-5 Spirolides. to measure fish skin resistance 10 Total Volatile Base-Nitrogen. fish and shellfish 141-74 Trade associations. for evaluation of fish quality 9 Torrymeter. 101 Storage. 12-5 of recipe fish meals 43-4 of shellfish 65 Staphylococcus.
processing for seafood recipe meal centres 39 Vibrio spp. as autochthonous bacterial flora of fish 3 as spoilage organisms in chilled products 43 as spoilage organisms in vacuum-packed dried fish 111 Viruses. 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. 152-3 256 . growth and survival in chilled and frozen raw fish 17 Vibrio cholerae 01 and 0139.FISH AND SEAFOOD guidance 7 of fresh chilled products 42 Vegetables. as spoilage organisms in chilled recipe meals 43 Yessotoxins 151. growth and survival in chilled and frozen raw fish 16-7 growth and survival in crustacean shellfish 86 growth and survival in fermented fish 136 growth and survival in molluscan shellfish 66-67 pathogen profile 234-5 Vibrio vulnificus. 68 Vibrio cholerae non-01. 81 as TTX producers 145 pathogenic microorganisms in molluscan shellfish 59-60 risk in cold-smoking 105 Vibrio cholerae. growth and survival in molluscan shellfish 67-8 Vibrio parahaemolyticus. growth and survival in chilled and frozen prepared fish products 47 growth and survival in crustacean shellfish 86-7 Wallemia sebi. growth and survival in crustacean shellfish 85 pathogen profile 233-4 Vibrio cholerae 01. as hazard in regard to reef-caught fish 28 as spoilage organism in crustacean shellfish 80. 62 Yersinia spp. of shellstock 61.