Influence of Several Methodological Factors on the Growth of Clostridium perfringens in Cooling Rate Challenge Studies

2004 ◽  
Vol 67 (6) ◽  
pp. 1128-1132 ◽  
Author(s):  
SARAH SMITH ◽  
VIJAY JUNEJA ◽  
DONALD W. SCHAFFNER
Keyword(s):  
Clostridium Perfringens ◽  
Food Poisoning ◽  
Meat Products ◽  
Ground Beef ◽  
Inoculum Size ◽  
Growth Environment ◽  
Significant Difference ◽  
Plastic Bag ◽  
Major Factors ◽  
Inspection Service

Proper temperature control is essential in preventing Clostridium perfringens food poisoning. The U.S. Department of Agriculture Food Safety and Inspection Service cooling guidelines offer two options for the cooling of meat products: follow a standard time-temperature schedule or validate that alternative cooling regimens result in no more than a 1-log CFU/g increase of C. perfringens and no growth of Clostridium botulinum. The latter option requires laboratory challenge studies to validate the efficacy of a given cooling process. Accordingly, the objective of this study was to investigate the role of several methodological variables that might be encountered during typical C. perfringens challenge studies. Variables studied included plastic bag type (Whirlpak or Spiral Biotech), sealing method (Multivac or FoodSaver), initial spore inoculum size (1 to approximately 3 log CFU/g), and growth environment (ground beef or Trypticase–peptone–glucose–yeast extract [TPGY] broth). The major factors that affected growth were sample bag type and growth environment. Samples incubated in Whirlpak bags showed significantly less growth than those incubated in Spiral Biotech bags, which was likely due to the former bag's greater oxygen permeability. C. perfringens spores showed shorter germination, outgrowth, and lag times and C. perfringens cells showed faster growth rates in ground beef compared with TPGY broth. No significant difference was observed between two different sealing methods. Initial spore inoculum levels in the range studied had no significant effect on final C. perfringens cell concentration.

scholarly journals Evaluation of a Clostridium perfringens Predictive Model, Developed under Isothermal Conditions in Broth, To Predict Growth in Ground Beef during Cooling

2004 ◽  
Vol 70 (5) ◽  
pp. 2728-2733 ◽  
Author(s):  
Sarah Smith ◽  
Donald W. Schaffner
Keyword(s):  
Clostridium Perfringens ◽  
Mathematical Formulation ◽  
Meat Products ◽  
Ground Beef ◽  
Toxin Production ◽  
Performance Standard ◽  
Published Data ◽  
Department Of Agriculture ◽  
Inspection Service ◽  
Good Agreement

ABSTRACT Proper temperature control is essential in minimizing Clostridium perfringens germination, growth, and toxin production. The U.S. Department of Agriculture (USDA) Food Safety and Inspection Service (FSIS) offers two options for the cooling of meat products: follow a standard time-temperature schedule or validate that alternative cooling regimens result in no more than a 1-log10 CFU/g increase of C. perfringens and no growth of Clostridium botulinum. A mathematical model developed by Juneja et al. (Food Microbiol. 16:335-349, 1999) may be helpful in determining if the C. perfringens performance standard has been achieved, but this model has not been extensively validated. The objective of this study was to validate the Juneja 1999 model in ground beef under a variety of changing temperature and temperature abuse situations. The Juneja 1999 model consistently underpredicted growth of C. perfringens during exponential cooling of ground beef. The model also underpredicted growth of C. perfringens in ground beef cooled at two different rates. The results presented here show generally good agreement with published data on the growth of C. perfringens in similar products. The model error may be due to faster-than-expected exponential growth rates in ground beef during cooling or an error in the mathematical formulation of the model.

scholarly journals Incidence and contamination level of Clostridium perfringens in meat and meat products sold in Sakarya province of Turkey

2021 ◽  
Vol 7 (3) ◽  
pp. 172-178
Author(s):  
Serap Coşansu ◽  
Şeyma Şeniz Ersöz
Keyword(s):  
Clostridium Perfringens ◽  
Meat Product ◽  
Meat Products ◽  
Ground Beef ◽  
Chicken Meat ◽  
Contamination Level ◽  
Biochemical Tests ◽  
Contamination Levels ◽  
Emulsified Meat ◽  
Cooked Meat

Totally 101 meat and meat product samples obtained from local markets and restaurants were analyzed for incidence and contamination level of Clostridium perfringens. The typical colonies grown anaerobically on Tryptose Sulfite Cycloserine Agar supplemented with 4-Methyliumbelliferyl (MUP) were confirmed by biochemical tests. Forty-eight of the samples (47.5%) were contaminated with C. perfringens. The highest incidence of the pathogen was determined in uncooked meatball samples (72.2%) followed by ground beef samples (61.3%). The incidence of C. perfringens in chicken meat, cooked meat döner, cooked chicken döner and emulsified meat product samples were 33.3, 33.3, 28.6 and 16.7%, respectively. Thirteen out of 101 samples (12.9%) yielded typical colonies on TSC-MUP Agar, but could not be confirmed as C. perfringens. Average contamination levels in sample groups ranged from 8.3 to 1.5×102 cfu/g, with the highest ground beef and the lowest chicken meat.

Growth and Sporulation Potential of Clostridium perfringens in Aerobic and Vacuum-Packaged Cooked Beef

1994 ◽  
Vol 57 (5) ◽  
pp. 393-398 ◽  
Author(s):  
V. K. JUNEJA ◽  
B. S. MARMER ◽  
A. J. MILLER
Keyword(s):  
Clostridium Perfringens ◽  
Food Poisoning ◽  
Ground Beef ◽  
Viable Count ◽  
Anaerobic Conditions ◽  
Vegetative Cells ◽  
Aerobic Conditions ◽  
C Storage ◽  
Meat Samples ◽  
Aerobic And Anaerobic

Growth of Clostridium perfringens in aerobic-and anaerobic-(vacuum) packaged cooked ground beef was investigated. Autoclaved ground beef was inoculated with ~3.0-log10 CFU/g of C. perfringens, packaged and stored at various temperatures. Vegetative cells and heat-resistant spores were enumerated by plating unheated and heated (75°C for 20 min) meat samples on tryptose-sulfite-cycloserine agar. Clostridium perfringens grew to >7 logs within 12 h at 28, 37 and 42°C under anaerobic atmosphere and at 37 and 42°C under aerobic conditions. At 28°C under aerobic conditions, growth was relatively slow and total viable count increased to >6 logs within 36 h. Similarly, growth at 15°C in air was both slower and less than under vacuum. Regardless of packaging, the organism either declined or did not grow at 4, 8 and 12°C. Spores were not found at <12°C. Spores were detected as early as 8 h at 42°C under anaerobic conditions, but in general, the type of atmosphere had little influence on sporulation at ≥28°C. Temperature abuse (28°C storage) of refrigerated products for 6 h will not permit C. perfringens growth. However, cyclic and static temperature abuse of such products for relatively long periods may lead to high and dangerous numbers of organisms. Reheating such products to an internal temperature of 65°C before consumption would prevent food poisoning since the vegetative cells were killed.

Inhibition of Clostridium perfringens Growth by Potassium Lactate during an Extended Cooling of Cooked Uncured Ground Turkey Breasts

2013 ◽  
Vol 76 (11) ◽  
pp. 1972-1976 ◽  
Author(s):  
KATHERINE M. KENNEDY ◽  
ANDREW L. MILKOWSKI ◽  
KATHLEEN A. GLASS
Keyword(s):  
Clostridium Perfringens ◽  
Clostridium Botulinum ◽  
Meat Products ◽  
Sodium Lactate ◽  
Limited Growth ◽  
Treatment Groups ◽  
Potassium Lactate ◽  
Inspection Service ◽  
Spore Forming Bacteria ◽  
The U.S

The U.S. Department of Agriculture's Food Safety and Inspection Service compliance guideline known as Appendix B specifies chilling time and temperature limits for cured and uncured meat products to inhibit growth of spore-forming bacteria, particularly Clostridium perfringens. Sodium lactate and potassium lactate inhibit toxigenic growth of Clostridium botulinum, and inhibition of C. perfringens has been reported. In this study, a cocktail of spores of three C. perfringens strains (ATCC 13124, ATCC 12915, and ATCC 12916) were inoculated into 100-g samples of ground skinless, boneless turkey breast formulated to represent deli-style turkey breast. Three treatment groups were supplemented with 0 (control), 1, or 2% potassium lactate (pure basis), cooked to 71°C, and assayed for C. perfringens growth during 10 or 12 h of linear cooling to 4°C. In control samples, populations of C. perfringens increased 3.8 to 4.7 log CFU/g during the two chilling protocols. The 1% potassium lactate treatment supported only a 2.5- to 2.7-log increase, and the 2% potassium lactate treatment limited growth to a 0.56- to 0.70-log increase. When compared with the control, 2% potassium lactate retarded growth by 2.65 and 4.21 log CFU/g for the 10- and 12-h cooling protocols, respectively. These results confirm that the addition of 2% potassium lactate inhibits growth of C. perfringens and that potassium lactate can be used as an alternative to sodium nitrite for safe extended cooling of uncured meats.

scholarly journals Roles of DacB and Spm Proteins in Clostridium perfringens Spore Resistance to Moist Heat, Chemicals, and UV Radiation

2008 ◽  
Vol 74 (12) ◽  
pp. 3730-3738 ◽  
Author(s):  
Daniel Paredes-Sabja ◽  
Nahid Sarker ◽  
Barbara Setlow ◽  
Peter Setlow ◽  
Mahfuzur R. Sarker
Keyword(s):  
Water Content ◽  
Uv Radiation ◽  
Clostridium Perfringens ◽  
Nitrous Acid ◽  
Food Poisoning ◽  
Cross Linking ◽  
Gene Products ◽  
Moist Heat ◽  
Spore Resistance ◽  
Major Factors

ABSTRACT Clostridium perfringens food poisoning is caused mainly by enterotoxigenic type A isolates that typically possess high spore heat resistance. Previous studies have shown that α/β-type small, acid-soluble proteins (SASP) play a major role in the resistance of Bacillus subtilis and C. perfringens spores to moist heat, UV radiation, and some chemicals. Additional major factors in B. subtilis spore resistance are the spore's core water content and cortex peptidoglycan (PG) structure, with the latter properties modulated by the spm and dacB gene products and the sporulation temperature. In the current work, we have shown that the spm and dacB genes are expressed only during C. perfringens sporulation and have examined the effects of spm and dacB mutations and sporulation temperature on spore core water content and spore resistance to moist heat, UV radiation, and a number of chemicals. The results of these analyses indicate that for C. perfringens SM101 (i) core water content and, probably, cortex PG structure have little if any role in spore resistance to UV and formaldehyde, presumably because these spores’ DNA is saturated with α/β-type SASP; (ii) spore resistance to moist heat and nitrous acid is determined to a large extent by core water content and, probably, cortex structure; (iii) core water content and cortex PG cross-linking play little or no role in spore resistance to hydrogen peroxide; (iv) spore core water content decreases with higher sporulation temperatures, resulting in spores that are more resistant to moist heat; and (v) factors in addition to SpmAB, DacB, and sporulation temperature play roles in determining spore core water content and thus, spore resistance to moist heat.

Effect of Refrigerating Delayed Shipments of Raw Ground Beef on the Detection of Salmonella Typhimurium†

2005 ◽  
Vol 68 (8) ◽  
pp. 1581-1586 ◽  
Author(s):  
NEELAM NARANG ◽  
MARK L. TAMPLIN ◽  
WILLIAM C. CRAY
Keyword(s):  
Salmonella Typhimurium ◽  
Ground Beef ◽  
Data Logger ◽  
Eastern Regional Research ◽  
Significant Difference ◽  
Microbiological Laboratory ◽  
Shipping Containers ◽  
Inspection Service ◽  
The U.S ◽  
Ice Pack

In eight separate trials, four groups of raw ground beef samples were inoculated with 0.04 to 0.3 CFU/g of Salmonella Typhimurium (DT 104). Each group consisted of four 25-g samples (three inoculated and one uninoculated). After inoculation, these samples were shipped by overnight courier in shipping containers with ice packs from the U.S. Department of Agriculture (USDA), Eastern Regional Research Center, in Wyndmoor, Pa., to the U.S. Food Safety and Inspection Service (FSIS), Eastern Laboratory, in Athens, Ga. A total of 128 samples (32 in each of four groups) were shipped. A temperature data logger was placed inside each shipping container to record the temperature during shipping and storage. The first group of ground beef samples was analyzed within approximately 1 h of arrival. The second group of samples was left in the original containers, with a gel ice pack, for 24 h before processing. The third and fourth groups of samples were removed from the original shipping containers and stored at room temperature (21 ± 2°C) for 6 h and then in a refrigerator at 4 ± 2°C for 24 and 48 h, respectively, before analysis. The samples were analyzed for the presence of Salmonella according to the USDA/FSIS Microbiological Laboratory Guidebook, chapter 4.02. There was no significant difference in the presence and levels of Salmonella in ground beef among the four test groups. These data show that it is acceptable to process the late-arriving ground beef samples for the detection of Salmonella if they are kept in a refrigerator (4 ± 2°C) for 24 to 48 h or when the shipments arrive late (24 h in the container with ice pack).

scholarly journals Comparative Experiments To Examine the Effects of Heating on Vegetative Cells and Spores of Clostridium perfringens Isolates Carrying Plasmid Genes versus Chromosomal Enterotoxin Genes

2000 ◽  
Vol 66 (8) ◽  
pp. 3234-3240 ◽  
Author(s):  
Mahfuzur R. Sarker ◽  
Robert P. Shivers ◽  
Shauna G. Sparks ◽  
Vijay K. Juneja ◽  
Bruce A. McClane
Keyword(s):  
Clostridium Perfringens ◽  
Gastrointestinal Disease ◽  
Food Poisoning ◽  
Meat Products ◽  
Gastrointestinal Diseases ◽  
Vegetative Cells ◽  
Food Borne ◽  
Enterotoxin Genes ◽  
Plasmid Genes ◽  
High Degree

ABSTRACT Clostridium perfringens enterotoxin (CPE) is an important virulence factor for both C. perfringens type A food poisoning and several non-food-borne human gastrointestinal diseases. Recent studies have indicated that C. perfringensisolates associated with food poisoning carry a chromosomalcpe gene, while non-food-borne human gastrointestinal disease isolates carry a plasmid cpe gene. However, no explanation has been provided for the strong associations between certain cpe genotypes and particular CPE-associated diseases. Since C. perfringens food poisoning usually involves cooked meat products, we hypothesized that chromosomalcpe isolates are so strongly associated with food poisoning because (i) they are more heat resistant than plasmid cpeisolates, (ii) heating induces loss of the cpe plasmid, or (iii) heating induces migration of the plasmid cpe gene to the chromosome. When we tested these hypotheses, vegetative cells of chromosomal cpe isolates were found to exhibit, on average approximately twofold-higher decimal reduction values (Dvalues) at 55°C than vegetative cells of plasmid cpeisolates exhibited. Furthermore, the spores of chromosomalcpe isolates had, on average, approximately 60-fold-higherD values at 100°C than the spores of plasmidcpe isolates had. Southern hybridization and CPE Western blot analyses demonstrated that all survivors of heating retained theircpe gene in its original plasmid or chromosomal location and could still express CPE. These results suggest that chromosomalcpe isolates are strongly associated with food poisoning, at least in part, because their cells and spores possess a high degree of heat resistance, which should enhance their survival in incompletely cooked or inadequately warmed foods.

scholarly journals Visual Detection of Clostridium perfringens Alpha Toxin by Combining Nanometer Microspheres with Smart Phones

2020 ◽  
Vol 8 (12) ◽  
pp. 1865
Author(s):  
Aiping Cao ◽  
Heng Chi ◽  
Jingxuan Shi ◽  
Ruiqi Sun ◽  
Kang Du ◽  
...  
Keyword(s):  
Clostridium Perfringens ◽  
Detection Method ◽  
Detection System ◽  
Food Poisoning ◽  
Meat Products ◽  
Nano Silica ◽  
Silica Microspheres ◽  
A Cell ◽  
Important Virulence Factor ◽  
Hemorrhagic Enteritis

Clostridium perfringens α toxin (CPA) is an important virulence factor that causes livestock hemorrhagic enteritis and food poisoning by contaminated meat products. In this study, the nano-silica microspheres combined with smartphone image processing technology was developed to realize real-time CPA detection. First, the N-terminal and C-terminal domain of the CPA toxin (CPAC3 and CPAN) and their anti-sera were prepared. The silica microspheres coupled with the antibody of CPAC3 was prepared to capture the toxin that existed in the detection sample and the fluorescent-labeled antibody of CPAN was incubated. Moreover, the fluorescent pictures of gray value were performed in a cell phone app, corresponding to toxin concentration. The new assay takes 90 min to perform and can detect CPA as little as 32.8 ng/mL. Our results showed a sensitive, stable, and convenient CPA detection system, which provides a novel detection method of native CPA in foods.

Effects of Nitrite and Erythorbate on Clostridium perfringens Growth during Extended Cooling of Cured Ham

2017 ◽  
Vol 80 (10) ◽  
pp. 1697-1704 ◽  
Author(s):  
Katie J. Osterbauer ◽  
Amanda M King ◽  
Dennis L Seman ◽  
Andrew L. Milkowski ◽  
Kathleen A. Glass ◽  
...  
Keyword(s):  
Clostridium Perfringens ◽  
Phase 1 ◽  
Growth Control ◽  
Meat Products ◽  
Phase 2 ◽  
Department Of Agriculture ◽  
Cooling Period ◽  
Poultry Products ◽  
Heat Treated ◽  
Inspection Service

ABSTRACT To control the growth of Clostridium perfringens in cured meat products, the meat and poultry industries commonly follow stabilization parameters outlined in Appendix B, “Compliance Guidelines for Cooling Heat-Treated Meat and Poultry Products (Stabilization)” (U.S. Department of Agriculture, Food Safety and Inspection Service [USDA-FSIS], 1999) to achieve cooling (54.4 to 4.4°C) within 15 h after cooking. In this study, extended cooling times and their impact on C. perfringens growth were examined. Phase 1 experiments consisted of cured ham with 200 mg/kg ingoing sodium nitrite and 547 mg/kg sodium erythorbate following five bilinear cooling profiles: a control (following Appendix B guidelines: stage A cooling [54.4 to 26.7°C] for 5 h, stage B cooling [26.7 to 4.4°C] for 10 h), extended stage A cooling for 7.5 or 10 h, and extended stage B cooling for 12.5 or 15 h. A positive growth control with 0 mg/kg nitrite added (uncured) was also included. No growth was observed in any treatment samples except the uncured control (4.31-log increase within 5 h; stage A). Phase 2 and 3 experiments were designed to investigate the effects of various nitrite and erythorbate concentrations and followed a 10-h stage A and 15-h stage B bilinear cooling profile. Phase 2 examined the effects of nitrite concentrations of 0, 50, 75, 100, 150, and 200 mg/kg at a constant concentration of erythorbate (547 mg/kg). Results revealed changes in C. perfringens populations for each treatment of 6.75, 3.59, 2.43, −0.38, −0.48, and −0.50 log CFU/g, respectively. Phase 3 examined the effects of various nitrite and erythorbate concentrations at 100 mg/kg nitrite with 0 mg/kg erythorbate, 100 with 250, 100 with 375, 100 with 547, 150 with 250, and 200 with 250, respectively. The changes in C. perfringens populations for each treatment were 4.99, 2.87, 2.50, 1.47, 0.89, and −0.60 log CFU/g, respectively. Variability in C. perfringens growth for the 100 mg/kg nitrite with 547 mg/kg erythorbate treatment was observed between phases 2 and 3 and may have been due to variations in treatment pH and NaCl concentrations. This study revealed the importance of nitrite and erythorbate for preventing growth of C. perfringens during a much longer (25 h) cooling period than currently specified in the USDA-FSIS Appendix B.

Growth Potential of Clostridium perfringens from Spores in Acidified Beef, Pork, and Poultry Products during Chilling†

2013 ◽  
Vol 76 (1) ◽  
pp. 65-71 ◽  
Author(s):  
VIJAY K. JUNEJA ◽  
DAVID A. BAKER ◽  
H. THIPPAREDDI ◽  
O. PETER SNYDER ◽  
TIM B. MOHR
Keyword(s):  
Clostridium Perfringens ◽  
Ground Beef ◽  
Growth Potential ◽  
Department Of Agriculture ◽  
Inoculum Level ◽  
Isothermal Temperature ◽  
Ph Values ◽  
Poultry Products ◽  
Challenge Tests ◽  
Inspection Service

The ability of Clostridium perfringens to germinate and grow in acidified ground beef as well as in 10 commercially prepared acidified beef, pork, and poultry products was assessed. The pH of ground beef was adjusted with organic vinegar to achieve various pH values between 5.0 and 5.6; the pH of the commercial products ranged from 4.74 to 6.35. Products were inoculated with a three-strain cocktail of C. perfringens spores to achieve ca. 2-log (low) or 4-log (high) inoculum levels, vacuum packaged, and cooled exponentially from 54.4 to 7.2°C for 6, 9, 12, 15, 18, or 21 h to simulate abusive cooling; the U.S. Department of Agriculture, Food Safety and Inspection Service (USDA-FSIS) recommends a cooling time of 6.5 h. Total germinated C. perfringens populations were determined after plating on tryptose-sulfite-cycloserine agar and incubating the plates anaerobically at 37°C for 48 h. In addition, C. perfringens growth from spores was assessed at an isothermal temperature of 44°C. Growth from spores was inhibited in ground beef with a pH of 5.5 or below, even during extended cooling from 54.4 to 7.2°C in 21 h. In ground beef with a pH of 5.6, the growth was >1 log after 18 h of cooling from 54.4 to 7.2°C. However, 15 h of cooling controlled the growth to <1 log, regardless of the inoculum level. In addition, no growth was observed in any product with a pH ranging from 4.74 to 5.17, both during exponential abusive cooling periods of up to 21 h and during storage for 21 h at 44°C. While <1-log growth of C. perfringens from spores was observed in the pH 5.63 product cooled exponentially from 54.4 to 7.2°C in 15 h or less, the pH 6.35 product supported growth, even after 6 h of cooling from 54.4 to 7.2°C. These challenge tests demonstrate that adjustment of ground beef to pH of 5.5 or less and of barbeque products to pH of 5.63 or less inhibits C. perfringens spore germination and outgrowth during extended cooling periods from 54.4 to 7.2°C up to 15 h. Therefore, safe cooling periods for products with homogeneous, lower pHs can be substantially longer.

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