Inhibition of Germination and Outgrowth of Clostridium perfringens Spores by Lactic Acid Salts during Cooling of Injected Turkey†

2007 ◽  
Vol 70 (4) ◽  
pp. 923-929 ◽  
Author(s):  
PADMANABHA REDDY VELUGOTI ◽  
LALIT K. BOHRA ◽  
VIJAY K. JUNEJA ◽  
HARSHAVARDHAN THIPPAREDDI
Keyword(s):  
Lactic Acid ◽  
Clostridium Perfringens ◽  
Spore Germination ◽  
Calcium Lactate ◽  
Heat Treated ◽  
Potassium Lactate ◽  
Additional Protection ◽  
Spore Population ◽  
Sodium And Potassium ◽  
Acid Salts

Inhibition of Clostridium perfringens spore germination and outgrowth by lactic acid salts (calcium, potassium, and sodium) during exponential cooling of injected turkey product was evaluated. Injected turkey samples containing calcium lactate, potassium lactate, or sodium lactate (1.0, 2.0, 3.0, or 4.8% [w/w]), along with a control (product without lactate), were inoculated with a three-strain cocktail of C. perfringens spores to achieve a final spore population of 2.5 to 3.0 log CFU/g. The inoculated product was heat treated and exponentially cooled from 54.5 to 7.2°C within 21, 18, 15, 12, 9, or 6.5 h. Cooling of injected turkey (containing no antimicrobials) resulted in C. perfringens germination and an outgrowth of 0.5, 2.4, 3.4, 5.1, 5.8, and 5.8 log CFU/g when exponentially cooled from 54.4 to 7.2°C in 6.5, 12, 15, 18, and 21 h, respectively. The incorporation of antimicrobials (lactates), regardless of the type (Ca, Na, or K salts), inhibited the germination and outgrowth of C. perfringens spores at all the concentrations evaluated (1.0, 2.0, 3.0, and 4.8%) compared to the injected turkey without acetate (control). Increasing the concentrations of the antimicrobials resulted in a greater inhibition of the spore germination and outgrowth in the products. In general, calcium lactate was more effective in inhibiting the germination and outgrowth of C. perfringens spores at ≥1.0% concentration than were sodium and potassium lactates. Incorporation of these antimicrobials in cooked, ready-to-eat turkey products can provide additional protection in controlling the germination and outgrowth of C. perfringens spores during cooling (stabilization).

Evaluation of the Microbial Quality of Tajik Sambusa and Control of Clostridium perfringens Germination and Outgrowth by Buffered Sodium Citrate and Potassium Lactate†

2008 ◽  
Vol 71 (1) ◽  
pp. 77-82 ◽  
Author(s):  
SHAKHLO N. YARBAEVA ◽  
PADMANABHA R. VELUGOTI ◽  
HARSHAVARDHAN THIPPAREDDI ◽  
JULIE A. ALBRECHT
Keyword(s):  
Organic Acid ◽  
Clostridium Perfringens ◽  
Spore Germination ◽  
Sodium Citrate ◽  
Ground Beef ◽  
E Coli ◽  
Potassium Lactate ◽  
Before And After ◽  
Organic Acid Salts ◽  
Acid Salts

Clostridium perfringens spore destruction, aerobic plate counts (APCs), and counts of Enterobacteriaceae, coliforms, and Escherichia coli during baking of sambusa (a traditional Tajik food) were evaluated. Control of germination and outgrowth of C. perfringens spores in sambusa during cooling at room or refrigerated temperatures was evaluated using organic acid salts (buffered sodium citrate [Ional] and 1 and 2% potassium lactate, wt/wt). Sambusa were prepared with 40 g of either inoculated or noninoculated meat and baked for 45 min at 180°C. For evaluation of destruction of C. perfringens spores during heating and germination and outgrowth of spores during cooling, ground beef was inoculated and mixed with a three-strain cocktail of C. perfringens spores. Aerobic bacteria, Enterobacteriaceae, coliforms, and E. coli were enumerated in noninoculated sambusa before and after baking and after cooling at room or refrigeration temperatures. After baking, APCs and Enterobacteriaceae and coliform counts were reduced by 4.32, 2.55, and 1.96 log CFU/g, respectively. E. coli counts were below detectable levels in ground beef and sambusa samples. Enterobacteriaceae, coliform, and E. coli counts were below detectable levels (<0.04 log CFU/g) in sambusa after cooling by both methods. Total C. perfringens populations increased (4.67 log CFU/g) during cooling at room temperature, but minimal increases (0.31 log CFU/g) were observed during cooling under refrigeration. Incorporation of 2% (wt/wt) buffered sodium citrate controlled C. perfringens spore germination and outgrowth (0.25 log CFU/g), whereas incorporation of up to 2% (wt/wt) potassium lactate did not prevent C. perfringens spore germination and outgrowth. Incorporation of organic acid salts at appropriate concentrations can prevent germination and outgrowth of C. perfringens in improperly cooled sambusa.

Predictive Model for Clostridium perfringens Growth in Roast Beef during Cooling and Inhibition of Spore Germination and Outgrowth by Organic Acid Salts†‡

2005 ◽  
Vol 68 (12) ◽  
pp. 2594-2605 ◽  
Author(s):  
MARCOS X. SÁNCHEZ-PLATA ◽  
ALEJANDRO AMÉZQUITA ◽  
ERIN BLANKENSHIP ◽  
DENNIS E. BURSON ◽  
VIJAY JUNEJA ◽  
...  
Keyword(s):  
Organic Acid ◽  
Predictive Model ◽  
Clostridium Perfringens ◽  
Spore Germination ◽  
Sodium Citrate ◽  
Sodium Lactate ◽  
Processed Meat ◽  
Roast Beef ◽  
Organic Acid Salts ◽  
Acid Salts

Spores of foodborne pathogens can survive traditional thermal processing schedules used in the manufacturing of processed meat products. Heat-activated spores can germinate and grow to hazardous levels when these products are improperly chilled. Germination and outgrowth of Clostridium perfringens spores in roast beef during chilling was studied following simulated cooling schedules normally used in the processed-meat industry. Inhibitory effects of organic acid salts on germination and outgrowth of C. perfringens spores during chilling and the survival of vegetative cells and spores under abusive refrigerated storage was also evaluated. Beef top rounds were formulated to contain a marinade (finished product concentrations: 1% salt, 0.2% potassium tetrapyrophosphate, and 0.2% starch) and then ground and mixed with antimicrobials (sodium lactate and sodium lactate plus 2.5% sodium diacetate and buffered sodium citrate and buffered sodium citrate plus 1.3% sodium diacetate). The ground product was inoculated with a three-strain cocktail of C. perfringens spores (NCTC 8238, NCTC 8239, and ATCC 10388), mixed, vacuum packaged, heat shocked for 20 min at 75°C, and chilled exponentially from 54.5 to 7.2°C in 9, 12, 15, 18, or 21 h. C. perfringens populations (total and spore) were enumerated after heat shock, during chilling, and during storage for up to 60 days at 10°C using tryptose-sulfite-cycloserine agar. C. perfringens spores were able to germinate and grow in roast beef (control, without any antimicrobials) from an initial population of ca. 3.1 log CFU/g by 2.00, 3.44, 4.04, 4.86, and 5.72 log CFU/g after 9, 12, 15, 18, and 21 h of exponential chilling. A predictive model was developed to describe sigmoidal C. perfringens growth curves during cooling of roast beef from 54.5 to 7.2°C within 9, 12, 15, 18, and 21 h. Addition of antimicrobials prevented germination and outgrowth of C. perfringens regardless of the chill times. C. perfringens spores could be recovered from samples containing organic acid salts that were stored up to 60 days at 10°C. Extension of chilling time to ≥9 h resulted in >1 log CFU/g growth of C. perfringens under anaerobic conditions in roast beef. Organic acid salts inhibited outgrowth of C. perfringens spores during chilling of roast beef when extended chill rates were followed. Although C. perfringens spore germination is inhibited by the antimicrobials, this inhibition may represent a hazard when such products are incorporated into new products, such as soups and chili, that do not contain these antimicrobials, thus allowing spore germination and outgrowth under conditions of temperature abuse.

Growth of Heat-Treated Enterotoxin-PositiveClostridium perfringens and the Implications for Safe Cooling Rates

2004 ◽  
Vol 67 (1) ◽  
pp. 83-89 ◽  
Author(s):  
KARIN G. ANDERSEN ◽  
TINA B. HANSEN ◽  
SUSANNE KNØCHEL
Keyword(s):  
Clostridium Perfringens ◽  
Spore Germination ◽  
Generation Time ◽  
Growth Characteristics ◽  
Heat Inactivation ◽  
Lag Phase ◽  
Temperature Growth ◽  
Generation Times ◽  
Heat Treated ◽  
Higher Temperature

Clostridium perfringens 790-94 and 44071.C05 carrying a chromosomal and a plasmid cpe gene, respectively, were used to determine differences in heat resistance and growth characteristics between the genotypes. Heat inactivation experiments were conducted using an immersed coil apparatus. Spore germination, outgrowth, and lag phase, together named GOL time, as well as generation times were determined during constant temperatures in fluid thioglycollate (FTG) medium as well as in vacuum-packed, heat-treated minced turkey. GOL time and growth were also monitored during cooling scenarios from 65 to 10°C for 3, 4, 5, 6, and 7 h in vacuum-packed, heat-treated minced turkey. Spores of strain 790-94 were approximately 10-fold more heat resistant at 85°C than those of strain 44071.C05, and strain 790-94 also had a higher temperature growth range in FTG. The higher growth range for a chromosomal enterotoxin-producing CPE+ strain was confirmed using two other strains carrying a chromosomal (NCTC8239) and plasmid (945P) cpe gene. Moreover, strain 790-94 had shorter GOL times at 50°C in turkey and approximately half the generation time compared with strain 44071.C05 at temperatures ≥45°C in both FTG and turkey. Strain 790-94 increased with 0.3, 1.0, 1.7, and 2.0 logs, respectively, during cooling from 65 to 10°C in 4, 5, 6, and 7 h, which was signi cantly higher than for strain 44071.C05. A maximum acceptable cooling time of 5 h between 65 and 10°C is suggested.

Control of Clostridium perfringens spore germination and outgrowth by potassium lactate and sodium diacetate in ham containing reduced sodium chloride

LWT ◽  
2021 ◽  
Vol 137 ◽  
pp. 110395
Author(s):  
Mauricio Redondo-Solano ◽  
Carol Valenzuela-Martinez ◽  
Vijay K. Juneja ◽  
Dennis E. Burson ◽  
Harshavardhan Thippareddi
Keyword(s):  
Sodium Chloride ◽  
Clostridium Perfringens ◽  
Spore Germination ◽  
Potassium Lactate ◽  
Sodium Diacetate

Electrical Properties of Heat-Treated Poly-Lactic Acid

2011 ◽  
Vol 131 (5) ◽  
pp. 395-400 ◽  
Author(s):  
Toru Oi ◽  
Katsuyoshi Shinyama ◽  
Shigetaka Fujita
Keyword(s):  
Lactic Acid ◽  
Electrical Properties ◽  
Poly Lactic Acid ◽  
Heat Treated

scholarly journals The Clostridium perfringens Germinant Receptor Protein GerKC Is Located in the Spore Inner Membrane and Is Crucial for Spore Germination

2013 ◽  
Vol 195 (22) ◽  
pp. 5084-5091 ◽  
Author(s):  
S. Banawas ◽  
D. Paredes-Sabja ◽  
G. Korza ◽  
Y. Li ◽  
B. Hao ◽  
...  
Keyword(s):  
Clostridium Perfringens ◽  
Spore Germination ◽  
Receptor Protein ◽  
Inner Membrane

Susceptibility of Clostridium perfringens to antimicrobials produced by lactic acid bacteria: Reuterin and nisin

Food Control ◽  
2014 ◽  
Vol 44 ◽  
pp. 22-25 ◽  
Author(s):  
Sonia Garde ◽  
Natalia Gómez-Torres ◽  
Marta Hernández ◽  
Marta Ávila
Keyword(s):  
Lactic Acid ◽  
Lactic Acid Bacteria ◽  
Clostridium Perfringens

scholarly journals Germination, Outgrowth, and Vegetative-Growth Kinetics of Dry-Heat-Treated Individual Spores ofBacillusSpecies

2018 ◽  
Vol 84 (7) ◽  
Author(s):  
Lin He ◽  
Zhan Chen ◽  
Shiwei Wang ◽  
Muying Wu ◽  
Peter Setlow ◽  
...  
Keyword(s):  
Heat Treatment ◽  
Spore Germination ◽  
Differential Interference Contrast Microscopy ◽  
Content Type ◽  
Dry Heat ◽  
Heat Treated ◽  
Contrast Microscopy ◽  
Interference Contrast Microscopy ◽  
Dry Heat Treatment ◽  
Kinetics Of

ABSTRACTDNA damage kills dry-heated spores ofBacillus subtilis, but dry-heat-treatment effects on spore germination and outgrowth have not been studied. This is important, since if dry-heat-killed spores germinate and undergo outgrowth, toxic proteins could be synthesized. Here, Raman spectroscopy and differential interference contrast microscopy were used to study germination and outgrowth of individual dry-heat-treatedB. subtilisandBacillus megateriumspores. The major findings in this work were as follows: (i) spores dry-heat-treated at 140°C for 20 min lost nearly all viability but retained their Ca2+-dipicolinic acid (CaDPA) depot; (ii) in most cases, dry-heat treatment increased the average times and variability of all major germination events inB. subtilisspore germination with nutrient germinants or CaDPA, and in one nutrient germination event withB. megateriumspores; (iii)B. subtilisspore germination with dodecylamine, which activates the spore CaDPA release channel, was unaffected by dry-heat treatment; (iv) these results indicate that dry-heat treatment likely damages spore proteins important in nutrient germinant recognition and cortex peptidoglycan hydrolysis, but not CaDPA release itself; and (v) analysis of single spores incubated on nutrient-rich agar showed that while dry-heat-treated spores that are dead can complete germination, they cannot proceed into outgrowth and thus not to vegetative growth. The results of this study provide new information on the effects of dry heat on bacterial spores and indicate that dry-heat sterilization regimens should produce spores that cannot outgrow and thus cannot synthesize potentially dangerous proteins.IMPORTANCEMuch research has shown that high-temperature dry heat is a promising means for the inactivation of spores on medical devices and spacecraft decontamination. Dry heat is known to killBacillus subtilisspores by DNA damage. However, knowledge about the effects of dry-heat treatment on spore germination and outgrowth is limited, especially at the single spore level. In the current work, Raman spectroscopy and differential interference contrast microscopy were used to analyze CaDPA levels in and kinetics of nutrient- and non-nutrient germination of multiple individual dry-heat-treatedB. subtilisandBacillus megateriumspores that were largely dead. The outgrowth and subsequent cell division of these germinated but dead dry-heat-treated spores were also examined. The knowledge obtained in this study will help understand the effects of dry heat on spores both on Earth and in space, and indicates that dry heat can be safely used for sterilization purposes.

scholarly journals Characterization of Clostridium perfringens Spores That Lack SpoVA Proteins and Dipicolinic Acid

2008 ◽  
Vol 190 (13) ◽  
pp. 4648-4659 ◽  
Author(s):  
Daniel Paredes-Sabja ◽  
Barbara Setlow ◽  
Peter Setlow ◽  
Mahfuzur R. Sarker
Keyword(s):  
Water Content ◽  
Uv Radiation ◽  
Clostridium Perfringens ◽  
Spore Germination ◽  
Gastrointestinal Disease ◽  
Dipicolinic Acid ◽  
High Heat ◽  
Wild Type ◽  
Active Growth ◽  
Moist Heat

ABSTRACT Spores of Clostridium perfringens possess high heat resistance, and when these spores germinate and return to active growth, they can cause gastrointestinal disease. Work with Bacillus subtilis has shown that the spore's dipicolinic acid (DPA) level can markedly influence both spore germination and resistance and that the proteins encoded by the spoVA operon are essential for DPA uptake by the developing spore during sporulation. We now find that proteins encoded by the spoVA operon are also essential for the uptake of Ca2+ and DPA into the developing spore during C. perfringens sporulation. Spores of a spoVA mutant had little, if any, Ca2+ and DPA, and their core water content was approximately twofold higher than that of wild-type spores. These DPA-less spores did not germinate spontaneously, as DPA-less B. subtilis spores do. Indeed, wild-type and spoVA C. perfringens spores germinated similarly with a mixture of l-asparagine and KCl (AK), KCl alone, or a 1:1 chelate of Ca2+ and DPA (Ca-DPA). However, the viability of C. perfringens spoVA spores was 20-fold lower than the viability of wild-type spores. Decoated wild-type and spoVA spores exhibited little, if any, germination with AK, KCl, or exogenous Ca-DPA, and their colony-forming efficiency was 103- to 104-fold lower than that of intact spores. However, lysozyme treatment rescued these decoated spores. Although the levels of DNA-protective α/β-type, small, acid-soluble spore proteins in spoVA spores were similar to those in wild-type spores, spoVA spores exhibited markedly lower resistance to moist heat, formaldehyde, HCl, hydrogen peroxide, nitrous acid, and UV radiation than wild-type spores did. In sum, these results suggest the following. (i) SpoVA proteins are essential for Ca-DPA uptake by developing spores during C. perfringens sporulation. (ii) SpoVA proteins and Ca-DPA release are not required for C. perfringens spore germination. (iii) A low spore core water content is essential for full resistance of C. perfringens spores to moist heat, UV radiation, and chemicals.

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