CHAPTER 13: Clostridium botulinum Toxin Formation

This guidance represents the Food and Drug Administration’s (FDA’s) current thinking on this topic. It does not create or confer any rights for or on any person and does not operate to bind FDA or the public. You can use an alternative approach if the approach satisfies the requirements of the applicable statutes and regulations. If you want to discuss an alternative approach, contact the FDA staff responsible for implementing this guidance. If you cannot identify the appropriate FDA staff, call the telephone number listed on the title page of this guidance.

UNDERSTAND THE POTENTIAL HAZARD. • Controlling the amount of moisture that is available for pathogenic bacteria growth Clostridium botulinum (C. botulinum) toxin (water activity) in the product by formulation formation can result in consumer illness and (covered in this chapter); death. It is the toxin responsible for botulism. • Controlling the amount of moisture that is About 10 outbreaks of foodborne botulism occur available for pathogenic bacteria growth annually in the United States, from all sources. (water activity) in the product by drying Symptoms include: weakness, vertigo, double (covered in Chapter 14); vision, difficulty in speaking, swallowing and • Controlling the introduction of pathogenic breathing, abdominal swelling, constipation, bacteria after the pasteurization process paralysis, and death. Symptoms start from 18 and after the cooking process performed hours to 36 hours after consumption. Everyone immediately before reduced oxygen packaging is susceptible to intoxication by C. botulinum (covered in Chapter 18); toxin; only a few micrograms of the toxin can • Controlling the source of molluscan shellfish cause illness in a healthy adult. Mortality is high; and the time from exposure to air (e.g., by without the antitoxin and respiratory support, harvest or receding tide) to refrigeration death is likely. to control pathogens from the harvest area This chapter covers the hazard of C. botulinum (covered in Chapter 4); growth and toxin formation as a result of time and • Managing the amount of time that food is temperature abuse during processing, storage, and exposed to temperatures that are favorable distribution. for pathogenic bacteria growth and toxin production (covered generally in Chapter • Strategies for controlling pathogen growth 12; for C. botulinum, in this chapter; and for There are a number of strategies for the control Staphylococcus aureus (S. aureus) in hydrated of pathogens in fish and fishery products. They batter mixes, in Chapter 15); include: • Killing pathogenic bacteria by cooking or • Controlling the level of acidity (pH) in the pasteurization (covered in Chapter 16), or product (covered by the Acidified Foods retorting (covered by the Thermally Processed regulation, 21 CFR 114, for shelf-stable Low-Acid Foods Packaged in Hermetically acidified products, and by this chapter for Sealed Containers regulation, 21 CFR 113 refrigerated acidified products); (hereinafter, the Low-Acid Canned Foods • Controlling the amount of salt or (LACF) Regulation)); preservatives, such as sodium nitrite, in the • Killing pathogenic bacteria by processes that product (covered in this chapter); retain the raw product characteristics (covered in Chapter 17).

CHAPTER 13: Clostridium botulinum Toxin Formation 245 • Formation of C. botulinum toxin packaging and modified atmosphere packaging) When C. botulinum grows, it can produce a extend the shelf life of a product by inhibiting potent toxin, one of the most poisonous naturally the growth of aerobic spoilage bacteria. There occurring substances known. The toxin can be is a safety concern with these products because destroyed by heat (e.g., boiling for 10 minutes), there is an increased potential for the formation but, because of its potency, you should not rely of C. botulinum toxin before spoilage makes the on this as a means of control. product unacceptable to consumers. The strains of C. botulinum can be divided C. botulinum forms toxin more rapidly at higher into two groups, the proteolytic group (i.e., temperatures than at lower temperatures. The those that break down proteins) and the non­ minimum temperature for growth and toxin proteolytic group (i.e., those that do not break formation by C. botulinum type E and non­ down proteins). The proteolytic group includes proteolytic types B and F is 38°F (3.3°C). For C. botulinum type A and some of types B and F. type A and proteolytic types B and F, the The non-proteolytic group includes C. botulinum minimum temperature for growth is 50°F (10°C). type E and some of types B and F. As the shelf life of refrigerated foods is increased, more time is available for C. botulinum growth The vegetative cells of all types of C. botulinum and toxin formation. As storage temperatures are easily killed by heat. However, C. botulinum increase, the time required for toxin formation is is able to produce spores. In this state, the significantly shortened. You should expect that at pathogen is very resistant to heat. The spores some point during storage, distribution, display, of the proteolytic group are much more resistant or consumer handling of refrigerated foods, safe to heat than are those of the non-proteolytic refrigeration temperatures will not be maintained group (i.e., they require a canning process to be (especially for the non-proteolytic group). Surveys destroyed). Table A-4 (Appendix 4) provides of retail display cases indicate that temperatures guidance about the conditions under which of 45 to 50°F (7 to 10°C) are not uncommon. the spores of the most heat-resistant form Surveys of home refrigerators indicate that of non-proteolytic C. botulinum, type B, are temperatures can exceed 50°F (10°C). killed. However, there are some indications that substances that may be naturally present In reduced oxygen packaged products in which in some products (e.g., dungeness crabmeat), the spores of non-proteolytic C. botulinum such as lysozyme, may enable non-proteolytic are inhibited or destroyed (e.g., smoked fish, C. botulinum to more easily recover after heat pasteurized crabmeat, and pasteurized surimi), damage, resulting in the need for a considerably a normal refrigeration temperature of 40°F more stringent process to ensure destruction. (4.4°C) is appropriate because it will limit the growth of proteolytic C. botulinum and C. botulinum is able to produce toxin when other pathogens that may be present. Even a product in which it is present is exposed to in pasteurized products where non-proteolytic temperatures favorable for growth for sufficient C. botulinum is the target organism for the time. Table A-1 (Appendix 4) provides guidance pasteurization process, and vegetative pathogens, about the conditions under which C. botulinum such as Listeria monocytogenes, are not likely and other pathogenic bacteria are able to grow. to be present (e.g., pasteurized crabmeat and Table A-2 (Appendix 4) provides guidance about pasteurized surimi), a storage temperature of the time necessary at various temperatures for 40°F (4.4°C) is still appropriate because of the toxin formation to occur. potential for survival through the pasteurization Packaging conditions that reduce the amount process and recovery of spores of non-proteolytic of oxygen present in the package (e.g., vacuum C. botulinum, aided by naturally occurring

CHAPTER 13: Clostridium botulinum Toxin Formation 246 substances, such as lysozyme. In this case, toxin formation in reduced oxygen packaged fish refrigeration serves as a prudent second barrier. and fishery products. However, in reduced oxygen packaged products Alternatively, products of this type may be safely in which refrigeration is the sole barrier to marketed frozen, with appropriate labeling outgrowth of non-proteolytic C. botulinum and to ensure that it is held frozen throughout the spores have not been destroyed (e.g., vacuum- distribution. For some reduced oxygen packaged packaged refrigerated raw fish, vacuum-packaged products, control of C. botulinum can be refrigerated unpasteurized crayfish meat, and achieved by breaking the vacuum seal before the reduced oxygen packaged unpasteurized product leaves the processor’s control. dungeness crabmeat), the temperature should be The guidance in this chapter emphasizes maintained below 38°F (3.3°C) from packing to consumption. Ordinarily you, as a processor, can preventive measures for the control of non­ ensure that temperatures are maintained below proteolytic strains of C. botulinum in products 38°F (3.3°C) while the product is in your control. that are contained in reduced oxygen packaging. However, the current U.S. food distribution As was previously described, this emphasis system does not ensure the maintenance of these is because such an environment extends the temperatures after the product leaves your control. shelf life of a refrigerated product in a way that, under moderate temperature abuse, favors C. The use of a Time-Temperature Indicator (TTI) on botulinum growth and toxin formation over each consumer package may be an appropriate aerobic spoilage. It is also possible for both means of overcoming these problems in the non-proteolytic and proteolytic C. botulinum distribution system for reduced oxygen packaged to grow and produce toxin in a product that is products in which refrigeration is the sole barrier not reduced oxygen packaged and is subjected to outgrowth of non-proteolytic C. botulinum and to severe temperature abuse. This is the case in which the spores have not been destroyed. because of the development within the product A TTI is a device that monitors the time and of microenvironments that support its growth. temperature of exposure of the package and However, this type of severe temperature abuse alerts the consumer or end user if a safe exposure of refrigerated products is not reasonably likely limit has been exceeded. If a TTI is used, it to occur in the processing environment of most should be validated to ensure that it is fit for its fish or fishery products and the Current Good intended purpose and verified that it is functional Manufacturing Practice in Manufacturing, Packing, at the time of use. It should be designed to alert or Holding Human Food regulation, 21 CFR 110, the consumer (e.g., a color change) that an unsafe requires refrigeration of foods that support the time and temperature exposure has occurred growth of pathogenic microorganisms. that may result in C. botulinum toxin formation. Additionally, the alert should remain perpetually • Sources of C. botulinum visible after it has been triggered, regardless of C. botulinum can enter the process on raw environmental conditions that could reasonably materials. The spores of C. botulinum are very be expected to occur thereafter. Skinner, G. E., common. They have been found in the gills and J. W. Larkin in “Conservative prediction of and viscera of finfish, crabs, and shellfish. C. time to Clostridium botulinum toxin formation for botulinum type E is the most common form found use with time-temperature indicators to ensure in freshwater and marine environments. Types the safety of foods,” Journal of Food Protection, A and B are generally found on land but may 61:1154-1160 (1998), describe a safe time and temperature exposure curve (“Skinner-Larkin also be occasionally found in water. It should be curve”) that may be useful in evaluating the assumed that C. botulinum will be present in any suitability of a TTI for control of C. botulinum raw fishery product, particularly in the viscera.

CHAPTER 13: Clostridium botulinum Toxin Formation 247 Because spores are known to be present in the packaging) may be rapidly depleted by the viscera, any product that will be preserved by activity of spoilage bacteria, resulting in the salting, drying, pickling, or fermentation should formation of a reduced oxygen environment. be eviscerated prior to processing (see the Packaging that provides an oxygen transmission “Compliance Policy Guide,” Sec. 540.650). Without rate (in the final package) of at least 10,000 cc/ evisceration, toxin formation is possible during the m2/24 hours at 24ºC can be regarded as an process, even with strict control of temperature. oxygen-permeable packaging material for fishery Evisceration of fish is the careful and complete products. The oxygen transmission rate of removal of all internal organs in the body cavity packaging material is listed in the packaging without puncturing or cutting them, including specifications that can be obtained from the gonads. If even a portion of the viscera or its packaging manufacturer. contents is left behind, the risk of toxin formation by C. botulinum remains. Uneviscerated small An oxygen-permeable package should provide fish, less than 5 inches in length (e.g., anchovies sufficient exchange of oxygen to allow aerobic and herring sprats), for which processing eliminates spoilage organisms to grow and spoil the product preformed toxin, prevents toxin formation during before toxin is produced under moderate abuse processing and that reach a water phase salt temperatures. Particular care should be taken in content of 10% in refrigerated finished products, determining the safety of a packaging material for a or a water activity of below 0.85 in shelf-stable product in which the spoilage organisms have been finished products, or a pH of 4.6 or less in shelf- eliminated or significantly reduced by processes stable finished products, are not subject to the such as high pressure processing. The generally evisceration recommendation. recommended 10,000 cc/m2/24 hours at 24ºC transmission rate may not be suitable in this case. Note: The water phase salt content of 10% is based on the control of C. botulinum type A and proteolytic types B and F. Use of an oxygen-permeable package may not Note: The water activity value of below 0.85 is based on the compensate for the restriction to oxygen exchange minimum water activity for toxin production of S. aureus. created by practices such as packing in oil or in • Reduced oxygen packaging deep containers from which the air is expressed or the use of oxygen scavengers in the packaging. A number of conditions can result in the creation of a reduced oxygen environment in packaged • Control of C. botulinum fish and fishery products. They include: There are a number of strategies to prevent C. • Vacuum, modified, or controlled atmosphere botulinum growth and toxin formation during packaging. These packaging methods processing, storage, and distribution of finished generally directly reduce the amount of fish and fishery products. They include: oxygen in the package; For products that do not require refrigeration • Packaging in hermetically sealed containers (i.e., shelf-stable products): (e.g., double-seamed cans, glass jars • Heating the finished product in its final with sealed lids, and heat-sealed plastic container sufficiently by retorting to destroy containers), or packing in deep containers the spores of C. botulinum types A B, E, from which the air is expressed (e.g., caviar and F (e.g., canned fish). This strategy is in large containers), or packing in oil. These covered by the LACF Regulation, 21 CFR and similar processing and packaging 113, and these controls are not required to techniques prevent the entry of oxygen into be included in your Hazard Analysis Critical the container. Any oxygen present at the Control Point (HACCP) plan; time of packaging (including oxygen that may be added during modified atmosphere

CHAPTER 13: Clostridium botulinum Toxin Formation 248 • Controlling the level of acidity (pH) in the • Controlling the amount of salt in the product finished product to 4.6 or below, to prevent to 20% water phase salt (wps) or more, to growth and toxin formation by C. botulinum prevent the growth of C. botulinum types A, types A, B, E, and F (e.g., shelf-stable B, E, and F and other pathogens that may acidified products). This strategy is covered be present in the product (e.g., shelf-stable by the Acidified Foods regulation, 21 CFR salted products). This strategy is covered 114, and these controls are not required to be in this chapter. Water phase salt is the included in your HACCP plan; concentration of salt in the water-portion of • Controlling the amount of moisture that is the fish flesh and calculated as follows: (% available in the product (water activity) to NaCl X 100)/(% NaCl + % moisture) = % NaCl 0.85 or below by drying, to prevent growth in water phase. The relationship between and toxin formation by C. botulinum types percent water phase salt and water activity in A, B, E, and F and other pathogens that may fish is described in the following graph. be present in the product (e.g., shelf-stable dried products). This strategy is covered by Chapter 14;

Relationship of Water Activity to Water Phase 1 Salt in NaCl/Water Solutions

1.00 0.98

y 0.96 it

tivc 0.94

r a 0.92 tea 0.90 W 0.88 0.86 0.84 0 2 4 6 8 10 12 14 16 18 20 Percent water phase salt

1. This relationship is generally valid for fish products when salt (sodium chloride) is the primary means of binding water. The specific food matrix and the use of other salts or water binding agents could affect the exact relationship. If you intend to use this relationship in your control strategy, you should determine the exact relationship in your product by conducting a study.

CHAPTER 13: Clostridium botulinum Toxin Formation 249 For products that require refrigeration: type A and proteolytic types B and F and • Heating the finished product in its final other pathogens that may be present in container sufficiently by pasteurization to the finished product through refrigerated destroy the spores of C. botulinum type storage (e.g., refrigerated dried fish). Drying E and non-proteolytic types B and F, and is covered in Chapter 14, controlling the then minimizing the risk of recontamination growth of proteolytic C. botulinum through by controlling seam closures and cooling refrigeration is covered in this chapter, and water, and next controlling the growth controlling the growth of other pathogenic of the surviving C. botulinum type A and bacteria through refrigeration is covered in proteolytic types B and F in the finished Chapter 12; product with refrigerated storage (e.g.. • Controlling the level of pH to 5 or below, salt pasteurized crabmeat and some pasteurized to 5% wps or more, moisture (water activity) surimi-based products). Pasteurization to 0.97 or below, or some combination is covered in Chapter 16, controlling of these barriers, in the finished product recontamination after pasteurization is sufficiently to prevent the growth of C. covered in Chapter 18, and controlling the botulinum type E and non-proteolytic growth of proteolytic C. botulinum through types B and F by formulation, and then refrigeration is covered in this chapter; controlling the growth of C. botulinum • Heating the product sufficiently to destroy type A and proteolytic types B and F and the spores of C. botulinum type E and other pathogens that may be present in non-proteolytic types B and F, and then the finished product with refrigerated minimizing the risk of recontamination storage (e.g., refrigerated acidified (pickled) by hot filling the product into the final products). Controlling the growth of non- container in a sanitary, continuous, closed proteolytic C. botulinum through formulation filling system and controlling seam closures is covered in this chapter, controlling the and cooling water, and next controlling growth of proteolytic C. botulinum through the growth of the surviving C. botulinum refrigeration is covered in this chapter, and type A and proteolytic types B and F and controlling the growth of other pathogenic other pathogens that may be present in the bacteria through refrigeration is covered in finished product with refrigerated storage Chapter 12; (e.g., vacuum packed soups, chowders, and • Controlling the amount of salt and sauces). Specialized cooking processes preservatives, such as sodium nitrite, in are covered in Chapter 16, prevention of the finished product, in combination with recontamination after specialized cooking other barriers, such as smoke, heat damage, processes is covered in Chapter 18, and competitive bacteria, sufficiently to controlling the growth of proteolytic C. prevent the growth of C. botulinum type botulinum through refrigeration is covered E and non-proteolytic types B and F, and in this chapter, and controlling the growth then controlling the growth of C. botulinum of other pathogenic bacteria through type A and proteolytic types B and F and refrigeration is covered in Chapter 12; other pathogens that may be present in the • Controlling the amount of moisture that finished product with refrigerated storage is available in the product (water activity) (e.g., salted, smoked, or smoke-flavored fish). to 0.97 or below to inhibit the growth of Controlling the growth of non-proteolytic C. botulinum type E and non-proteolytic C. botulinum through salting and smoking types B and F by drying, and then is covered in this chapter, controlling the controlling the growth of C. botulinum growth of proteolytic C. botulinum through

CHAPTER 13: Clostridium botulinum Toxin Formation 250 refrigeration is covered in this chapter, and In hot-smoked products, heat damage to controlling the growth of other pathogenic the spores of C. botulinum type E and non­ bacteria through refrigeration is covered in proteolytic types B and F also helps prevent Chapter 12; toxin formation. In these products, control of • Controlling the amount of salt in the the heating process is critical to the safety of finished product, in combination with heat the finished product. It is important to note, damage from pasteurization in the finished however, that this same heating process also product container, sufficiently to prevent reduces the numbers of naturally occurring the growth of C. botulinum type E and spoilage organisms. The spoilage organisms nonproteolytic types B and F, and then would otherwise have competed with, and controlling the growth of C. botulinum inhibited the growth of, C. botulinum. type A and proteolytic types B and F and In cold-smoked fish, it is important that other pathogens that may be present in the product does not receive so much heat the finished product with refrigerated that the numbers of spoilage organisms storage (e.g., some pasteurized surimi­ are significantly reduced. This is important based products). Controlling the growth because spoilage organisms must be present of non-proteolytic C. botulinum through to inhibit the growth and toxin formation a combination of salt and heat damage of C. botulinum type E and non-proteolytic is covered in this chapter, controlling the types B and F. This inhibition is important growth of proteolytic C. botulinum through in cold-smoked fish because the heat applied refrigeration is covered in this chapter, and during this process is not adequate to controlling the growth of other pathogenic weaken the C. botulinum spores. Control bacteria through refrigeration is covered in of the temperature during the cold-smoking Chapter 12. process to ensure survival of the spoilage Examples of C. botulinum control in specific organisms is, therefore, critical to the safety products: of the finished product. The interplay of these inhibitory effects • Refrigerated (not frozen), reduced oxygen (i.e., salt, temperature, smoke, and nitrite) packaged smoked and smoke-flavored fish is complex. Control of the brining or dry Achieving the proper concentration of salting process is clearly critical to ensure that salt and nitrite in the flesh of refrigerated, there is sufficient salt in the finished product. reduced oxygen packaged smoked and However, preventing toxin formation by C. smoke-flavored fish is necessary to prevent botulinum type E and non-proteolytic types the formation of toxin by C. botulinum type B and F is made even more complex by the E and non-proteolytic types B and F during fact that adequate salt levels are not usually storage and distribution. Salt works along achieved during brining. Proper drying with smoke and any nitrites that are added during smoking is also critical in order to to prevent growth and toxin formation by C. achieve the finished product water phase botulinum type E and non-proteolytic types salt level (i.e., the concentration of salt in B and F. Note that nitrites should be used the water portion of the fish flesh) needed only in salmon, sable, shad, chubs, and tuna, to inhibit growth and toxin formation by C. according to 21 CFR 172.175 and 21 CFR botulinum. 172.177 , and should not exceed a level of 200 ppm in salmon, sable, shad, chubs and This chapter covers the control procedures 10 ppm in tuna. described above.

CHAPTER 13: Clostridium botulinum Toxin Formation 251 You should ordinarily restrict brining, dry also serves as a prudent second barrier salting, and smoking loads to single species because of the potential survival through and to fish portions of approximately the pasteurization process and recovery of uniform size. This restriction minimizes the spores of non-proteolytic C. botulinum, aided complexity of controlling the operation. You by naturally occurring substances, such as should treat brine to minimize microbial lysozyme. Cooking and pasteurization are contamination or periodically replace it as a covered in Chapter 16, and controlling the good manufacturing practice control. growth of C. botulinum through refrigeration is covered in this chapter. The combination of inhibitory effects that are present in smoked and smoke-flavored fish In the second category of products, filling are not adequate to prevent toxin formation the product into the final container while it is by C. botulinum type A and proteolytic types still hot in a continuous, closed filling system B and F. Strict refrigeration control (i.e., at (i.e., hot filling) is also critical to the safety of or below 40°F (4.4°C)) during storage and the finished product because it minimizes the distribution should be maintained to prevent risk of recontamination of the product with growth and toxin formation by C. botulinum pathogens, including C. botulinum type E and type A and proteolytic types B and F and non-proteolytic types B and F. This control other pathogens that may be present in strategy applies to products such as soups, these products. Controlling the growth of chowders, and sauces that are filled directly proteolytic C. botulinum through refrigeration from the cooking kettle, where the risk of is covered in this chapter, and controlling the recontamination is minimized. It may not growth of other pathogenic bacteria through apply to products such as crabmeat, lobster refrigeration is covered in Chapter 12. meat, or crayfish meat or to other products that are handled between cooking and filling. • Refrigerated (not frozen), reduced oxygen Control of hot filling is covered in Chapter 18. packaged, pasteurized fishery products Chapter 18 also covers other controls that Refrigerated, reduced oxygen packaged, may be necessary to prevent recontamination, pasteurized fishery products fall into two including controlling container sealing and categories: (1) those which are pasteurized controlling contamination of container in the final container; and (2) those which cooling water. These controls may be critical are cooked in a kettle and then hot filled to the safety of both categories of products. into the final container in a continuous, closed filling system (e.g., heat-and-fill Examples of properly pasteurized products soups, chowders, and sauces). In both follow: fish and fishery products generally cases, ordinarily the heating process should (e.g., surimi-based products, soups, be sufficient to destroy the spores of C. or sauces) pasteurized to a minimum

botulinum type E and non-proteolytic types cumulative total lethality of F194°F (F90°C) B and F. In neither case is it likely that = 10 minutes, where z = 12.6°F (7°C) for the heating process will be sufficient to temperatures less than 194°F (90°C), and destroy the spores of C. botulinum type A z = 18°F (10°C) for temperatures above and proteolytic types B and F. Therefore, 194°F (90°C); blue crabmeat pasteurized strict refrigeration control (i.e., at or below to a minimum cumulative total lethality of

40°F (4.4°C)) should be maintained during F185°F (F85°C) = 31 minutes, where z = 16°F storage and distribution to prevent growth (9°C); and dungeness crabmeat pasteurized and toxin formation by C. botulinum type A to a minimum cumulative total lethality of

and proteolytic types B and F. Refrigeration F194°F (F90°C) = 57 minutes, where z = 15.5°F

CHAPTER 13: Clostridium botulinum Toxin Formation 252 (8.6°C). Equivalent processes at different • Adding sufficient acid to reduce temperatures can be calculated using the z the acidity (pH) to 5.0 or below; values provided. • Reducing the amount of moisture that is available for growth (water EXAMPLES OF PROPERLY PASTEURIZED activity) to below 0.97 (e.g., by PRODUCTS adding salt or other substances that MINIMUM CUMULATIVE PRODUCT Z VALUE “bind” the available water); or TOTAL LETHALITY • Making a combination of salt, pH, Fish and fishery F194°F (F90°C) = 10 minutes 12.6°F (7°C), for products temperatures and/or water activity adjustments generally less than 194°F (e.g., surimi­ (90°C) that, when combined, prevents the based products, 18°F (10°C) for growth of C. botulinum type E and soups, or temperatures sauces) above 194°F non-proteolytic types B and F (to be (90°C) established by a scientific study). Blue crabmeat F (F ) = 31 minutes 16°F (9°C) 185°F 85°C Much like smoked products, in some of these Dungeness F (F ) = 57 minutes 15.5°F (8.6°C) 194°F 90°C products the interplay of these inhibitory crabmeat effects (i.e., salt, water activity, and pH) can be complex. Control of the brining, pickling, In some pasteurized surimi-based or formulation steps is, therefore, critical to products, salt, in combination with a milder ensure that there are sufficient barriers in the pasteurization process, in the finished product finished product to prevent the growth and container works to prevent growth and toxin toxin formation of C. botulinum type E and formation by C. botulinum type E and non­ non-proteolytic types B and F during storage proteolytic types B and F. An example of a and distribution. These control procedures properly pasteurized surimi-based product are covered in this chapter. in which 2.4% wps is present is one that has been pasteurized at an internal temperature You should ordinarily restrict brining and of 185°F (85°C) for at least 15 minutes. This pickling loads to single species and to fish process may not be suitable for other types of portions of approximately uniform size. products because of the unique formulation This restriction minimizes the complexity of and processing involved in the manufacture of controlling the operation. You should treat surimi-based products. brine to minimize microbial contamination or periodically replace it as a good • Refrigerated (not frozen), reduced oxygen manufacturing practice control. packaged pickled fish, salted fish, caviar, and similar products The controls discussed above are not sufficient to prevent toxin formation by C. In pickled fish, salted fish, caviar, and similar botulinum type A and proteolytic types B products that have not been preserved and F. Strict refrigeration control (i.e., at sufficiently for them to be shelf stable, or below 40°F (4.4°C)) during storage and growth and toxin formation by C. botulinum distribution should, therefore, be maintained type E and non-proteolytic types B and F is to prevent growth and toxin formation by C. controlled by one of the following: botulinum type A and proteolytic types B and • Adding sufficient salt to produce F and other pathogens that may be present a water phase salt level (i.e., the in these products. Controlling the growth of concentration of salt in the water proteolytic C. botulinum through refrigeration portion of the fish flesh) of at least 5%; is covered in this chapter, and controlling the

CHAPTER 13: Clostridium botulinum Toxin Formation 253 growth of other pathogenic bacteria through • Frozen, reduced oxygen packaged raw, refrigeration is covered in Chapter 12. unpreserved fish and unpasteurized, cooked fishery products • Refrigerated (not frozen), reduced oxygen For frozen, reduced oxygen packaged raw, packaged raw, unpreserved fish and unpreserved fish (e.g., frozen, vacuum- unpasteurized, cooked fishery products packaged fish fillets) and frozen, reduced For refrigerated, reduced oxygen packaged oxygen packaged, unpasteurized, cooked raw, unpreserved fish (e.g., refrigerated, fishery products (e.g., frozen, vacuum- vacuum-packaged fish fillets) and refrigerated, packaged, unpasteurized crabmeat, lobster reduced oxygen packaged, unpasteurized, meat, or crayfish meat), the sole barrier to cooked fishery products (e.g., refrigerated, toxin formation by C. botulinum type E vacuum-packaged, unpasteurized crabmeat, and non-proteolytic types B and F during lobster meat, or crayfish meat), the sole finished product storage and distribution barrier to toxin formation by C. botulinum is freezing. Because these products may type E and non-proteolytic types B and appear to the retailer, consumer, or end user F during finished product storage and to be intended to be refrigerated, rather than distribution is refrigeration. These types of C. frozen, labeling to ensure that they are held botulinum will grow at temperatures as low frozen throughout distribution is critical to as 38°F (3.3°C). As was previously noted, their safety. maintenance of temperatures below 38°F (3.3°C) after the product leaves your control Controls should be in place to ensure that and enters the distribution system cannot such products are immediately frozen after normally be ensured. The use of a TTI on processing, maintained frozen throughout the smallest unit of packaging (i.e., the unit storage in your facility, and labeled to of packaging that will not be distributed be held frozen and to be thawed under any further, usually consumer or end-user refrigeration immediately before use (e.g., package) may be an appropriate means of “Important, keep frozen until used, thaw overcoming these problems in the distribution under refrigeration immediately before use”). system. This chapter provides controls for the Frozen, reduced oxygen packaged products application of TTIs for packaging. that are customarily cooked by the consumer or end user in the frozen state (e.g., boil-in­ If you intend to package these products in bag products and frozen fish sticks) need not a reduced oxygen package and you do not be labeled to be thawed under refrigeration. intend to apply a TTI on each consumer For purposes of hazard analysis, other frozen package, you should evaluate the effectiveness products that do not contain the “keep of other preventive measures, either singularly, frozen” statement should be evaluated as if or in combination, that may be effective in they will be stored refrigerated because the preventing growth and toxin formation by C. consumer or end user would not have been botulinum. Such evaluation is customarily warned to keep them frozen. accomplished by conducting an inoculated pack study under moderate abuse conditions. Control procedures to ensure that product A suitable protocol for the performance of is properly labeled with “keep frozen” such studies is contained in a 1992 publication instructions are covered in this chapter. by the National Advisory Committee on Microbiological Criteria for Foods, “Vacuum or modified atmosphere packaging for refrigerated, raw fishery products.”

CHAPTER 13: Clostridium botulinum Toxin Formation 254 • Control in unrefrigerated (shelf-stable), DETERMINE WHETHER THE POTENTIAL reduced oxygen packaged fishery products HAZARD IS SIGNIFICANT. Examples of shelf-stable, reduced oxygen packaged fishery products are dried fish, The following guidance will assist you in acidified fish, canned fish, and salted fish. determining whether C. botulinum toxin formation Because these products are marketed without is a significant hazard at a processing step: refrigeration, either (1) the spores of C. botulinum types A, B, E, and F should be 1. Is it reasonably likely that C. botulinum will destroyed after the product is placed in the grow and produce toxin during finished product finished product container (covered by the storage and distribution? LACF Regulation, 21 CFR 113) or (2) a barrier, The factors that make C. botulinum toxin or combination of barriers, should be in place formation during finished product storage that will prevent growth and toxin formation and distribution reasonably likely to occur by C. botulinum types A, B, E, and F, and are those that may result in the formation of other pathogens that may be present in the a reduced oxygen packaging environment. product. Suitable barriers include: These are discussed in the section • Adding sufficient salt to produce “Understand the potential hazard,” under the a water phase salt level (i.e., the heading, “Reduced oxygen packaging.” concentration of salt in the water 2. Can growth and toxin formation by C. botulinum that portion of the fish flesh) of at least is reasonably likely to occur be eliminated or reduced 20%. Note that this value is based on to an acceptable level at this processing step? the maximum salt level for growth of S. aureus, covered in this chapter; C. botulinum toxin formation should also be considered a significant hazard at any • Reducing the amount of moisture processing step where a preventive measure that is available for growth (water is, or can be, used to eliminate the hazard activity) to below 0.85 (e.g., by adding (or reduce the likelihood of its occurrence to salt or other substances that bind the an acceptable level) if it is reasonably likely available water). Note that this value to occur. is based on the minimum water activity for growth and toxin formation of S. Preventive measures for C. botulinum toxin aureus, covered in this chapter; formation during finished product distribution and storage are discussed in the section, • Adding sufficient acid to reduce the pH “Understand the potential hazard,” under the to 4.6 or below. This barrier is covered heading, “Control of C. botulinum.” by the Acidified Foods regulation, 21 CFR 114, and these controls are not required • Intended use to be included in your HACCP plan; Because of the extremely toxic nature of • Drying the product sufficiently to C. botulinum toxin, it is unlikely that the reduce the water activity to 0.85 or significance of the hazard will be affected by the below. Note that this value is based intended use of your product. on the minimum water activity for growth and toxin formation of S. aureus, covered in Chapter 14.

Note: A heat treatment, addition of chemical additives, or other treatment may be necessary to inhibit or eliminate spoilage organisms (e.g., mold) in shelf-stable products.

CHAPTER 13: Clostridium botulinum Toxin Formation 255 IDENTIFY CRITICAL CONTROL POINTS. • Products dried to control the growth of C. botulinum type E The following guidance will assist you in and non-proteolytic types B and determining whether a processing step is a F and refrigerated to control the critical control point (CCP) for C. botulinum toxin growth of C. botulinum type A and formation: proteolytic types B and F and other pathogens that may be present. 1. Is there an acidification step (equilibrium pH In these cases, you should also identify of 4.6 or below), a drying step, an in-package the finished product storage step as pasteurization step, a combination of cook and a CCP for the hazard. Control of hot-fill steps, or a retorting step (commercial refrigeration is covered in this chapter for sterility) in the process? C. botulinum and in Chapter 12 for other a. If there is, you should in most cases pathogenic bacteria. identify the acidification step, drying Additionally, some pasteurized surimi­ step, pasteurization step, cook and hot- based products rely on a combination of fill steps, or retorting step as the CCP(s) salt and a relatively mild pasteurization for this hazard. Other processing steps process in the finished product container where you have identified C. botulinum for the control of C. botulinum type E toxin formation as a significant hazard and non-proteolytic types B and F. In will then not require control and will these products, you should also identify not need to be identified as CCPs for the formulation step as a CCP for the the hazard. However, control should hazard. Guidance provided in “Control be provided for time and temperature Strategy Example 4 - Pickling and Salting” exposure during finished product may be useful in developing controls at storage and distribution of the following this step. products: Guidance for the C. botulinum control • Products pasteurized in the final strategies listed above is contained in the container to kill C. botulinum type following locations: E and non-proteolytic types B • Control of cooking and hot-filling and F and refrigerated to control is covered in Chapters 16 and 18; the growth of C. botulinum type A and proteolytic types B and F • Control of pasteurization is and other pathogens that may covered in Chapters 16 and 18; be present (e.g., pasteurized • Control of drying is covered crabmeat and pasteurized surimi); in Chapter 14; • Products cooked to kill C. botulinum • Control of acidification is type E and non-proteolytic types covered in the Acidified Foods B and F, and then hot filled into regulation, 21 CFR 114; the final container, and next refrigerated to control the growth • Control of retorting is covered in of C. botulinum type A and the LACF Regulation, 21 CFR 113. proteolytic types B and F and other Note: Acidification and retorting controls for C. botulinum pathogens that may be present; required by 21 CFRs 113 and 114 need not be included in your HACCP plan.

CHAPTER 13: Clostridium botulinum Toxin Formation 256 b. If there is no acidification step and the smoking step should be identified as (equilibrium pH of 4.6 or below), drying a CCP for this hazard. The smoking step for step, pasteurization step, cooking and hot-smoked fish should be sufficient to damage hot-filling, or retorting (commercial the spores and make them more susceptible to sterility) step in the process, then decide inhibition by salt. The smoking step for cold- which of the following categories best smoked fish should not be so severe that it kills describes your product and refer to the the natural spoilage bacteria. These bacteria guidance below: are necessary so that the product will spoil before toxin production occurs. It is likely • Smoked and smoke-flavored fish; that they will also produce acid, which will • Fishery products in which further inhibit C. botulinum growth and toxin refrigeration is the sole barrier formation. to prevent toxin formation; This control approach is a control strategy • Fishery products in which freezing is referred to in this chapter as “Control Strategy the sole barrier to toxin formation; Example 1 - Smoking (1b - Cold Smoking and 1c - Hot Smoking).” • Pickled fish and similar products. 3. Is the storage temperature important to the safety • Smoked and smoke-flavored fish of the product? 1. Is the water phase salt level and, when permitted, the nitrite level, important to the safety of the Refrigerated (not frozen) finished product product? storage is critical to the safety of all products in this category and should be identified as For all products in this category, the water a CCP. Toxin formation by C. botulinum phase salt level is critical to the safety of the type A and proteolytic types B and F is not product, and the brining, dry salting and, inhibited by water phase salt levels below where applicable, drying steps should be 10%, nor by the combination of inhibitors identified as CCPs. Nitrite, when permitted, present in most smoked or smoke-flavored allows a lower level of salt to be used. Salt fish. Bacillus cereus can grow and form and nitrite are the principal inhibitors to toxin at water phase salt concentrations as C. botulinum type E and non-proteolytic high as 18%. types B and F toxin formation in these This control approach is a control strategy products. The water phase salt level needed referred to in this chapter as “Control Strategy to inhibit toxin formation is partially achieved Example 1 - Smoking (1d - Refrigerated during brining or dry salting and is partially Finished Product Storage).” achieved during drying. Control should be exercised over both operations. In some cases, salted, smoked, or smoke- flavored fish are received as ingredients This control approach is a control strategy for assembly into another product, such referred to in this chapter as “Control Strategy as a salmon paté. In other cases, they are Example 1 - Smoking (1a - Brining, Dry received simply for storage and further Salting, and Drying).” distribution (e.g., by a warehouse). In either 2. Is the temperature of the heating or smoking case, the refrigerated (not frozen) storage step process important to the safety of the product? is critical to the safety of the product and should be identified as a CCP. Control is the For both cold-smoked and hot-smoked fish same as that provided under “Control Strategy products, the temperature of smoking is critical, Example 1 - Smoking (1d - Refrigerated

CHAPTER 13: Clostridium botulinum Toxin Formation 257 Finished Product Storage).” Additionally, This control approach is a control strategy receiving of these products should be referred to in this chapter as “Control Strategy identified as a CCP, where control can be Example 2 - Refrigeration With a TTI (2e ­ exercised over the time and temperature Receipt of Product by Secondary Processor).” during transit. As previously noted, maintenance of This control approach is a control strategy temperatures below 38°F (3.3°C) after the referred to in this chapter as “Control product leaves your control and enters the Strategy Example 1 - Smoking (1e - Receipt of distribution system cannot normally be Products by Secondary Processor).” ensured. The use of a TTI on the smallest unit of packaging (i.e., the unit of packaging • Fishery products in which refrigeration is that will not be distributed any further, the sole barrier to prevent toxin formation usually consumer or end-user package) may 1. Is the storage temperature important to the safety be an appropriate means of overcoming these of the product? problems in the distribution system. When TTIs are used in this manner, their receipt, Refrigerated finished product storage is storage, and application and activation should critical to the safety of all products in this be identified as CCPs. category and should be identified as a CCP. These products contain no barriers (other This control approach is a control strategy than refrigeration) to toxin formation by C. referred to as “Control Strategy Example 2 ­ botulinum type E and non-proteolytic types Refrigeration With TTI (2a - Unactivated TTI B and F during finished product storage and Receipt, 2b - Unactivated TTI Storage, and 2c distribution. These types of C. botulinum - Application and Activation of TTI).” will grow at temperatures as low as 38°F • Fishery products in which freezing is the (3.3°C), necessitating particularly stringent sole barrier to toxin formation temperature control. 1. Is the storage temperature important to the safety This control approach is a control strategy of the product? referred to in this chapter as “Control Strategy Example 2 - Refrigeration With TTI (2d ­ Frozen finished product storage is critical to Refrigerated Finished Product Storage).” the safety of all products in this category. These products contain no barriers (other In some cases, these products are received as than freezing) to toxin formation by C. ingredients for assembly into another product. botulinum type E and non-proteolytic types In other cases, they are received simply for B and F during finished product storage and storage and further distribution (e.g., by a distribution. As previously noted, because warehouse). In either case, the refrigerated these products may appear to the retailer, storage step is critical to the safety of the consumer, or end user to be intended to be product and should be identified as a CCP. refrigerated, rather than frozen, labeling to Control is the same as that provided under ensure that they are held frozen throughout “Control Strategy Example 2 - Refrigeration distribution is critical to their safety and With a TTI (2d - Refrigerated Finished should be identified as a CCP. Product Storage).” Additionally, receiving of these products should be identified as a CCP, This control approach is a control strategy where control can be exercised over the time referred to in this chapter as “Control Strategy and temperature during transit. Example 3 - Frozen With Labeling.”

CHAPTER 13: Clostridium botulinum Toxin Formation 258 • Pickled and salted fish and similar products Salting (4b - Refrigerated Finished Product 1. Is the water phase salt level, water activity, and/ Storage).” Additionally, receiving of these or pH level important to the safety of the product? products should be identified as a CCP, where control can be exercised over time and For all products in this category, the water temperature during transit. phase salt level, water activity, and/or pH This control approach is a control strategy level are critical to the safety of the product referred to in this chapter as “Control Strategy because they are the principal inhibitors to Example 4 - Pickling and Salting (4c - Receipt growth and toxin formation by C. botulinum of Product by Secondary Processor).” type E and non-proteolytic type B and F. The levels of these inhibitors needed to inhibit toxin formation are achieved during the DEVELOP A CONTROL STRATEGY. pickling, brining, or formulation step. Control should be exercised over the relevant step. The following guidance provides four control strategies for C. botulinum toxin formation. You This control approach is a control strategy may select a control strategy that is different from referred to in this chapter as “Control Strategy those which are suggested, provided it complies Example 4 - Pickling and Salting (4a - with the requirements of the applicable food Brining, Pickling, Salting, and Formulation).” safety laws and regulations. Control strategies 2. Is the storage temperature important to the safety contain several elements that may need to be of the product? used in combination to result in an effective control program. Unless pickling, brining, or formulation results The following are examples of control strategies in a water phase salt level of at least 20% included in this chapter: (note that this value is based on the maximum

salt concentration for growth of S. aureus), a MAY APPLY TO MAY APPLY TO pH of 4.6 or below, or a water activity of 0.85 CONTROL STRATEGY PRIMARY SECONDARY PROCESSOR PROCESSOR or below (note that this value is based on Smoking   the minimum water activity for growth of S. aureus), refrigerated finished product storage Refrigeration with TTI   is critical to ensure the safety of the product Frozen with labeling   and should be identified as a CCP. Pickling and salting   This control approach is a control strategy referred to in this chapter as “Control Strategy • CONTROL STRATEGY EXAMPLE 1 - SMOKING Example 4 - Pickling and Salting (4b ­ This control strategy should include the following Refrigerated Finished Product Storage).” elements, as appropriate:

In some cases, pickled fish or similar a. Brining, dry salting, and drying; products are received as ingredients for assembly into another product. In b. Cold smoking; other cases, they are received simply for c. Hot smoking; storage and further distribution (e.g., by a warehouse). In either case, the refrigerated d. Refrigerated finished product storage; storage step is critical to the safety of the product and should be identified as a CCP. e. Receipt of products by secondary Control is the same as that provided under processor. “Control Strategy Example 4 - Pickling and

CHAPTER 13: Clostridium botulinum Toxin Formation 259 1A. BRINING, DRY SALTING, AND DRYING • Monitor brine temperature at the start of the brining process Set Critical Limits. with a temperature- indicating • The minimum or maximum values for the device (e.g., a thermometer), critical factors of the brining, dry salting, and then monitor ambient air and/or drying processes established by a temperature using a continuous scientific study. The critical factors are those temperature-recording device that are necessary to ensure that the finished (e.g., a recording thermometer); product has not less than 3.5% wps or, where AND permitted, the combination of 3% wps and not less than 100 ppm nitrite. The critical ° Monitor the drying time and the input/ factors may include: brine strength; brine to output air temperature (as specified fish ratio; brining time; brining temperature; by the study) using a continuous thickness, texture, fat content, quality, and temperature-recording device (e.g., a species of fish; drying time; input/output air recording thermometer); temperature, humidity, and velocity; smoke AND density; and drier loading. ° Monitor all other critical factors specified by the study with equipment appropriate Establish Monitoring Procedures. for the measurement; » What Will Be Monitored? OR • The critical factors of the established brining, • Collect a representative sample of the dry salting, and/or drying processes. These finished product and conduct water phase may include: brine strength; brine to fish salt analysis and, when appropriate, nitrite ratio; brining time; brining temperature; analysis. thickness, texture, fat content, quality, and species of fish; drying time; input/output air » How Often Will Monitoring Be Done (Frequency)? temperature, humidity, and velocity; smoke • For brine strength: density; and drier loading; ° At least at the start of the brining OR process; • The water phase salt and, where appropriate, AND nitrite level of the finished product. • For brine time: » How Will Monitoring Be Done? ° Once per batch; • For monitoring critical factors: AND ° Monitor brine strength with a • For manual brine temperature monitoring: salinometer; ° At the start of the brining process and at AND least every 2 hours thereafter; ° Monitor brine time with a clock; AND AND • For continuous temperature-recording devices: ° Monitor brine temperature using: ° Continuous monitoring by the device • A temperature-indicating device itself, with a visual check of the recorded (e.g., a thermometer); data at least once per batch; OR

CHAPTER 13: Clostridium botulinum Toxin Formation 260 AND that is not hermetically sealed, or an LACF, • For brine to fish ratio: or a frozen product); ° At the start of the brining process; OR AND • Destroy the product; • For time requirements of the drying process: OR ° Each batch; • Divert the product to a non-food use. AND AND • For all other critical factors specified by the Take the following corrective action to regain control study: over the operation after a critical limit deviation: ° As often as necessary to maintain control; • Adjust the salt and/or nitrite concentration in OR the brine; • For water phase salt and, when appropriate, OR nitrite: • Adjust the air velocity or input air ° Each lot or batch of finished product. temperature to the drying chamber; » Who Will Do the Monitoring? OR • For continuous temperature-recording • Extend the drying process to compensate devices: for a reduced air velocity or temperature or elevated humidity; ° Monitoring is performed by the device itself. The visual check of the data OR generated by the device, to ensure • Adjust the brine strength or brine to fish ratio; that the critical limits have been met consistently, may be performed by any OR person who has an understanding of the • Cool the brine; nature of the controls; OR OR • Move some or all of the product to another • For other checks: drying chamber; ° Any person who has an understanding of OR the nature of the controls. • Make repairs or adjustments to the drying chamber as necessary. Establish Corrective Action Procedures. Take the following corrective action to a product Establish a Recordkeeping System. involved in a critical limit deviation: • Printouts, charts, or readings from continuous • Chill and hold the product until its safety can temperature-recording devices; be evaluated; AND OR • Record of visual checks of recorded data; • Reprocess the product; AND OR • Appropriate records (e.g., processing record • Divert the product to a use in which the showing the results of the brine strength critical limit is not applicable (e.g., packaging and temperature, brine to fish ratio, size

CHAPTER 13: Clostridium botulinum Toxin Formation 261 and species of fish, and time of brining) as applicable, nitrite levels should be necessary to document the monitoring of taken into consideration in the process the critical factors of the brining, dry salting, establishment. A record of the process and/or drying process, as established by a establishment should be maintained; study; AND OR • Before a temperature-indicating device (e.g., • Results of the finished product water phase a thermometer) or temperature-recording salt determination and, when appropriate, device (e.g., a recording thermometer) is nitrite determination. put into service, check the accuracy of the device to verify that the factory calibration Establish Verification Procedures. has not been affected. This check can be • Process validation study (except where water accomplished by: phase salt analysis and, where appropriate, ° Immersing the sensor in an ice slurry nitrite analysis of the finished product are the (32°F (0°C)), if the device will be used at monitoring procedure): or near refrigeration temperature; ° The adequacy of the brining, dry OR salting, and drying processes should ° Immersing the sensor in boiling water be established by a scientific study. It (212°F (100°C)) if the device will be used should be designed to consistently at or near the boiling point. Note that achieve a water phase salt level of the temperature should be adjusted to 3.5% or 3% with not less than 100 ppm compensate for altitude, when necessary; nitrite. Expert knowledge of salting and/ or drying processes may be required OR to establish such a process. Such ° Doing a combination of the above if knowledge can be obtained by education the device will be used at or near room or experience, or both. Process temperature; validation study for establishment of OR brining, dry salting, and drying processes may require access to adequate facilities ° Comparing the temperature reading and the application of recognized on the device with the reading on a methods. The drying equipment should known accurate reference device (e.g., be designed, operated, and maintained to a thermometer traceable to National deliver the established drying process to Institute of Standards and Technology every unit of product. In some instances, (NIST) standards) under conditions that brining, dry salting, and/or drying studies are similar to how it will be used (e.g., may be required to establish minimum air temperature, brine temperature, processes. In other instances, existing product internal temperature) within the literature, which establishes minimum temperature range at which it will be processes or adequacy of equipment, used; is available. Characteristics of the AND process, product, and/or equipment • Once in service, check the temperature- that affect the ability of the established indicating device or temperature-recording minimum salting, dry salting, and drying device daily before the beginning of process to deliver the desired finished operations. Less frequent accuracy checks product water phase salt and, where may be appropriate if they are recommended

CHAPTER 13: Clostridium botulinum Toxin Formation 262 by the instrument manufacturer and the and verification records within 1 week of history of use of the instrument in your preparation to ensure they are complete and facility has shown that the instrument any critical limit deviations that occurred consistently remains accurate for a longer were appropriately addressed. period of time. In addition to checking that the device is accurate by one of the methods 1B. COLD SMOKING described above, this process should include Set Critical Limits. a visual examination of the sensor and any • The smoker temperature must not exceed attached wires for damage or kinks. The 90°F (32.2°C). device should be checked to ensure that it is operational and, where applicable, has Establish Monitoring Procedures. sufficient ink and paper; AND » What Will Be Monitored? • Calibrate the temperature-indicating device • The smoker temperature. or temperature recording device against a » How Will Monitoring Be Done? known accurate reference device (e.g., a • Measure ambient smoker chamber NIST-traceable thermometer) at least once a temperature using a continuous temperature- year or more frequently if recommended by recording device (e.g., a recording the device manufacturer. Optimal calibration thermometer). frequency is dependent upon the type, condition, past performance, and conditions » How Often Will Monitoring Be Done (Frequency)? of use of the device. Consistent temperature • Continuous monitoring by the device itself, variations away from the actual value (drift) with a visual check of the recorded data at found during checks and/or calibration may least once per batch. show a need for more frequent calibration or the need to replace the device (perhaps with » Who Will Do the Monitoring? a more durable device). Devices subjected • Monitoring is performed by the device itself. to high temperatures for extended periods of The visual check of the data generated time may require more frequent calibration. by the device, to ensure that the critical Calibration should be performed at a limits have been met consistently, may minimum of two temperatures that bracket be performed by any person who has an the temperature range at which it is used; understanding of the nature of the controls. AND Establish Corrective Action Procedures. • Perform other calibration procedures as necessary to ensure the accuracy of the Take the following corrective action to a product monitoring instruments; involved in a critical limit deviation: AND • Chill and hold the product until its safety can be evaluated; • Do finished product sampling and analysis to determine water phase salt and, where OR appropriate, nitrite analysis at least once • Divert the product to a use in which the every 3 months (except where such testing is critical limit is not applicable (e.g., packaging performed as part of monitoring); that is not hermetically sealed, or an LACF, AND or a frozen product); • Review monitoring, corrective action, OR

CHAPTER 13: Clostridium botulinum Toxin Formation 263 • Destroy the product; traceable thermometer) under conditions that are similar to how it will be used (e.g., OR air temperature) within the temperature • Divert the product to a non-food use. range at which it will be used; AND AND Take the following corrective action to regain control • Once in service, check the temperature- over the operation after a critical limit deviation: recording device daily before the beginning • Make repairs or adjustments to the smoking of operations. Less frequent accuracy checks chamber; may be appropriate if they are recommended by the instrument manufacturer and the AND/OR history of use of the instrument in your • Move some or all of the product to another facility has shown that the instrument smoking chamber. consistently remains accurate for a longer period of time. In addition to checking that Establish a Recordkeeping System. the device is accurate by one of the methods • Printouts, charts, or readings from continuous described above, this process should include temperature-recording devices; a visual examination of the sensor and any attached wires for damage or kinks. The AND device should be checked to ensure that it • Record of visual checks of recorded data. is operational and, where applicable, has sufficient ink and paper; Establish Verification Procedures. AND • Before a temperature-recording device (e.g., a recording thermometer) is put into service, • Calibrate the temperature-recording device check the accuracy of the device to verify that against a known accurate reference device the factory calibration has not been affected. (e.g., a NIST-traceable thermometer) at This check can be accomplished by: least once a year or more frequently if recommended by the device manufacturer. Immersing the sensor in an ice slurry ° Optimal calibration frequency is dependent (32°F (0°C)) if the device will be used at upon the type, condition, past performance, or near refrigeration temperature; and conditions of use of the device. OR Consistent temperature variations away from ° Immersing the sensor in boiling water the actual value (drift) found during checks (212°F (100°C)) if the device will be used and/or calibration may show a need for more at or near the boiling point. Note that frequent calibration or the need to replace the temperature should be adjusted to the device (perhaps with a more durable compensate for altitude, when necessary; device). Calibration should be performed at a minimum of two temperatures that bracket OR the temperature range at which it is used; ° Doing a combination of the above if the device will be used at or near room AND temperature; • Review monitoring, corrective action, and verification records within 1 week of OR preparation to ensure they are complete and ° Comparing the temperature reading on any critical limit deviations that occurred the device with the reading on a known were appropriately addressed. accurate reference device (e.g., a NIST-

CHAPTER 13: Clostridium botulinum Toxin Formation 264 1C. HOT SMOKING OR Set Critical Limits. • Destroy the product; • The internal temperature of the fish must OR be maintained at or above 145°F (62.8°C) • Divert the product to a non-food use. throughout the fish for at least 30 minutes. AND

Establish Monitoring Procedures. Take the following corrective action to regain control over the operation after a critical limit deviation: » What Will Be Monitored? • Make repairs or adjustments to the heating • The internal temperature at the thickest chamber; portion of three of the largest fish in the OR smoking chamber. • Move some or all of the product to another » How Will Monitoring Be Done? heating chamber. • Use a continuous temperature-recording device (e.g., a recording thermometer) Establish a Recordkeeping System. equipped with three temperature-sensing • Printouts, charts, or readings from continuous probes. temperature-recording devices; » How Often Will Monitoring Be Done (Frequency)? AND • Continuous monitoring by the device itself, • Record of visual checks of recorded data. with visual check of the recorded data at least once per batch. Establish Verification Procedures. • Before a temperature-recording device (e.g., » Who Will Do the Monitoring? a recording thermometer) is put into service,

• Monitoring is performed by the device itself. check the accuracy of the device to verify The visual check of the data generated that the factory calibration has not been by the device, to ensure that the critical affected. This check can be accomplished limits have been met consistently, may by: be performed by any person who has an Immersing the sensor in an ice slurry understanding of the nature of the controls. ° (32°F (0°C)) if the device will be used at or near refrigeration temperature; Establish Corrective Action Procedures. OR Take the following corrective action to a product involved in a critical limit deviation: ° Immersing the sensor in boiling water (212°F (100°C)) if the device will be used • Chill and hold the product until its safety can at or near the boiling point. Note that be evaluated; the temperature should be adjusted to OR compensate for altitude, when necessary; • Reprocess the product; OR OR ° Doing a combination of the above if • Divert the product to a use in which the the device will be used at or near room critical limit is not applicable (e.g., packaging temperature; that is not hermetically sealed, or a LACF, or OR a frozen product);

CHAPTER 13: Clostridium botulinum Toxin Formation 265 ° Comparing the temperature reading on preparation to ensure they are complete and the device with the reading on a known any critical limit deviations that occurred accurate reference device (e.g., a NIST- were appropriately addressed. traceable thermometer) under conditions that are similar to how it will be used (e.g., 1D. REFRIGERATED FINISHED PRODUCT STORAGE product internal temperature) within the Set Critical Limits. temperature range at which it will be used; • For refrigerated (not frozen) finished product AND storage: • Once in service, check the temperature- ° The product is held at a cooler recording device daily before the beginning temperature of 40°F (4.4°C) or of operations. Less frequent accuracy checks below. Note that allowance for routine may be appropriate if they are recommended refrigeration defrost cycles may be by the instrument manufacturer and the necessary. Also note that you may history of use of the instrument in your choose to set a critical limit that specifies facility has shown that the instrument a time and temperature of exposure to consistently remains accurate for a longer temperatures above 40°F (4.4°C); period of time. In addition to checking that OR the device is accurate by one of the methods described above, this process should include • For finished product stored under ice: a visual examination of the sensor and any ° The product is completely and attached wires for damage or kinks. The continuously surrounded by ice device should be checked to ensure that it throughout the storage time. is operational and, where applicable, has sufficient ink and paper; Establish Monitoring Procedures.

AND » What Will Be Monitored? • Calibrate the temperature-recording device • For refrigerated finished product storage: against a known accurate reference device The temperature of the cooler; (e.g., a NIST-traceable thermometer) at ° least once a year or more frequently if OR recommended by the device manufacturer. • For finished product storage under ice: Optimal calibration frequency is dependent ° The adequacy of ice surrounding the upon the type, condition, past performance, product. and conditions of use of the device. Consistent temperature variations away from » How Will Monitoring Be Done? the actual value (drift) found during checks • For refrigerated finished product storage: and/or calibration may show a need for more ° Use a continuous temperature-recording frequent calibration or the need to replace device (e.g., a recording thermometer); the device (perhaps with a more durable device). Calibration should be performed at OR a minimum of two temperatures that bracket • For finished product storage under ice: the temperature range at which it is used; ° Make visual observations of the AND adequacy of ice in a representative number of containers (e.g., cartons and • Review monitoring, corrective action, totes) from throughout the cooler. and verification records within 1 week of

CHAPTER 13: Clostridium botulinum Toxin Formation 266 » How Often Will Monitoring Be Done (Frequency)? OR • For continuous temperature-recording ° Move some or all of the product in the devices: malfunctioning cooler to another cooler; ° Continuous monitoring by the device OR itself, with a visual check of the recorded Freeze the product; data at least once per day; ° AND OR • Address the root cause: • For finished product storage under ice: ° Make repairs or adjustments to the ° Sufficient frequency to ensure control. malfunctioning cooler; » Who Will Do the Monitoring? OR • For continuous temperature-recording ° Make adjustments to the ice application devices: operations. ° Monitoring is performed by the device itself. The visual check of the data Establish a Recordkeeping System. generated by the device, to ensure • For refrigerated finished product storage: that the critical limits have been met Printouts, charts, or readings from consistently, may be performed by any ° continuous temperature-recording person who has an understanding of the devices; nature of the controls; AND OR Record of visual checks of recorded data; • For other checks: ° ° Any person who has an understanding of OR the nature of the controls. • For finished product storage under ice: ° Results of ice checks: Establish Corrective Action Procedures. • The number of containers examined Take the following corrective action to a product and the sufficiency of ice for each; involved in a critical limit deviation: AND • Chill and hold the affected product until an evaluation of the total time and temperature • The approximate number of exposure is performed; containers in the cooler.

OR Establish Verification Procedures. • Destroy the product; • Before a temperature-recording device (e.g., OR a recording thermometer) is put into service, • Divert the product to a non-food use. check the accuracy of the device to verify that the factory calibration has not been affected. AND This check can be accomplished by: Take the following corrective actions to regain control ° Immersing the sensor in an ice slurry over the operation after a critical limit deviation: (32°F (0°C)) if the device will be used at • Prevent further deterioration: or near refrigeration temperature; ° Add ice to the product; OR

CHAPTER 13: Clostridium botulinum Toxin Formation 267 ° Comparing the temperature reading on of fish to ensure that the ice is sufficient the device with the reading on a known to maintain product temperatures at 40°F accurate reference device (e.g., a NIST- (4.4°C) or less; traceable thermometer) under conditions AND that are similar to how it will be used (e.g., air temperature) within the temperature • Review monitoring, corrective action, range at which it will be used; and verification records within 1 week of preparation to ensure they are complete and AND any critical limit deviations that occurred • Once in service, check the temperature- were appropriately addressed. recording device daily before the beginning of operations. Less frequent accuracy checks 1E. RECEIPT OF PRODUCTS BY SECONDARY may be appropriate if they are recommended PROCESSOR by the instrument manufacturer and the Set Critical Limits. history of use of the instrument in your • For fish or fishery products delivered facility has shown that the instrument refrigerated (not frozen): consistently remains accurate for a longer period of time. In addition to checking that ° All lots received are accompanied by the device is accurate by one of the methods transportation records that show that described above, this process should include the product was held at or below 40°F a visual examination of the sensor and any (4.4°C) throughout transit. Note that attached wires for damage or kinks. The allowance for routine refrigeration device should be checked to ensure that it defrost cycles may be necessary; is operational and, where applicable, has OR sufficient ink and paper; • For products delivered under ice: AND ° Product is completely surrounded by ice • Calibrate the temperature-recording device at the time of delivery; against a known accurate reference device OR (e.g., a NIST-traceable thermometer) at • For products delivered under chemical least once a year or more frequently if cooling media, such as gel packs: recommended by the device manufacturer. Optimal calibration frequency is dependent ° There is an adequate quantity of cooling upon the type, condition, past performance, media that remain frozen to have and conditions of use of the device. maintained product at 40°F (4.4°C) or Consistent temperature variations away from below throughout transit; the actual value (drift) found during checks AND and/or calibration may show a need for more The internal temperature of the product frequent calibration or the need to replace ° at the time of delivery is 40°F (4.4°C) or the device (perhaps with a more durable below; device). Calibration should be performed at a minimum of two temperatures that bracket OR the temperature range at which it is used; • For products delivered refrigerated (not AND frozen) with a transit time (including all time outside a controlled temperature • When visual checks of ice are used, environment) of 4 hours or less (optional periodically measure internal temperatures control strategy):

CHAPTER 13: Clostridium botulinum Toxin Formation 268 ° Time of transit does not exceed 4 hours; containers (e.g., cartons and totes) at the time of delivery. AND ° Temperature of the product at the time » How Will Monitoring Be Done? of delivery does not exceed 40°F (4.4°C). • For products delivered refrigerated (not Note: Processors receiving product with transit times of 4 hours or less frozen): may elect to use one of the controls described for longer transit times. ° Use a continuous temperature-recording device (e.g., a recording thermometer) Establish Monitoring Procedures. for internal product temperature or ambient air temperature monitoring » What Will Be Monitored? during transit; • For products delivered refrigerated (not frozen): OR ° The internal temperature of the product • For products delivered under ice: throughout transportation; ° Make visual observations of the OR adequacy of ice in a representative number of containers (e.g., cartons and The temperature within the truck or ° totes) from throughout the shipment, at other carrier throughout transportation; delivery; OR OR • For products delivered under ice: • For products delivered under chemical ° The adequacy of ice surrounding the cooling media, such as gel packs: product at the time of delivery; ° Make visual observations of the OR adequacy and frozen state of the cooling • For products held under chemical cooling media in a representative number of media, such as gel packs: containers (e.g., cartons and totes) from throughout the shipment, at delivery; ° The quantity and frozen status of cooling media at the time of delivery; AND AND ° Use a temperature-indicating device (e.g., a thermometer) to determine internal ° The internal temperature of a representative number of product product temperatures in a representative containers (e.g., cartons and totes) at number of product containers from time of delivery; throughout the shipment, at delivery; OR OR • For products delivered refrigerated (not • For products delivered refrigerated (not frozen) with a transit time of 4 hours or less: frozen) with a transit time of 4 hours or less: ° The date and time fish were removed ° Review carrier records to determine from a controlled temperature the date and time the product was environment before shipment and the removed from a controlled temperature date and time delivered; environment before shipment and the date and time delivered; AND AND ° The internal temperature of a representative number of product

CHAPTER 13: Clostridium botulinum Toxin Formation 269 ° Use a temperature-indicating device (e.g., • Discontinue use of the supplier or carrier a thermometer) to determine internal until evidence is obtained that the identified product temperatures in a representative transportation-handling practices have been number of product containers (e.g., improved. cartons and totes) randomly selected from throughout the shipment, at Establish a Recordkeeping System. delivery. Measure a minimum of 12 • Receiving records showing: product containers, unless there are Results of continuous temperature fewer than 12 product containers in a ° monitoring: lot, in which case measure all of the containers. Lots that show a high level • Printouts, charts, or readings of temperature variability may require a from continuous temperature- larger sample size. recording devices;

» How Often Will Monitoring Be Done (Frequency)? AND • Each lot received. • Visual check of recorded data; OR » Who Will Do the Monitoring? Results of ice checks, including: • For continuous temperature-recording ° devices: • The number of containers examined ° Monitoring is performed by the device and the sufficiency of ice for each; itself. The visual check of the data AND generated by the device, to ensure • The number of containers in the lot; that the critical limits have been met consistently, may be performed by any OR person who has an understanding of the ° Results of the chemical media checks, nature of the controls; including: OR • The number of containers • For other checks: examined and the frozen status of the media for each; ° Any person who has an understanding of the nature of the controls. AND • The number of containers in the lot; Establish Corrective Action Procedures. AND/OR Take the following corrective action to a product ° Results of internal product temperature involved in a critical limit deviation: monitoring, including: • Chill and hold the affected product until an • The number of containers evaluation of the total time and temperature examined and the internal exposure is performed; temperatures observed for each; OR AND • Reject the lot. • The number of containers in the lot; AND AND Take the following corrective action to regain control • Date and time fish were initially over the operation after a critical limit deviation: removed from a controlled

CHAPTER 13: Clostridium botulinum Toxin Formation 270 temperature environment recommended by the device manufacturer. and date and time fish were Optimal calibration frequency is dependent delivered, when applicable. upon the type, condition, past performance, and conditions of use of the device. Establish Verification Procedures. Consistent temperature variations away from • Before a temperature-indicating device (e.g., the actual value (drift) found during checks a thermometer) is put into service, check and/or calibration may show a need for more the accuracy of the device to verify that the frequent calibration or the need to replace factory calibration has not been affected. the device (perhaps with a more durable This check can be accomplished by: device). Calibration should be performed at ° Immersing the sensor in an ice slurry a minimum of two temperatures that bracket (32°F (0°C)), if the device will be used at the temperature range at which it is used; or near refrigeration temperature; AND OR • Check the accuracy of temperature-recording ° Comparing the temperature reading on devices that are used for monitoring transit the device with the reading on a known conditions, for all new suppliers and at accurate reference device (e.g., a NIST- least quarterly for each supplier thereafter. traceable thermometer) under conditions Additional checks may be warranted based that are similar to how it will be used on observations at receipt (e.g., refrigeration (e.g., product internal temperature) units appear to be in poor repair or readings within the temperature range at which it appear to be erroneous). The accuracy of will be used; the device can be checked by comparing the temperature reading on the device with AND the reading on a known accurate reference • Once in service, check the temperature- device (e.g., a NIST-traceable thermometer) indicating device daily before the under conditions that are similar to how it beginning of operations. Less frequent will be used (e.g., air temperature) within the accuracy checks may be appropriate if temperature range at which it will be used; they are recommended by the instrument manufacturer and the history of use of AND the instrument in your facility has shown • When visual checks of ice are used, that the instrument consistently remains periodically measure internal temperatures accurate for a longer period of time. In of fish to ensure that the ice or is sufficient addition to checking that the device is to maintain product temperatures at 40°F accurate by one of the methods described (4.4°C) or less; above, this process should include a AND visual examination of the sensor and any • Review monitoring, corrective action, attached wires for damage or kinks. The and verification records within 1 week of device should be checked to ensure that preparation to ensure they are complete and it is operational; any critical limit deviations that occurred AND were appropriately addressed. • Calibrate the temperature-indicating device against a known accurate reference device (e.g., a NIST-traceable thermometer) at least once a year or more frequently if

CHAPTER 13: Clostridium botulinum Toxin Formation 271 TABLE 13-1

CONTROL STRATEGY EXAMPLE 1 - SMOKING

This table is an example of a portion of a HACCP plan using “Control Strategy Example 1 - Smoking.” This example illustrates how a processor of vacuum-packaged hot- smoked salmon can control C. botulinum toxin formation. It is provided for illustrative purposes only.

C. botulinum toxin formation may be only one of several significant hazards for this product. Refer to Tables 3-2 and 3-4 (Chapter 3) for other potential hazards (e.g., aquaculture drugs, environmental chemical contaminants and pesticides, parasites, growth of other pathogenic bacteria, survival of other pathogenic bacteria through the cook step, and metal fragments).

Example Only See Text for Full Recommendations

CHAPTER 13: ToxinCHAPTER Formation botulinum Clostridium (1) (2) (3) (4) (5) (6) (7) (8) (9) (10)

CRITICAL LIMITS MONITORING CRITICAL SIGNIFICANT FOR EACH CORRECTIVE CONTROL RECORDS VERIFICATION HAZARD(S) PREVENTIVE ACTION(S) POINT WHAT HOW FREQUENCY WHO MEASURE* Brining C. botulinum Minimum Start time Clock Every batch Brine Extend the Production Establish toxin brining time: and end time room brining record a brining and formation in 6 hours of the brining Dial Every 2 hours employee process drying process 272 the finished process thermometer product Maximum brine Hold and evaluate Check the dial thermometer for temperature: 40ºF Brine the product accuracy and damage and to temperature Cool the brine ensure that it is operational before putting into operation; Minimum salt Salt Salinometer Start of each Brine Add salt Production check it daily, at the beginning of concentration of concentration brining room record operations; and calibrate it once brine at the start of brine process employee per year of brining: 60° salinometer Monthly calibration of the scale Minimum ratio of Weight of Visual, to Start of each Brine Add brine Production brine to fish: brine (as mark on the brining room record Quarterly water phase salt analysis 2:1 determined tank process employee of the finished product by volume) Review monitoring, corrective action, and verification records Weight of Scale Each batch Remove some fish within 1 week of preparation fish and reweigh Maximum fish Fish Caliper Each batch Brine Hold and evaluate Production thickness 1½ in. thickness (10 largest room based on finished record Note: To produce fish) employee product water a minimum water phase salt analysis phase salt level in the loin muscle of 3.5% TABLE 13-1

CONTROL STRATEGY EXAMPLE 1 - SMOKING

This table is an example of a portion of a HACCP plan using “Control Strategy Example 1 - Smoking.” This example illustrates how a processor of vacuum-packaged hot- smoked salmon can control C. botulinum toxin formation. It is provided for illustrative purposes only.

C. botulinum toxin formation may be only one of several significant hazards for this product. Refer to Tables 3-2 and 3-4 (Chapter 3) for other potential hazards (e.g., aquaculture drugs, environmental chemical contaminants and pesticides, parasites, growth of other pathogenic bacteria, survival of other pathogenic bacteria through the cook step, and metal fragments).

Example Only See Text for Full Recommendations

(1) (2) (3) (4) (5) (6) (7) (8) (9) (10) CHAPTER 13: ToxinCHAPTER Formation botulinum Clostridium

CRITICAL LIMITS MONITORING CRITICAL SIGNIFICANT FOR EACH CORRECTIVE CONTROL RECORDS VERIFICATION HAZARD(S) PREVENTIVE ACTION(S) POINT WHAT HOW FREQUENCY WHO MEASURE* Smoking C. botulinum Minimum time Time of open Clock Each batch Smoker Extend the drying Production Establish a brining and drying and toxin open vent: 2 hours vent employee process record process Quarterly water phase salt drying formation Hold and evaluate analysis of the finished product

273 in finished based on finished Review monitoring, corrective product product water action, and verification records phase salt analysis within 1 week of preparation Heating C. botulinum Internal Internal Digital data Continuous, Smoker Extend the Data Check the data logger for accuracy toxin temperature of fish temperature logger with with visual employee heating logger and damage and to ensure that formation in held at or above of fish and three probes check of process printout it is operational before putting the finished 145°F for at least 30 time at that in thickest recorded data Make repairs or into operation; check it daily, at product minutes temperature fish in cold at the end of adjustments to the the beginning of operations; and spot of the batch smoking chamber calibrate it once per year smoking Review monitoring, corrective chamber Hold and evaluate action, and verification records the product within 1 week of preparation

Finished C. botulinum Maximum cooler Cooler air Digital data Continuous, Production Adjust or repair Digital Check the data logger for accuracy product toxin temperature: 40°F temperature logger with visual employee the cooler data logger and damage and to ensure that storage formation (based on growth check of Hold and evaluate printout it is operational before putting during of vegetative recorded data the product based into operation; check it daily, at finished pathogens) once per day on time and the beginning of operations; and product temperature of calibrate it once per year storage exposure Review monitoring, corrective action, and verification records within 1 week of preparation

*Note: The critical limits in this example are for illustrative purposes only and are not related to any recommended process. • CONTROL STRATEGY EXAMPLE 2 - ° Performance data from the manufacturer; REFRIGERATION WITH TTI AND This control strategy should include the following • For transportation conditions: elements, as appropriate: ° The temperature within the truck or a. Unactivated TTI receipt; other carrier throughout transportation;

b. Unactivated TTI storage; OR ° Other conditions that affect the c. Application and activation of TTI; functionality of the TTI, where applicable; d. Refrigerated finished product storage; AND e. Receipt of product by secondary • For functionality at receipt: processor. ° The ability of the TTI to produce an 2A. UNACTIVATED TTI RECEIPT alert indicator, such as a color change of the device, when exposed to time and Set Critical Limits. temperature abuse at time of receipt. • The TTI is suitable for use. It should be designed to perform properly under the » How Will Monitoring Be Done? conditions that it will be used. It should • For suitability of use: also be designed to produce an alert ° Review performance data; indicator (e.g., a color change of the device) AND at a combination of time and temperature exposures that will prevent the formation of • For transportation conditions: non-proteolytic C. botulinum toxin formation ° Use a continuous temperature-recording (e.g., consistent with the “Skinner-Larkin device (e.g., a recording thermometer) curve”); for ambient air temperature monitoring during transit; AND • Where transportation conditions (e.g., AND temperature) could affect the functionality • For functionality at receipt: of the TTI, all lots of TTIs are accompanied ° Activate and then expose a TTI from by transportation records that show that they the lot to ambient air temperature for were held at conditions that do not result in sufficient time to determine whether loss of functionality throughout transit; it is functional (i.e., produces an alert AND indicator, such as a color change of the device). • The TTI functions (i.e., produces an alert indicator, such as a color change of » How Often Will Monitoring Be Done (Frequency)? the device, when exposed to time and • For suitability of use: temperature abuse) at time of receipt. ° The first shipment of a TTI model; Establish Monitoring Procedures. AND • For transportation conditions and » What Will Be Monitored? functionality at receipt: • For suitability of use: ° Every shipment.

CHAPTER 13: Clostridium botulinum Toxin Formation 274 » Who Will Do the Monitoring? ° Records of visual checks of recorded • For suitability of use: data; ° Anyone with an understanding of TTI AND validation studies and of the intended • For functionality at receipt: conditions of use; ° Results of a TTI challenge test (i.e., AND whether the TTI produces an alert • For transportation conditions and indicator, such as a color change of functionality at receipt: the device, when exposed to time and temperature abuse). ° Anyone with an understanding of the nature of the controls. Establish Verification Procedures. Establish Corrective Action Procedures. • Check the accuracy of temperature-recording devices that are used for monitoring transit Take the following corrective action to a product conditions, for all new suppliers and at involved in a critical limit deviation: least quarterly for each supplier thereafter. • Reject or return the shipment. Additional checks may be warranted based AND on observations at receipt (e.g., refrigeration units appear to be in poor repair or readings Take the following corrective actions to regain control appear to be erroneous). The accuracy of over the operation after a critical limit deviation: the device can be checked by comparing • For suitability of use: the temperature reading on the device with ° Discontinue use of the supplier until the reading on a known accurate reference documentation of validation has been device (e.g., a NIST-traceable thermometer) provided; under conditions that are similar to how it will be used (e.g., air temperature) within the AND temperature range at which it will be used; • For transportation conditions and functionality at receipt: AND • Review monitoring, corrective action, ° Discontinue use of the supplier or carrier until evidence is obtained that the and verification records within 1 week of identified production or transportation preparation to ensure they are complete and practices have been improved. any critical limit deviations that occurred were appropriately addressed.

Establish a Recordkeeping System. 2B. UNACTIVATED TTI STORAGE

• For suitability of use: Set Critical Limits. Manufacturer’s performance data; ° • The combination of storage conditions AND (e.g., temperature) that prevent loss of • For transportation conditions: functionality throughout storage (based ° Printouts, charts, or readings from on manufacturer’s specifications). continuous temperature-recording devices; AND

CHAPTER 13: Clostridium botulinum Toxin Formation 275 Establish Monitoring Procedures. Establish Corrective Action Procedures.

» What Will Be Monitored? Take the following corrective action to a TTI involved in a critical limit deviation: • Storage air temperature, where temperature affects functionality of the TTI; • Destroy the lot of TTIs. AND/OR AND • Other storage conditions that affect Take the following corrective action to regain control functionality of the TTI. over the operation after a critical limit deviation: • Make repairs or adjustments to the » How Will Monitoring Be Done? malfunctioning cooler; • For temperature: AND/OR ° Use a continuous temperature-recording device (e.g., a recording thermometer); • Make other repairs or adjustment appropriate for the condition. AND/OR • For other conditions: Establish a Recordkeeping System. ° Use instruments appropriate for the • For refrigerated storage: purpose. ° Printouts, charts, or readings from » How Often Will Monitoring Be Done (Frequency)? continuous temperature-recording devices; • For temperature: ° Continuous monitoring by the device AND itself, with a visual check of the recorded ° Record of visual checks of recorded data; data at least once per day; AND/OR AND/OR • Storage record showing the results of • For other conditions: monitoring of other conditions. ° With sufficient frequency to ensure control. Establish Verification Procedures. • Before a temperature-recording device (e.g., Who Will Do the Monitoring? » a recording thermometer) is put into service, • With continuous temperature-recording check the accuracy of the device to verify that devices: the factory calibration has not been affected. ° Monitoring is performed by the device This check can be accomplished by: itself. The visual check of the data ° Immersing the sensor in an ice slurry generated by the device, to ensure (32°F (0°C)) if the device will be used at that the critical limits have been met or near refrigeration temperature; consistently, may be performed by any person who has an understanding of the OR nature of the controls; ° Comparing the temperature reading on the device with the reading on a known AND accurate reference device (e.g., a NIST- • For other checks: traceable thermometer) under conditions ° Any person who has an understanding of that are similar to how it will be used (e.g., the nature of the controls. air temperature) within the temperature range at which it will be used;

CHAPTER 13: Clostridium botulinum Toxin Formation 276 AND 2C. APPLICATION AND ACTIVATION OF TTI • Once in service, check the temperature- Set Critical Limits. recording device daily before the beginning of operations. Less frequent accuracy checks • Each consumer package has an activated may be appropriate if they are recommended TTI. by the instrument manufacturer and the history of use of the instrument in your Establish Monitoring Procedures. facility has shown that the instrument » What Will Be Monitored? consistently remains accurate for a longer • Packages for the presence of an activated period of time. In addition to checking that TTI. the device is accurate by one of the methods described above, this process should include » How Will Monitoring Be Done? a visual examination of the sensor and any • Visual examination. attached wires for damage or kinks. The device should be checked to ensure that it » How Often Will Monitoring Be Done (Frequency)? is operational and, where applicable, has • Representative number of packages from sufficient ink and paper; each lot of product. AND » Who Will Do the Monitoring? • Calibrate the temperature-recording device • Any person who has an understanding of the against a known accurate reference device nature of the controls. (e.g., a NIST-traceable thermometer) at least once a year or more frequently if recommended by the device manufacturer. Establish Corrective Action Procedures. Optimal calibration frequency is dependent Take the following corrective action to a product upon the type, condition, past performance, involved in a critical limit deviation: and conditions of use of the device. • Hold the lot below 38°F (3.3°C) until TTIs Consistent temperature variations away from are applied and activated. the actual value (drift) found during checks and/or calibration may show a need for more AND frequent calibration or the need to replace Take the following corrective action to regain control the device (perhaps with a more durable over the operation after a critical limit deviation: device). Calibration should be performed at • Identify and correct the cause of the TTI a minimum of two temperatures that bracket application or activation deficiency. the temperature range at which it is used; AND Establish a Recordkeeping System. • Perform other instrument calibration, as • Packaging control record that shows the appropriate; results of the TTI checks. AND Establish Verification Procedures. • Review monitoring, corrective action, • Review monitoring and corrective action and verification records within 1 week of records within 1 week of preparation preparation to ensure they are complete and to ensure they are complete and any any critical limit deviations that occurred critical limit deviations that occurred were were appropriately addressed. appropriately addressed.

CHAPTER 13: Clostridium botulinum Toxin Formation 277 2D. REFRIGERATED FINISHED PRODUCT STORAGE Follow the guidance for “Control Strategy Example 1 - Smoking (1d - Refrigerated Finished Product Storage),” except that the where the critical limits list 40ºF (4.4ºC), they should list 38°F (3.3°C).

2E. RECEIPT OF PRODUCTS BY SECONDARY PROCESSOR Follow the guidance for “Control Strategy Example 1 - Smoking (1e - Receipt of Products by Secondary Processor),” except that the where the critical limits list 40ºF (4.4ºC), they should list 38°F (3.3°C).

CHAPTER 13: Clostridium botulinum Toxin Formation 278 TABLE 13-2

CONTROL STRATEGY EXAMPLE 2 - REFRIGERATION WITH TTI

This table is an example of a portion of a HACCP plan using “Control Strategy Example 2 - Refrigeration With TTI.” This example illustrates how a processor of refrigerated, vacuum-packaged, raw fish fillets can control C. botulinum toxin formation. It is provided for illustrative purposes only.

C. Botulinum toxin formation may be only one of several significant hazards for this product. Refer to Tables 3-2 and 3-4 (Chapter 3) for other potential hazards (e.g., aquaculture drugs, environmental chemical contaminants and pesticides, parasites, growth of other pathogenic bacteria, and metal fragments).

Example Only See Text for Full Recommendations

(1) (2) (3) (4) (5) (6) (7) (8) (9) (10) CHAPTER 13: ToxinCHAPTER Formation botulinum Clostridium CRITICAL LIMITS MONITORING CRITICAL SIGNIFICANT FOR EACH CONTROL CORRECTIVE ACTION(S) RECORDS VERIFICATION HAZARD(S) PREVENTIVE POINT WHAT HOW FREQUENCY WHO MEASURE Receipt of C. botulinum TTI is Performance Review of First Quality Reject the Manufacturer’s Review TTI toxin suitable for use data from the performance shipment of a assurance shipment performance monitoring, formation in manufacturer data TTI model supervisor Discontinue use of the data corrective the finished supplier until appropriate action records 279 product validation within 1 week documentation of preparation is provided All lots received Truck Digital time and Continuous, Receiving Discontinue use of the Receiving Check the data are temperature temperature with visual employee supplier or carrier until record logger for all accompanied data logger review and evidence is obtained new suppliers by truck evaluation of that the identifi ed and for all records temperature- transportation- handling suppliers at that show monitoring practices have been least quarterly temperature records for each improved thereafter was maintained shipment at or below Reject the Review 40°F shipment monitoring, corrective action records within 1 week of preparation The TTI The ability Expose a TTI Every Quality Discontinue use of the TTI Review functions of the TTI to from the lot shipment assurance supplier or carrier until challenge monitoring, at receipt change color to room air staff evidence is obtained that record corrective when exposed temperature for the identifi ed production action records to room air sufficient time or transportation- within 1 week temperature to determine handling practices have of preparation whether it been improved changes color TABLE 13-2

CONTROL STRATEGY EXAMPLE 2 - REFRIGERATION WITH TTI

This table is an example of a portion of a HACCP plan using “Control Strategy Example 2 - Refrigeration With TTI.” This example illustrates how a processor of refrigerated, vacuum-packaged, raw fish fillets can control C. botulinum toxin formation. It is provided for illustrative purposes only.

C. Botulinum toxin formation may be only one of several significant hazards for this product. Refer to Tables 3-2 and 3-4 (Chapter 3) for other potential hazards (e.g., aquaculture drugs, environmental chemical contaminants and pesticides, parasites, growth of other pathogenic bacteria, and metal fragments).

Example Only See Text for Full Recommendations

(1) (2) (3) (4) (5) (6) (7) (8) (9) (10)

CHAPTER 13: ToxinCHAPTER Formation botulinum Clostridium CRITICAL LIMITS MONITORING CRITICAL SIGNIFICANT FOR EACH CONTROL CORRECTIVE ACTION(S) RECORDS VERIFICATION HAZARD(S) PREVENTIVE POINT WHAT HOW FREQUENCY WHO MEASURE TTI C. botulinum Cooler Cooler Digital time and Continuous, Quality Repair or adjust cooler Data Check the storage toxin maintained temperature temperature with visual assurance logger printout data logger for formation in below 38°F data logger check of staff Destroy the lot of TTIs accuracy and the finished recorded data damage and to

280 product once per day ensure that it is operational before putting into operation; check it daily, at the beginning of operations; and calibrate it once per year

Review monitoring, corrective action, and verification records within 1 week of preparation TTI C. botulinum Each Packages for Visual Representative Production Hold lot below 38°F, and Packaging Review attachment toxin package has an the presence examination number of employee apply and activate TTIs control record monitoring, and formation in activated TTI of an packages from and corrective activation the finished activated TTI each lot of Identify and correct the action and product product cause of TTI application verification deviation records within 1 week of preparation TABLE 13-2

CONTROL STRATEGY EXAMPLE 2 - REFRIGERATION WITH TTI

This table is an example of a portion of a HACCP plan using “Control Strategy Example 2 - Refrigeration With TTI.” This example illustrates how a processor of refrigerated, vacuum-packaged, raw fish fillets can control C. botulinum toxin formation. It is provided for illustrative purposes only.

C. Botulinum toxin formation may be only one of several significant hazards for this product. Refer to Tables 3-2 and 3-4 (Chapter 3) for other potential hazards (e.g., aquaculture drugs, environmental chemical contaminants and pesticides, parasites, growth of other pathogenic bacteria, and metal fragments).

Example Only See Text for Full Recommendations

(1) (2) (3) (4) (5) (6) (7) (8) (9) (10)

CHAPTER 13: ToxinCHAPTER Formation botulinum Clostridium CRITICAL LIMITS MONITORING CRITICAL SIGNIFICANT FOR EACH CONTROL CORRECTIVE ACTION(S) RECORDS VERIFICATION HAZARD(S) PREVENTIVE POINT WHAT HOW FREQUENCY WHO MEASURE Finished C. botulinum Maximum Cooler air Digital data Continuous, Production Adjust or repair cooler Digital logger Check the product toxin cooler temperature logger with visual employee printout data logger for storage formation temperature check of Hold and accuracy and during 38°F recorded data evaluate the product damage and to

281 finished once per day based on time and ensure that it product temperature of exposure is operational storage before putting into operation; check it daily, at the beginning of operations; and calibrate it once per year

Review monitoring, corrective action, and verification records within 1 week of preparation

*Note: The critical limits in this example are for illustrative purposes only and are not related to any recommended process. • CONTROL STRATEGY EXAMPLE 3 - FROZEN WITH Establish Verification Procedures. LABELING • Review monitoring and corrective action Set Critical Limits. records within 1 week of preparation to ensure they are complete and any • All finished product labels must contain a critical limit deviations that occurred were “keep frozen” statement (e.g., “Important, appropriately addressed. keep frozen until used, thaw under refrigeration immediately before use”).

Establish Monitoring Procedures.

» What Will Be Monitored? • Finished product labels for the presence of a “keep frozen” statement.

» How Will Monitoring Be Done? • Visual examination.

» How Often Will Monitoring Be Done (Frequency)? • Representative number of packages from each lot of product.

» Who Will Do the Monitoring? • Any person who has an understanding of the nature of the controls.

Establish Corrective Action Procedures. Take the following corrective action to a product involved in a critical limit deviation: • Segregate and relabel any improperly labeled product. AND Take the following corrective actions to regain control over the operation after a critical limit deviation: • Segregate and return or destroy any label stock or pre-labeled packaging stock that does not contain the proper statement; AND • Determine and correct the cause of improper labels.

Establish a Recordkeeping System. • Record of labeling checks.

CHAPTER 13: Clostridium botulinum Toxin Formation 282 TABLE 13-3

CONTROL STRATEGY EXAMPLE 3 - FROZEN WITH LABELING

This table is an example of a portion of a HACCP plan using “Control Strategy Example 3 - Frozen With Labeling.” This example illustrates how a processor of frozen, vacuum-packaged, raw fish fillets can control C. botulinum toxin formation. It is provided for illustrative purposes only.

C. Botulinum toxin formation may be only one of several significant hazards for this product. Refer to Tables 3-2 and 3-4 (Chapter 3) for other potential hazards (e.g., environmental chemical contaminants and pesticides, parasites, and metal fragments).

Example Only See Text for Full Recommendations CHAPTER 13: ToxinCHAPTER Formation botulinum Clostridium

(1) (2) (3) (4) (5) (6) (7) (8) (9) (10)

MONITORING CRITICAL LIMITS CRITICAL SIGNIFICANT FOR EACH CORRECTIVE CONTROL RECORDS VERIFICATION HAZARD(S) PREVENTIVE ACTION(S) POINT WHAT HOW FREQUENCY WHO MEASURE

283 Receipt of C. botulinum All finished Finished Visual Representative Receiving Segregate and Label receiving Review labeling toxin product labels product examination number of employee relabel any record monitoring and formation must contain a labels for the packages from improperly correction during “keep presence of a each lot of labeled product action records finished frozen” “keep frozen” product within 1 week product statement statement Segregate and of preparation storage destroy any label stock that does not contain the proper statement

Determine and correct the cause of improper labels • CONTROL STRATEGY EXAMPLE 4 - PICKLING For unrefrigerated (shelf-stable), reduced oxygen- AND SALTING packaged products: This control strategy should include the following ° A water phase salt level of at least 20% elements, as appropriate: (based on the maximum salt level for growth of S. aureus); a. Brining, pickling, salting, and formulation; OR A pH of 4.6 or below; b. Refrigerated finished product storage; ° OR c. Receipt of Product by secondary ° A water activity of 0.85 or below (based processor. on the minimum water activity for growth and toxin formation of S. aureus). 4A. BRINING, PICKLING, SALTING, AND FORMULATION A heat treatment, addition of chemical additives, or other treatment may be necessary to inhibit or Set Critical Limits. eliminate spoilage organisms (e.g., mold) in shelf- • The minimum or maximum values for stable products. the critical factors of the brining, pickling, or formulation process established by a Establish Monitoring Procedures. scientific study. The critical factors are those that are necessary to ensure that the finished » What Will Be Monitored? product has: • The critical factors of the established pickling, brining, or formulation process. For refrigerated, reduced oxygen-packaged These may include: brine and acid strength; fishery products: brine or acid to fish ratio; brining and ° A water phase salt level of at least 5%; pickling time; brine and acid temperature; OR thickness, texture, fat content, quality, and ° A pH of 5.0 or below; species of fish; OR OR A water activity of below 0.97; • The water phase salt, pH, and/or water ° activity of the finished product. OR ° A water phase salt level of at least » How Will Monitoring Be Done? 2.4% in surimi-based products, when • For brine strength: combined with a pasteurization process ° Use a salinometer; in the finished product container AND of 185°F (85°C) for 15 minutes (pasteurization controls are covered in • For acid strength: Chapter 16); ° Use a pH meter or titrate for acid concentration; OR ° A combination of water phase salt, AND pH, and/or water activity that, when • For brine/acid temperature: combined, have been demonstrated to ° Use a temperature-indicating device (e.g., prevent the growth of C. botulinum type a thermometer); E and non-proteolytic types B and F. AND

CHAPTER 13: Clostridium botulinum Toxin Formation 284 • For all other critical factors specified by the Establish Corrective Action Procedures. study: Take the following corrective action to a product Use equipment appropriate for the ° involved in a critical limit deviation: measurement; • Chill and hold the product until it can be OR evaluated based on its water phase salt, pH, • For water phase salt, pH, and/or water and/or water activity level; activity: OR Collect a representative sample of the ° • Reprocess the product (if reprocessing does finished product, and conduct water not jeopardize the safety of the product); phase salt, pH, and/or water activity analysis, as appropriate. OR • Divert the product to a use in which the » How Often Will Monitoring Be Done (Frequency)? critical limit is not applicable (e.g., packaging • For brine and acid strength: that is not hermetically sealed, or a LACF, or ° At the start of each brining, pickling, and a frozen product); formulation process; OR AND • Divert the product to a non-food use; • For brine and acid temperature: OR At the start of each brining, pickling, and ° • Destroy the product. formulation process and at least every 2 hours thereafter; AND AND Take the following corrective action to regain control • For brine or acid to fish ratio: over the operation after a critical limit deviation: ° At the start of each brining, pickling, and • Adjust the brine or acid strength or brine or formulation process; acid to fish ratio; AND OR • For other critical factors specified by the study: • Extend the brining or pickling time to compensate for an improper brine or acid As often as necessary to maintain control; ° temperature. OR • Water phase salt, pH, and/or water activity Establish a Recordkeeping System. analysis should be determined for each batch • Records, as necessary, to document the of finished product. monitoring of the critical factors of the brining or pickling process, as established » Who Will Do the Monitoring? by a study (e.g., a processing record showing • For water activity: the results of the brine or acid strength ° Any person with sufficient training to and temperature, brine or acid to fish ratio, perform the analysis; size and species of fish, time of brining or pickling); OR • For other checks: OR ° Any person with an understanding of the • Record of determinations of the finished nature of the controls. product water phase salt, pH, or water activity.

CHAPTER 13: Clostridium botulinum Toxin Formation 285 Establish Verification Procedures. and/or product that affect the ability • Process validation study (except where water of the established minimum pickling, phase salt, pH, or water activity analysis brining, and formulation process of the finished product is the monitoring should be taken into consideration in procedure): the process establishment. A record of the process establishment should be The adequacy of the pickling, brining, ° maintained; and formulation process steps should be established by a scientific study. For AND refrigerated, reduced oxygen-packaged • Before a temperature-indicating device (e.g., products, it should be designed to a thermometer) is put into service, check consistently achieve: a water phase salt the accuracy of the device to verify that the level of at least 5%; a pH of 5.0 or below; factory calibration has not been affected. a water activity of below 0.97; a water This check can be accomplished by: phase salt level of at least 2.4% in surimi­ ° Immersing the sensor in an ice slurry based products, when combined with (32°F (0°C)) if the device will be used at a pasteurization process in the finished or near refrigeration temperature; product container of 185°F (85°C) for at least 15 minutes; or a combination OR of salt, pH, and/or water activity that, ° Immersing the sensor in boiling water when combined, prevent the growth of (212°F (100°C)) if the device will be C. botulinum type E and non-proteolytic used at or near the boiling point. Note types B and F (established by a scientific that the temperature should be adjusted study). For unrefrigerated (shelf-stable), to compensate for altitude, when reduced oxygen-packaged products, necessary); it should be designed to consistently OR achieve: a water phase salt level of Doing a combination of the above if at least 20% (based on the maximum ° the device will be used at or near room water phase salt level for the growth of temperature; S. aureus); a pH of 4.6 or below; or a water activity of 0.85 or below (based OR on the minimum water activity for the ° Comparing the temperature reading on growth of S. aureus). Expert knowledge the device with the reading on a known of pickling, brining, and formulation accurate reference device (e.g., a NIST- processes may be required to establish traceable thermometer) under conditions such a process. Such knowledge can be that are similar to how it will be used obtained by education or experience, or (e.g., brine temperature) within the both. Establishment of pickling, brining, temperature range at which it will be and formulation processes may require used; access to adequate facilities and the application of recognized methods. In AND some instances, pickling, brining, and • Once in service, check the temperature- formulation studies may be required to indicating device daily before the beginning establish minimum processes. In other of operations. Less frequent accuracy checks instances, existing literature, which may be appropriate if they are recommended establishes minimum processes, is by the instrument manufacturer and the available. Characteristics of the process history of use of the instrument in your

CHAPTER 13: Clostridium botulinum Toxin Formation 286 facility has shown that the instrument any critical limit deviations that occurred consistently remains accurate for a longer were appropriately addressed. period of time. In addition to checking that the device is accurate by one of the methods 4B. REFRIGERATED FINISHED PRODUCT STORAGE described above, this process should include Follow the guidance for “Control Strategy a visual examination of the sensor and any Example 1 - Smoking (1d - Refrigerated Finished attached wires for damage or kinks. The Product Storage).” device should be checked to ensure that it is 4C. RECEIPT OF PRODUCT BY SECONDARY operational; PROCESSOR AND Follow the guidance for “Control Strategy • Calibrate the temperature-indicating device Example 1 - Smoking (1e - Receipt of Product by against a known accurate reference device Secondary Processor).” (e.g., a NIST-traceable thermometer) at least once a year or more frequently if recommended by the device manufacturer. Optimal calibration frequency is dependent upon the type, condition, past performance, and conditions of use of the device. Consistent temperature variations away from the actual value (drift) found during checks and/or calibration may show a need for more frequent calibration or the need to replace the device (perhaps with a more durable device). Calibration should be performed at a minimum of two temperatures that bracket the temperature range at which it is used; AND • Perform daily calibration of pH meters against standard buffers; AND • Perform other calibration procedures as necessary to ensure the accuracy of the monitoring instruments; AND • Do finished product sampling and analysis to determine water phase salt, pH, or water activity level, as appropriate, at least once every 3 months (except where such testing is performed as part of monitoring); AND • Review monitoring, corrective action, and verification records within 1 week of preparation to ensure they are complete and

CHAPTER 13: Clostridium botulinum Toxin Formation 287 TABLE 13-4

CONTROL STRATEGY EXAMPLE 4 - PICKLING AND SALTING

This table is an example of a portion of a HACCP plan using “Control Strategy Example 4 - Pickling and Salting.” This example illustrates how a pickled herring processor can control C. botulinum toxin formation. It is provided for illustrative purposes only.

C. botulinum toxin formation may be only one of several significant hazards for this product. Refer to Tables 3-2 and 3-4 (Chapter 3) for other potential hazards (e.g., histamine, environmental and chemical contaminants and pesticides, parasites, and metal fragments).

Example Only See Text for Full Recommendations

(1) (2) (3) (4) (5) (6) (7) (8) (9) (10) CHAPTER 13: ToxinCHAPTER Formation botulinum Clostridium CRITICAL LIMITS MONITORING CRITICAL SIGNIFICANT FOR EACH CORRECTIVE CONTROL RECORDS VERIFICATION HAZARD(S) PREVENTIVE ACTION(S) POINT WHAT HOW FREQUENCY WHO MEASURE Pickling C. botulinum Maximum Finished Collect a Each Quality Continue the Pickling control Daily calibration of toxin formation finished product pH in sample of the pickling control pickling process record the pH meter in the finished product pH in the loin muscle product from tank, each personnel until pH meets Review product the loin muscle each pickling cycle the critical limit monitoring, 288 of 5.0 tank at the corrective action, end of each and verification pickling cycle records within and analyze for 1 week of pH using a pH preparation meter Finished C. botulinum Maximum Cooler air Time and Continuous, Production Adjust or Data logger Check the data product storage toxin cooler temperature temperature with visual employee repair cooler printout logger for accuracy formation temperature: data logger check of and damage and during 40°F recorded Hold and to ensure that finished (based on data once evaluate the it is operational product growth of per day product based before putting storage vegetative on time and into operation; pathogens) temperature of check it daily, at exposure the beginning of operations; and calibrate it once per year

Review monitoring, corrective action, and verification records within 1 week of preparation BIBLIOGRAPHY. • Daniels, R. W. 1991. Applying HACCP to new-generation refrigerated foods at retail We have placed the following references on and beyond. Food Technol. 45:122, 124. display in the Division of Dockets Management, • Dufresne, I., J. P. Smith, J. N. Liu, and I. Tarte. Food and Drug Administration, 5630 Fishers Lane, 2000. Effect of films of different oxygen rm. 1061, Rockville, MD 20852. You may see transmission rate on toxin production by them at that location between 9 a.m. and 4 p.m., Clostridium botulinum type E in vacuum Monday through Friday. As of March 29, 2011, packaged cold and hot smoked trout fillets. J. FDA had verified the Web site address for the Food Saf. 20:251-268. references it makes available as hyperlinks from • Eklund, M. W., G. A. Pelroy, R. Paranjpye, M. the Internet copy of this guidance, but FDA is not E. Peterson, and F. M. Teeny. 1982. Inhibition responsible for any subsequent changes to Non- of Clostridium botulinum types A and E FDA Web site references after March 29, 2011. toxin production by liquid smoke and NaCl in hot-process smoke-flavored fish. J. Food • Association of Food and Drug Officials. 2005. Prot. 44:935-941. Cured, salted, & smoked fish establishments • Essuman, K. M., 1992. Fermented fish in good manufacturing practices, including Africa: a study on processing marketing and Listeria Control Manual. Association of Food consumption. FAO Fisheries technical paper no. and Drug Officials, York, PA. T329. FAO, Rome, Italy. ISBN: 9251032556. 80p. • Baird-Parker, A. C., and B. Freame. 1967. • European Chilled Food Federation. 1997. Combined effect of water activity, pH and Guidelines for good hygienic practice in the temperature on the growth of Clostridium manufacture of chilled foods. Kettering, NN. botulinum from spore and vegetative cell • Farber, J. M. 1991. Microbiological aspects of inocula. J. Appl. Bact. 30:420-429. modified atmosphere packaging technology - • Betts, G. D., and J. E. Getts. 1995. Growth a review. J. Food Prot. 54:58-70. and heat resistance of psychotropic • Garren, D. M., M. A. Harrison, and Y. W. Clostridium botulinum in relation to ‘sous Huang. 1994. Clostridium botulinum type E vide’ products. Food Control 6:57-63. outgrowth and toxin production in vacuum- • Boyd, J. W., and B. A. Southcott. 1971. Effects skin packaged shrimp. Food Microbiol. of sodium chloride on outgrowth and toxin 11:467-472. production of Clostridium botulinum type • Gould, G. W. 1999. Sous vide foods: E in cod homogenates. J. Fish. Res. Bd. conclusions of an ECFF botulinum working Canada. 28:1071-1075. party. Food Control 10:47-51. • Brody, A. L. (ed.). 1989. Controlled/modified • Graham, A. F., D. R. Mason, and M. W. atmosphere/vacuum packaging of foods. Peck. 1996. Predictive model of the effect Food and Nutrition Press, Inc., Trumbull, CT. of temperature, pH and sodium chloride • Crisan, E.V., and A. Sands. 1975. Microflora of on growth from spores of non-proteolytic four fermented fish sauces. Appl. Microbiol. Clostridium botulinum. Int. J. Food Microbiol. 29(1): 106-108. 31:69-85. • Christiansen, L. N., J. Deffner, E. M. Foster, • Hathaway, C.L. 1993. Clostridium botulinum and H. Sugiyama. 1968. Survival and and other Clostridia that produce botulinum. outgrowth of Clostridium botulinum type In Clostridium botulinum Ecology and Control E spores in smoked fish. Appl. Microbiol. in Foods. A.H.W Hauschild and K.L. Dodds 16:133-137. (eds.), Marcel Dekker, New York. 1993.

CHAPTER 13: Clostridium botulinum Toxin Formation 289 • Hauschild, A. H. W., and R. Hilsheimer. 1979. Food Safety and Inspection Service, U.S. Effect of salt content and pH on toxigenesis Department of Agriculture, Washington, DC. by Clostridium botulinum in caviar. J. Food http://www.fsis.usda.gov/OPHS/NACMCF/past/ Prot. 42:245-248. map_ fishery.htm. • Hilderbrand, K. S. 1992. Fish smoking • Peck, M. W. 1997. Clostridium botulinum and procedures for forced convection the safety of refrigerated processed foods smokehouses, Special Report 887. Oregon of extended durability. Trends Food Sci. State Extension Service, Newport, OR. Technol. 8:186-192. • Kautter, D. A., P. K. Lynt, T. Lily, and H. M. • Pelroy, G. A, M. W. Eklund, R. N. Paranjpye, Solomon. 1981. Evaluation of the botulism E. M. Suzuki, and M. E. Peterson. 1982. hazard from nitrogen-packed sandwiches. J. Inhibition of Clostridium botulinum types A Food Prot. 44:59-61. and E toxin formation by sodium nitrite and • Kornacki, J. L., and D. A. Gabis. 1990. sodium chloride in hot process (smoked) Microorganisms and Refrigeration salmon. J. Food Prot. 45:833-841. Temperatures. Dairy, Food & Environ. Sanit. • Peterson, M. E., G. A. Pelroy, R. N. Paranjpye, 10:192-195. F. T. Poysky, J. S. Almond, and M. W. Eklund. • Loha-unchit, K. 1998. How Fish Sauce 1993. Parameters for control of Listeria is Made. Kasma’s Thai Food and Travel. monocytogenes in smoked fishery products: http://www.thaifoodandtravel.com/features/ sodium chloride and packaging method. J. fishsauce1.html. Food Prot. 56:938-943. • Lerke, P., and L. Farber. 1971. Heat • Reddy, N. R., A. Paradis, M. G. Roman, H. M. pasteurization of crab and shrimp from the Solomon, and E. J. Rhodehamel. 1996. Toxin Pacific coast of the United States: public development by Clostridium botulinum in health aspects. J. Food Sci. 36:277-279. modified atmosphere-packaged fresh tilapia fillets during storage. J. Food Sci. 61:632-635. • Lyon, W. J., and C. S. Reddmann. 2000. Bacteria associated with processed • Reddy, N. R., D. J. Armstrong, E. J. crawfish and potential toxin production by Rhodehamel, and D. A. Kautter. 1992. Shelf- Clostridium botulinum type E in vacuum- life extension and safety concerns about packaged and aerobically packaged crawfish fresh fishery products packaged under tails. J. Food Prot. 63:1687-1696. modified atmospheres: a review. J. Food Saf. 12:87-118. • McClure, P. J., M. B. Cole, and J. P. P. M. Smelt. 1994. Effects of water activity and pH • Reddy, N. R., H. M. Solomon, H. Yep, M. G. on growth of Clostridium botulinum. J. Appl. Roman, and E. J. Rhodehamel. 1997. Shelf Bact. Symp. Suppl. 76:105S-114S. life and toxin development by Clostridium botulinum during storage in modified • Moody, M.W., G.J. Flick, R.E. Martin, and A.L. atmosphere-packaged fresh aquacultured Correa. 2000. Smoked, cured, and dried fish. salmon fillets. J. Food Prot. 60:1055-1063. In R.E. Martin, E.P. Carter, G.J. Flick, L.M. Davis, (eds.), Marine & Freshwater Products • Reddy, N. R., M. G. Roman, M. Villanueva, Handbook, 2000. Technomic Publishing Co. H. M. Solomon, D. A. Kautter, and E. J. Lancaster, PA Rhodehamel. 1996. Shelf life and Clostridium botulinum toxin development during storage • National Advisory Committee on of modified atmosphere-packaged fresh Microbiological Criteria for Foods. 1992. catfish fillets. J. Food Sci. 62:878-884. Vacuum or modified atmosphere packaging for refrigerated raw fishery products. • Refrigerated Foods and Microbiological Criteria Committee of the National Food

CHAPTER 13: Clostridium botulinum Toxin Formation 290 Processors Association. 1988. Safety considerations for new generation refrigerated foods. Dairy Food Sanit. 8:5-7. • Rhodehamel, E. J. 1992. FDA’s concerns with sous vide processing. Food Technol. 46:73­ 76.Rhodehamel, E. J., H. M. Solomon, T. Lilly, Jr., D. A. Kautter, and J. T. Peeler. 1991. Incidence and heat resistance of Clostridium botulinum type E spores in menhaden surimi. J. Food Sci. 56:1562-1563, 1592. • Ross, T., and P. Dalgaard. 2004. Secondary Models - A3.1.3. Salt, water-phase salt, and water activity. In R. C. McKellar and L. Xuewen (ed.), Modeling microbial responses in food. CRC Press, Boca Raton, FL. • Schmidt, R. V., R. V. Lechowich, and J. F. Folinazzo. 1961. Growth and toxin production by type E Clostridium botulinum below 40°F. J. Food Sci. 26:626-630. • Segner, W. P, C. F. Schmidt, and J. K. Boltz. 1966. Effect of sodium chloride and pH on the outgrowth of spores of type E Clostridium botulinum at optimal and suboptimal temperatures. Appl. Microbiol.14:49-54. • Skinner, G. E., and J. W. Larkin. 1998. Conservative prediction of time to Clostridium botulinum toxin formation for use with time-temperature indicators to ensure the safety of foods. J. Food Prot. 61:1154-1160. • Sobel, J., et al., 2004.Foodborne botulism in the United States, 1990-2000. Emerging Infectious Diseases. 10(9): 1606-1611. • Sugiyama, H., and K. S. Rutledge. 1978. Failure of Clostridium botulinum to grow in fresh mushrooms packaged in plastic film overwraps with holes. J. Food Prot. 41:348-350. • U.S. Food and Drug Administration. 1996. Import Alert 16-74: automatic detention of salt-cured uneviscerated fish. Department of Health and Human Services, Public Health Service, Food and Drug Administration, Center for Food Safety and Applied Nutrition, College Park, MD.

CHAPTER 13: Clostridium botulinum Toxin Formation 291 NOTES:

CHAPTER 13: Clostridium botulinum Toxin Formation 292