Reproducibility of Salmonella Thermal Resistance Measurements via Multilaboratory Isothermal Inactivation Experiments

2020 ◽  
Vol 83 (4) ◽  
pp. 609-614 ◽  
Author(s):  
IAN M. HILDEBRANDT ◽  
BRADLEY P. MARKS ◽  
NATHAN M. ANDERSON ◽  
ELIZABETH M. GRASSO-KELLEY
Keyword(s):  
Thermal Resistance ◽  
Thermal Inactivation ◽  
Isothermal Treatment ◽  
Inactivation Kinetics ◽  
Sampling Point ◽  
Single Location ◽  
Isothermal Analysis ◽  
D Values ◽  
Log Linear ◽  
Treatment Medium

ABSTRACT Isothermal inactivation experiments often are used to investigate the thermal resistance of pathogens, such as Salmonella, in foods; however, little is known about the reproducibility of such experimental methodologies. The objective of this study was to quantify the reproducibility of Salmonella isothermal resistance results via a six-laboratory comparison. Inoculation was performed at a single location and then distributed to each laboratory for isothermal analysis. Salmonella Agona 447967 was inoculated into oat flour, re-equilibrated to a water activity (aw) of 0.45, and then packaged and distributed to each laboratory. Before conducting the inactivation trials, each laboratory was required to verify the inoculated product's aw, enumerate Salmonella population levels, and verify that the isothermal treatment medium was at the target temperature (80°C). All laboratories were required to process at least three replications, collect at least six sample time points with three subsamples at each sampling point, enumerate survivors using an identical plating methodology and media, and verify that the temperature did not substantially change during isothermal treatment. The log-linear model was fit to the Salmonella survivor data, and the resultant D-values were statistically compared via Welch's t test (α = 0.05). Two significant differences in thermal inactivation kinetics were identified as potentially resulting from suspected methodology deviations. Two of the inoculated batches distributed for analysis yielded significantly lower D-values, which likely resulted from a deviation in the inoculation procedures. One laboratory yielded significantly lower D-values, which was likely the result of temperature deviations. Overall, excluding the D-values resulting from deviations, the inactivation results were reproducible, yielding D-values of 30.2 ± 3 min. These results indicate that isothermal inactivation results can be reproducible but that even minor methodology deviations can substantially affect measured Salmonella thermal resistance. HIGHLIGHTS

Effects of Inoculation Procedures on Variability and Repeatability of Salmonella Thermal Resistance in Wheat Flour

2016 ◽  
Vol 79 (11) ◽  
pp. 1833-1839 ◽  
Author(s):  
IAN M. HILDEBRANDT ◽  
BRADLEY P. MARKS ◽  
ELLIOT T. RYSER ◽  
ROSSANA VILLA-ROJAS ◽  
JUMING TANG ◽  
...  
Keyword(s):  
Thermal Resistance ◽  
Water Activity ◽  
Wheat Flour ◽  
Thermal Inactivation ◽  
Culture Method ◽  
Isothermal Treatment ◽  
Inactivation Kinetics ◽  
Inoculation Methods ◽  
Low Moisture Foods ◽  
D Values

ABSTRACT Limited prior research has shown that inoculation methods affect thermal resistance of Salmonella in low-moisture foods; however, these effects and their repeatability have not been systematically quantified. Consequently, method variability across studies limits utility of individual data sets and cross-study comparisons. Therefore, the objective was to evaluate the effects of inoculation methodologies on stability and thermal resistance of Salmonella in a low-moisture food (wheat flour), and the repeatability of those results, based on data generated by two independent laboratories. The experimental design consisted of a cross-laboratory comparison, both conducting isothermal Salmonella inactivation studies in wheat flour (~0.45 water activity, 80°C), utilizing five different inoculation methods: (i) broth-based liquid inoculum, (ii) lawn-based liquid inoculum, (iii) lawn-based pelletized inoculum, (iv) direct harvest of lawn culture with wheat flour, and (v) fomite transfer of a lawn culture. Inoculated wheat flour was equilibrated ~5 days to ~0.45 water activity and then was subjected to isothermal treatment (80°C) in aluminum test cells. Results indicated that inoculation method impacted repeatability, population stability, and inactivation kinetics (α = 0.05), regardless of laboratory. Salmonella inoculated with the broth-based liquid inoculum method and the fomite transfer of a lawn culture method exhibited instability during equilibration. Lawn-based cultures resulted in stable populations prior to thermal treatment; however, the method using direct harvest of lawn culture with wheat flour yielded different D-values across the laboratories (α = 0.05), which was attributed to larger potential impact of operator variability. The lawn-based liquid inoculum and the lawn-based pelletized inoculum methods yielded stable inoculation levels and repeatable D-values (~250 and ~285 s, respectively). Also, inoculation level (3 to 8 log CFU/g) did not affect D-values (using the lawn-based liquid inoculum method). Overall, the results demonstrate that inoculation methods significantly affect Salmonella population kinetics and subsequent interpretation of thermal inactivation data for low-moisture foods.

Enhanced Thermal Resistance of Salmonella in Marinated Whole Muscle Compared with Ground Pork

2010 ◽  
Vol 73 (2) ◽  
pp. 372-375 ◽  
Author(s):  
ADRIANA VELASQUEZ ◽  
TASHA J. BRESLIN ◽  
BRADLEY P. MARKS ◽  
ALICIA ORTA-RAMIREZ ◽  
NICOLE O. HALL ◽  
...  
Keyword(s):  
Thermal Resistance ◽  
Physical State ◽  
Thermal Inactivation ◽  
Heating Temperature ◽  
Inactivation Kinetics ◽  
Pork Products ◽  
Ground Pork ◽  
Lag Times ◽  
Log Linear ◽  
Whole Muscle

The internal muscle environment may enhance thermal resistance of bacterial pathogens. Based on the migration of pathogens into whole muscle products during marination, the validity of current thermal inactivation models for whole muscle versus ground products has been questioned. Consequently, the objective of this work was to compare thermal resistance of Salmonella in whole muscle versus ground pork. Irradiated samples of whole and ground pork loin (5.5 to 7.5 g) were exposed to a Salmonella-inoculated (108 CFU/ml) marinade (eight serovar cocktail) for 20 min, placed in sterile brass tubes (12.7 mm diameter), sealed, and heated isothermally at 55, 58, 60, 62, or 63°C, and surviving salmonellae were enumerated on Petrifilm aerobic count plates. The thermal lag times and initial bacterial counts were similar for both whole muscle and ground samples (P > 0.05), with all samples having equivalent compositions, inocula, and thermal histories. Heating temperature and physical state of the meat (whole versus ground muscle) affected Salmonella inactivation, with greater thermal resistance observed in whole than in ground muscle (P < 0.05). Assuming log-linear inactivation kinetics, Salmonella was 0.64 to 2.96 times more heat resistant in whole muscle than in ground pork. Therefore, thermal process validations for pork products should also account for the physical state of the product to ensure microbial safety.

Desiccation and Thermal Resistance of Escherichia coli O121 in Wheat Flour

2019 ◽  
Vol 82 (8) ◽  
pp. 1308-1313 ◽  
Author(s):  
QUINCY J. SUEHR ◽  
NATHAN M. ANDERSON ◽  
SUSANNE E. KELLER
Keyword(s):  
Escherichia Coli ◽  
Thermal Resistance ◽  
Wheat Flour ◽  
Shiga Toxin ◽  
Salmonella Enteritidis ◽  
Thermal Inactivation ◽  
The United States ◽  
Inactivation Kinetics ◽  
E Coli ◽  
D Values

ABSTRACT Non-O157 Shiga toxin–producing Escherichia coli infections have recently been associated with wheat flour on two separate accounts in the United States and Canada. However, there is little information regarding the thermal resistance and longevity of non-O157 Shiga toxin–producing Escherichia coli during storage in low-moisture environments. The objectives of this study were to determine the thermal inactivation kinetics of E. coli O121 in wheat flour and to compare the thermal inactivation rates with those of other pathogens. Wheat flour, inoculated with E. coli O121, was equilibrated at 25°C to a water activity of 0.45 in a humidity-controlled conditioning chamber. Inoculated samples were treated isothermally at 70, 75, and 80°C, and posttreatment population survivor ratios were determined by plate counting. D- and z-values calculated with a log-linear model, were compared with those obtained in other studies. At 70, 75, and 80°C, the D-values for E. coli O121 were 18.16 ± 0.96, 6.47 ± 0.50, and 4.58 ± 0.40 min, respectively, and the z-value was 14.57 ± 2.21°C. Overall, E. coli O121 was observed to be slightly less thermally resistant than what has been previously reported for Salmonella Enteritidis PT30 in wheat flour as measured under the same conditions with the same methods.

Inactivation Kinetics of Pathogens during Thermal Processing in Acidified Broth and Tomato Purée (pH 4.5)

2017 ◽  
Vol 80 (12) ◽  
pp. 2014-2021 ◽  
Author(s):  
Evann L. Dufort ◽  
Jonathan Sogin ◽  
Mark R. Etzel ◽  
Barbara H. Ingham
Keyword(s):  
Thermal Inactivation ◽  
Weibull Model ◽  
Inactivation Kinetics ◽  
E Coli ◽  
Log Reduction ◽  
Tomato Puree ◽  
Weibull Models ◽  
Canned Foods ◽  
D Values ◽  
Log Linear

ABSTRACT Thermal inactivation kinetics for single strains of Shiga toxin–producing Escherichia coli (STEC), Listeria monocytogenes, and Salmonella enterica were measured in acidified tryptic soy broth (TSB; pH 4.5) heated at 54°C. Inactivation curves also were measured for single-pathogen five-strain cocktails of E. coli O157:H7, L. monocytogenes, and S. enterica heated in tomato purée (pH 4.5) at 52, 54, 56, and 58°C. Inactivation curves were fit using log-linear and nonlinear (Weibull) models. The Weibull model yields the time for a 5-log reduction (t*) and a curve shape parameter (β). Decimal reduction times (D-values) and thermal resistance constants (z-values) from the two models were compared by defining t* = 5D* for the Weibull model. When the log-linear and Weibull models match at the 5-log reduction time, then t* = 5D* = 5D and D = D*. In 18 of 20 strains heated in acidified TSB, D and D* for the two models were not significantly different, although nonlinearity was observed in 35 of 60 trials. Similarly, in 51 of 52 trials for pathogen cocktails heated in tomato purée, D and D* were not significantly different, although nonlinearity was observed in 31% of trials. At a given temperature, D-values for S. enterica << L. monocytogenes < E. coli O157:H7 in tomato purée (pH 4.5). When using the two models, z-values calculated from the D-values were not significantly different for a given pathogen. Across all pathogens, z-values for E. coli O157:H7 and S. enterica were not different but were significantly lower than the z-values for L. monocytogenes. These results are useful for supporting process filings for tomato-based acidified food products with pH 4.5 and below and are relevant to small processors of tomato-based acidified canned foods who do not have the resources to conduct research on and validate pathogen lethality.

Relationships of Water Activity and Moisture Content to the Thermal Inactivation Kinetics of Salmonella in Low-Moisture Foods

2019 ◽  
Vol 82 (6) ◽  
pp. 963-970 ◽  
Author(s):  
FRANCISCO J. GARCES-VEGA ◽  
ELLIOT T. RYSER ◽  
BRADLEY P. MARKS
Keyword(s):  
Moisture Content ◽  
Water Activity ◽  
Thermal Inactivation ◽  
Inactivation Kinetics ◽  
Temperature Dependent ◽  
Pathogen Control ◽  
Low Moisture Foods ◽  
D Values ◽  
Log Linear ◽  
Kinetics Of

ABSTRACT The enhanced thermal resistance of Salmonella in low-moisture foods (LMFs) presents a challenge when validating pathogen control processes. Product water is recognized as a controlling factor in thermal inactivation of Salmonella in or on LMFs, such as almonds. Water activity (aw) describes the state of water in a product; however, aw is temperature dependent and characterized by hysteresis between sorption states. Moisture content (%MC) describes the amount of water in a product; it is not temperature dependent and might be a more convenient metric than aw to account for water in thermal inactivation processes. To test these two metrics independently, Salmonella-inoculated almonds were equilibrated to two %MC levels but the same aw and to two aw levels but the same %MC. Equilibrated products were vacuum packaged and thermally treated in a water bath at 80°C. Survivors were recovered and enumerated. The resulting inactivation curves were used to fit the log-linear inactivation model, and the inactivation kinetics were compared. D-values ranged from 15.7 to 18.0 min, and the root mean square error was 0.25 to 0.69 log CFU/g. No differentiated (P > 0.05) effect attributable preferentially to aw or %MC was seen in the inactivation kinetics. The separate effects of aw and %MC on the inactivation kinetics of Salmonella in LMFs remain inconclusive, but analyses of data from prior studies strongly suggested an effect of sorption state. Further analysis is needed to identify which metric is best for modeling and validating thermal inactivation processes. HIGHLIGHTS

Thermal Inactivation Kinetics of Three Heat-Resistant Salmonella Strains in Whole Liquid Egg

2019 ◽  
Vol 82 (9) ◽  
pp. 1465-1471 ◽  
Author(s):  
JOSHUA B. GURTLER ◽  
VIJAY K. JUNEJA ◽  
DEANA R. JONES ◽  
ANUJ PUROHIT
Keyword(s):  
Survival Data ◽  
Thermal Inactivation ◽  
Inactivation Kinetics ◽  
Glass Capillary ◽  
Circulating Water ◽  
Log Reduction ◽  
Heat Resistant ◽  
Weibull Models ◽  
D Values ◽  
Log Linear

ABSTRACT The heat resistance of three heat-resistant strains of Salmonella was determined in whole liquid egg (WLE). Inoculated samples in glass capillary tubes were completely immersed in a circulating water bath and held at 56, 58, 60, 62, and 64°C for predetermined lengths of time. The recovery medium was tryptic soy agar with 0.1% sodium pyruvate and 50 ppm of nalidixic acid. Survival data were fitted using log-linear, log-linear with shoulder, and Weibull models using GInaFiT version 1.7. Based on the R2 and mean square error, the log-linear with shoulder and Weibull models consistently produced a better fit to Salmonella survival curves obtained at these temperatures. Contaminated WLE must be heated at 56, 60, and 64°C for at least 33.2, 2.7, and 0.31 min, respectively, to achieve a 4-log reduction of Salmonella; 39.0, 3.1, and 0.34 min, respectively, for a 5-log reduction; and 45.0, 3.5, and 0.39 min, respectively, for a 6-log reduction. The z-values calculated from the D-values were 3.67 and 4.18°C for the log-linear with shoulder and Weibull models, respectively. Thermal death times presented in this study will be beneficial for WLE distributors and regulatory agencies when designing pasteurization processes to effectively eliminate Salmonella in WLE, thereby ensuring the microbiological safety of the product.

A Comparison of Three Methods for Determining Thermal Inactivation Kinetics: A Case Study on Salmonellaenterica in Whole Milk Powder

2020 ◽  
Author(s):  
Soon Kiat Lau ◽  
Xinyao Wei ◽  
Nina Kirezi ◽  
Rajendra Panth ◽  
Arena See ◽  
...  
Keyword(s):  
Thermal Resistance ◽  
Thermal Inactivation ◽  
Milk Powder ◽  
Water Bath ◽  
Isothermal Treatment ◽  
Inactivation Kinetics ◽  
Model Parameters ◽  
Treatment Methods ◽  
Whole Milk ◽  
Whole Milk Powder

Different methods for determining the thermal inactivation kinetics of microorganisms could result in discrepancies of the resulting thermal resistance values. This study determined the thermal resistance of Salmonella in whole milk powder using three methods (Thermal death time (TDT) disk in water bath, pouches in water bath, and TDT Sandwich). Samples from three separate production lots of whole milk powder were inoculated with a five strain Salmonella cocktail and equilibrated to 0.20 a w . The samples were then subjected to three isothermal treatment methods at 75, 80, or 85 °C. Samples were taken out at six timepoints and enumerated for survivors. The inactivation data were fitted to two consolidated models consisting of two primary models (log-linear and Weibull) and one secondary model (Bigelow). Normality testing indicated that all the model parameters were normally distributed. None of the model parameters for both consolidated models were significantly different (α=0.05). The amount of inactivation during the come-up time phase was also not significantly different among the methods (α=0.05). In terms of magnitude, however, the TDT Sandwich showed less inactivation during the come-up time phase and overall less variation in model parameters. The survivor data from all three methods were combined and fitted to both consolidated models, with the Weibull having lower root mean square error and a better fit according to corrected Akaike’s Information Criterion. These results suggest that the three thermal treatment methods are not significantly different from each other and are interchangeable, at least in the case of Salmonella in whole milk powder. Comparisons with more methods, other microorganisms, and larger varieties of food products using the same framework presented in this study could provide guidance for standardizing thermal inactivation kinetics studies for microorganisms in foods.

scholarly journals Modelling the thermal inactivation of viruses from the Coronaviridae family in suspensions or on surfaces with various relative humidities

2020 ◽  
Author(s):  
Laurent Guillier ◽  
Sandra Martin-Latil ◽  
Estelle Chaix ◽  
Anne Thébault ◽  
Nicole Pavio ◽  
...  
Keyword(s):  
Relative Humidity ◽  
Thermal Inactivation ◽  
Inactivation Kinetics ◽  
Virus Infectivity ◽  
Virus Inactivation ◽  
Public Authorities ◽  
Contact Transmission ◽  
D Values ◽  
Major Factors ◽  
The Impact

AbstractTemperature and relative humidity are major factors determining virus inactivation in the environment. This article reviews inactivation data of coronaviruses on surfaces and in liquids from published studies and develops secondary models to predict coronaviruses inactivation as a function of temperature and relative humidity. A total of 102 D-values (time to obtain a log10 reduction of virus infectivity), including values for SARS-CoV-2, were collected from 26 published studies. The values obtained from the different coronaviruses and studies were found to be generally consistent. Five different models were fitted to the global dataset of D-values. The most appropriate model considered temperature and relative humidity. A spreadsheet predicting the inactivation of coronaviruses and the associated uncertainty is presented and can be used to predict virus inactivation for untested temperatures, time points or new coronavirus strains.ImportanceThe prediction of the persistence of SARS-CoV-2 on fomites is essential to investigate the importance of contact transmission. This study collects available information on inactivation kinetics of coronaviruses in both solid and liquid fomites and creates a mathematical model for the impact of temperature and relative humidity on virus persistence. The predictions of the model can support more robust decision-making and could be useful in various public health contexts. Having a calculator for the natural clearance of SARS-CoV-2 depending on temperature and relative humidity could be a valuable operational tool for public authorities.

Thermal Resistance of Aeromonas hydrophila in Liquid Whole Egg†

1997 ◽  
Vol 60 (3) ◽  
pp. 231-236 ◽  
Author(s):  
JAMES D. SCHUMAN ◽  
BRIAN W. SHELDON ◽  
PEGGY M. FOEGEDING
Keyword(s):  
Thermal Resistance ◽  
Aeromonas Hydrophila ◽  
Animal Origin ◽  
Inactivation Kinetics ◽  
Processing Plant ◽  
Decimal Reduction Time ◽  
Heat Resistant ◽  
Liquid Whole Egg ◽  
D Values ◽  
Whole Egg

Aeromonas hydrophila (AH) is a psychrotrophic spoilage bacterium and potential pathogen which has been isolated from a variety of refrigerated foods of animal origin, including raw milk, red meat, poultry, and commercially broken raw liquid whole egg (LWE). Decimal reduction times (D values) of 4 strains of AH (1 egg isolate, 2 egg processing plant isolates, 1 ATCC type strain) were determined in LWE using an immersed sealed capillary tube (ISCT) procedure. Initial populations (7.0 to 8.3 log CFU/tube in 0.05 ml LWE) were heated at 48, 51, 54, 57, and 60°C, and survivors were plated onto starch ampicillin agar (48 h at 28°C). D values ranged from 3.62 to 9.43 min (at 48°C) to 0.026 to 0.040 min (at 60°C). Both processing plant isolates were more heat resistant than the ATCC strain. Decimal reduction time curves (r2 ≤ 0.98) yielded ZD values of 5.02 to 5.59°C, similar to those for other non-spore-forming bacteria. D values of the most heat resistant AH strain were also determined in LWE at 48, 51, and 54°C using a conventional capped test tube procedure (10 ml/tube). Cells heated in test tubes yielded nonlinear (tailing) survivor curves and larger (P ≤ 0.05) apparent D values at each temperature than those obtained using the ISCT method. This study provides the first thermal resistance data for AH in LWE and the first evidence that straight-line semilogarithmic thermal inactivation kinetics may be demonstrated for Aeromonas using the ISCT procedure.

Determination of the Thermal Inactivation Kinetics of Listeria monocytogenes, Salmonella enterica, and Escherichia coli O157:H7 and non-O157 in Buffer and a Spinach Homogenate

2015 ◽  
Vol 78 (8) ◽  
pp. 1467-1471 ◽  
Author(s):  
EMEFA ANGELICA MONU ◽  
MALCOND VALLADARES ◽  
DORIS H. D'SOUZA ◽  
P. MICHAEL DAVIDSON
Keyword(s):  
Escherichia Coli ◽  
Listeria Monocytogenes ◽  
Salmonella Enterica ◽  
Thermal Inactivation ◽  
Inactivation Kinetics ◽  
Mild Heat ◽  
Log Reduction ◽  
D Values ◽  
Heat Processes ◽  
Kinetics Of

Produce has been associated with a rising number of foodborne illness outbreaks. While much produce is consumed raw, some is treated with mild heat, such as blanching or cooking. The objectives of this research were to compare the thermal inactivation kinetics of Listeria monocytogenes, Salmonella enterica, Shiga toxin–producing Escherichia coli (STEC) O157:H7, and non-O157 STEC in phosphate-buffered saline (PBS; pH 7.2) and a spinach homogenate and to provide an estimate of the safety of mild heat processes for spinach. Five individual strains of S. enterica, L. monocytogenes, STEC O157:H7, and non-O157 STEC were tested in PBS in 2-ml glass vials, and cocktails of the organisms were tested in blended spinach in vacuum-sealed bags. For Listeria and Salmonella at 56 to 60°C, D-values in PBS ranged from 4.42 ± 0.94 to 0.35 ± 0.03 min and 2.11 ± 0.14 to 0.16 ± 0.03 min, respectively. D-values at 54 to 58°C were 5.18 ± 0.21 to 0.53 ± 0.04 min for STEC O157:H7 and 5.01 ± 0.60 to 0.60 ± 0.13 min for non-O157 STEC. In spinach at 56 to 60°C, Listeria D-values were 11.77 ± 2.18 to 1.22 ± 0.12 min and Salmonella D-values were 3.51 ± 0.06 to 0.47 ± 0.06 min. D-values for STEC O157:H7 and non-O157 STEC were 7.21 ± 0.17 to 1.07 ± 0.11 min and 5.57 ± 0.38 to 0.99 ± 0.07 min, respectively, at 56 to 60°C. In spinach, z-values were 4.07 ± 0.16, 4.59 ± 0.26, 4.80 ± 0.92, and 5.22 ± 0.20°C for Listeria, Salmonella, STEC O157:H7, and non-O157 STEC, respectively. Results indicated that a mild thermal treatment of blended spinach at 70°C for less than 1 min would result in a 6-log reduction of all pathogens tested. These findings may assist the food industry in the design of suitable mild thermal processes to ensure food safety.

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