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Chapter 13: Clostridium perfringens

Updated: 07/18/07
Potential Food Safety Hazard Contents
Food poisoning caused by Clostridium perfringens may occur when foods such as meat or poultry are cooked and held without maintaining adequate heating or refrigeration before serving. The presence of small numbers of C. perfringens is not uncommon in raw meats, poultry, dehydrated soups and sauces, raw vegetables, and spices. Because the spores of some strains are resistant to temperatures as high as 100ēC for more than l h, their presence in foods may be unavoidable. Furthermore, the oxygen level may be sufficiently reduced during cooking to permit growth of the clostridia. Spores that survive cooking may germinate and grow rapidly in foods that are inadequately refrigerated after cooking. Thus, when clinical and epidemiological evidence suggests that C. perfringens is the cause of a food poisoning outbreak, the presence of hundreds of thousands or more of these organisms per gram of food substantiates the diagnosis.

Illness typically occurs 8-15 h after ingestion of the contaminated food. The symptoms, which include intense abdominal cramps, gas, and diarrhea (nausea and vomiting are rare), have been attributed to a protein enterotoxin produced during sporulation of the organism in the intestine. The enterotoxin can be detected in sporulating cultures, and a method for this purpose is included. A high correlation has been established between the ability of C. perfringens strains to produce enterotoxin and their ability to cause food poisoning. However, it is difficult to obtain consistent sporulation with some strains (Rhodehamel and Harmon, 1998).

Control Measures Contents
Control measures emphasize proper food preparation and storage techniques, especially temperature control. Control measures include:
  1. Rapid, uniform cooling of cooked foods to <10ºC (50ºF) within 2-3 h;
  2. Hot holding of cooked foods at or above 60ºC (140ºF);
  3. Reheating cooled or chilled foods to a minimum internal temperature of 75ºC (167ºF) immediately before serving;
  4. Not leaving foods at room temperature or thawing frozen foods at room temperature;
  5. Preventing cross-contamination of cooked foods with bacteria from raw foods by using separate food-contact surfaces for preparing raw and cooked food items, or by thoroughly cleaning and sanitizing food-contact surfaces after being used for raw products;
  6. Maintaining food preparation areas so that they are free of soil and dust;
  7. Cleaning and sanitizing meat slicers, meat-cutting equipment, food-contact surfaces, and other equipment after use; and
  8. Using good personal hygiene methods, and thoroughly washing hands frequently when handling food products, especially after handling raw products and before handling cooked products (Labbe, 1989; Reed, 1994).
FDA Guideline Contents
FDA to assess situations on a case by case basis.

Growth Contents
Table A-4. Limiting conditions for pathogen growth.

Heat Resistance Contents
Heat resistance of C. perfringens spores.
Temp.
D-Value
Strain
 Medium
Reference
(ºC)
(ºF)
(min.)
55
131
179
NCTC 8798
Ground beef
 Roy et al., 1981
57
134.6
34.9
NCTC 8798
Ground beef
 Roy et al., 1981
59
138.2
16.9
NCTC 8238
Ground beef
 Roy et al., 1981
61
141.8
3.9
NCTC 8238
Ground beef
 Roy et al., 1981
98.9
210.0
31.4
NCTC 8798
Beef gravy

Bradshaw et al., 1977
100
212
17.6
NCTC 8238
SEC broth
 Weiss and Strong, 1967
104.4
219.9
8.0
NCTC 10240
Beef gravy

Bradshaw et al., 1977
110.0
230
0.95
NCTC 10240
Beef gravy
 Bradshaw et al., 1977
115.6
240.1
0.21
NCTC 10240
Beef gravy
 Bradshaw et al., 1977

Analytical Procedures Contents
Food sampling and preparation of sample homogenate (USFDA) Contents
Definition of Terms (HC Appendix A); Collection of samples (HC Appendix B); Supplement to All Methods in the HC Compendium: General Microbiological Guidance (HC Appendix I)  General Microbiological guidance on Pre-warming of Broths in All Qualitative Methods in the [HC] Compendium (HC Supplement to Appendix I) Contents
Clostridium perfringens (USFDA) Contents
Enumeration of Clostridium perfringens in foods (HC MFHPB-23)  Contents
Other analytical procedures Contents
  • Clostridium perfringens from shellfish (AOAC, 1995c)
  • Clostridium perfringens in foods: alpha-toxin estimation method (AOAC, 1995b)
  • Clostridium perfringens in foods: Microbiological method (AOAC, 1995a)
  • MPN method for C. perfringens in shellfish (Abeyta and Wetherington, 1994)

Commercial Test Products Contents
Commercial test products for C. perfringens.

Test Kit

Analytical Technique

Approx. Total Test Time1

Supplier
Clostridium Perfringens Test
[A presumptive test for Clostridium perfringens]		 Uses prepared traditional media 48 h Biomedix
Contact: Claver Bundac
1105 #F North Golden Springs Dr.
Diamond Bar, CA 91765
Phone: 800/674-8648 #4282; 909/396-0244  
E-mail: cb4biomedx@aol.com

ISO-GRID Method for Clostridium perfringens Count using Modified TSC agar

Membrane filtration with selective culture medium

24-72 h (24 h for presumptive enumeration and 48 h additional to confirm presumptive positive results)

Neogen Corporation 
620 Lesher Pl. 
Lansing, MI 48912 
Phone: 517/372-9200 
E-mail: NeogenCorp@aol.com 
Web: www.neogen.com/isogridgen.htm

PET-RPLA TD930 
[Used to identify C. perfringens type A enterotoxin]

Reversed passive latex agglutination

24 h (feces)
48 h (bacterial culture)

Oxoid, Inc. 
Contact: Jim Bell 
217 Colonnade Rd. 
Nepean, Ontario K2E 7K3 
Canada 
Phone: 613/226-1318 
E-mail: jbell@oxoid.ca
1Includes enrichment

References Contents
Abeyta, C. and Wetherington, J. 1994. Iron milk medium for recovering Clostridium perfringens from shellfish: Collaborative study. JAOAC 77:351-356.

Andrews, W.H., and June, G.A. 1998. Food sampling and preparation of sample homogenate, Ch. 1. In Food and Drug Administration Bacteriological Analytical Manual, 8th ed. (revision A), (CD-ROM version). R.L. Merker (Ed.). AOAC International, Gaithersburg, MD.

AOAC. 1995a . Clostridium perfringens in foods: microbiological method. Sec. 17.7.02, Method 976.30. In Official Methods of Analysis of AOAC International, 16th ed., P.A. Cunniff (Ed.), p. 48-50. AOAC International, Gaithersburg, MD.

AOAC. 1995b . Clostridium perfringens in foods: Alpha toxin estimation method. Sec. 17.7.03, Method 974.38. In Official Methods of Analysis of AOAC International, 16th ed., P.A. Cunniff (Ed.), p. 50-51. AOAC International, Gaithersburg, MD.

AOAC. 1995c . Clostridium Perfringens from shellfish: Iron milk method. Sec. 17.7.04, Method 993.10. In Official Methods of Analysis of AOAC International, 16th ed., P.A. Cunniff (Ed.), p. 51-52. AOAC International, Gaithersburg, MD.

Bradshaw, J.G., Peeler, J.T., and Twedt, R.M. 1977. Thermal inactivation of ileal loop-reactive Clostridium perfringens type A strains in phosphate buffer and beef gravy. Appl. Environ. Microbiol. 34(3):280-284.

Labbe, R. 1989. Clostridium perfringens. Ch. 5, In Foodborne Bacterial Pathogens, M.P. Doyle (Ed.), p. 191-234. Marcel Dekker, Inc., New York.

Merker, R.L. (Ed.). 1998. Media and Reagents, Appendix 3. In Food and Drug Administration Bacteriological Analytical Manual, 8th ed. (revision A), (CD-ROM version). AOAC International, Gaithersburg, MD.

Reed, G.H. 1994. Foodborne illness (Part 3): Clostridium perfringens gastroenteritis. Dairy, Food and Environmental San. 14(1):16-17.

Rhodehamel, E.J. and Harmon, S.M. 1998. Clostridium perfringens. Ch. 16. In Food and Drug Administration Bacteriological Analytical Manual, 8th ed. (revision A), (CD-ROM version). R.L. Merker (Ed.). AOAC International, Gaithersburg, MD.

Roy, R.J., Busta, F.F., and Thompson, D.R. 1981. Thermal inactivation of Clostridium botulinum after growth at several constant and linearly rising temperatures. J. Food Sci. 46(5):1586-1591.

Weiss, K.F. and Strong, D.H. 1967. Some properties of heat-resistant strains of Clostridium perfringens. I. Heat resistance and toxigenicity. J. Bacteriol. 93(1):21-26.

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Updated: 07/18/07

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Pamela D. Tom, SeafoodNIC Director
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