Inactivation of Desiccation-Resistant Salmonella on Apple Slices following Treatment with ε-Polylysine, Sodium Bisulfate or Peracetic Acid and Subsequent Dehydration.

Salmonella is capable of surviving dehydration within various foods, such as dried fruit. Dried fruit, including apple slices, have been the subject of product recalls due to contamination with Salmonella. A study was conducted to determine the fate of Salmonella on apple slices, following immersion in three antimicrobial solutions (viz., ε-polylysine [epsilon-polylysine or EP], sodium bisulfate [SBS] or peracetic acid [PAA]), and subsequent hot air dehydration. Gala apples were aseptically cored and sliced into 0.4 cm thick rings, bisected, and inoculated with a five-strain composite of desiccation-resistant Salmonella, to a population of 8.28 log CFU/slice. Slices were then immersed for 2 min in various concentrations of antimicrobial solutions, including EP (0.005, 0.02, 0.05 and 0.1%), SBS (0.05, 0.1, 0.2 and 0.3%), PAA (18 or 42 ppm), or varying concentrations of PAA + EP, and then dehydrated at 60°C for 5 h. Salmonella populations in positive control samples (inoculated apple slices washed in sterile water) declined by 2.64 log after drying. In the present study, inactivation of Salmonella, following EP and SBS treatments, increased with increasing concentrations, with maximum reductions of 3.87 and 6.20 log (with 0.1 and 0.3% of the two compounds, respectively). Based on preliminary studies, EP concentrations greater than 0.1% did not result in lower populations of Salmonella. Pretreatment washes with either 18 or 42 ppm of PAA inactivated Salmonella populations by 4.62 and 5.63 log, respectively, following desiccation. Combining PAA with up to 0.1% EP induced no greater population reductions of Salmonella than washing with PAA alone. The addition of EP to PAA solutions appeared to destabilize PAA concentrations, reducing its biocidal efficacy. These results may provide antimicrobial pre-drying treatment alternatives to promote the reduction of Salmonella during commercial or consumer hot air drying of apple slices.

Practice and Progress: Updates on Outbreaks, Advances in Research, and Processing Technologies for Low-moisture Food Safety.

Large, renowned outbreaks associated with low-moisture foods (LMFs) bring to light some of the potential, inherent risks that accompany foods with long shelf lives if pathogen contamination occurs. Subsequently, in 2013, Beuchat et al. (2013) noted the increased concern regarding these foods, specifically noting examples of persistence and resistance of pathogens in low-water activity foods (LWAFs), prevalence of pathogens in LWAF processing environments, and sources of and preventive measures for contamination of LWAFs. For the last decade, the body of knowledge related to LMF safety has exponentially expanded. This growing field and interest in LMF safety have led researchers to delve into survival and persistence studies, revealing that some foodborne pathogens can survive in LWAFs for months to years. Research has also uncovered many complications of working with foodborne pathogens in desiccated states, such as inoculation methods and molecular mechanisms that can impact pathogen survival and persistence. Moreover, outbreaks, recalls, and developments in LMF safety research have created a cascading feedback loop of pushing the field forward, which has also led to increased attention on how industry can improve LMF safety and raise safety standards. Scientists across academia, government agencies, and industry have partnered to develop and evaluate innovate thermal and nonthermal technologies to use on LMFs, which are described in the presented review. The objective of this review was to describe aspects of the extensive progress made by researchers and industry members in LMF safety, including lessons-learned about outbreaks and recalls, expansion of knowledge base about pathogens that contaminate LMFs, and mitigation strategies currently employed or in development to reduce food safety risks associated with LMFs.

Interlaboratory Evaluation of Enterococcus faecium NRRL B-2354 as a Salmonella Surrogate for Validating Thermal Treatment of Multiple Low-Moisture Foods.

ABSTRACT: This multi-institutional study assessed the efficacy of Enterococcus faecium NRRL B-2354 as a nonpathogenic Salmonella surrogate for thermal processing of nonfat dry milk powder, peanut butter, almond meal, wheat flour, ground black pepper, and date paste. Each product was analyzed by two laboratories (five independent laboratories total), with the lead laboratory inoculating (E. faecium or a five-strain Salmonella enterica serovar cocktail of Agona, Reading, Tennessee, Mbandaka, and Montevideo) and equilibrating the product to the target water activity before shipping. Both laboratories subjected samples to three isothermal treatments (between 65 and 100°C). A log-linear and Bigelow model was fit to survivor data via one-step regression. On the basis of D80°C values estimated from the combined model, E. faecium was more thermally resistant (P < 0.05) than Salmonella in nonfat dry milk powder (DEf-80°C, 100.2 ± 5.8 min; DSal-80°C, 28.9 ± 1.0 min), peanut butter (DEf-80°C, 133.5 ± 3.1 min; DSal-80°C, 57.6 ± 1.5 min), almond meal (DEf-80°C, 34.2 ± 0.4 min; DSal-80°C, 26.1 ± 0.2 min), ground black pepper (DEf-80°C, 3.2 ± 0.8 min; DSal-80°C, 1.5 ± 0.1 min), and date paste (DEf-80°C, 1.5 ± 0.0 min; DSal-80°C, 0.5 ± 0.0 min). Although the combined laboratory D80°C for E. faecium was lower (P < 0.05) than for Salmonella in wheat flour (DEf-80°C, 9.4 ± 0.1 min; DSal-80°C, 10.1 ± 0.2 min), the difference was ∼7%. The zT values for Salmonella in all products and for E. faecium in milk powder, almond meal, and date paste were not different (P > 0.05) between laboratories. Therefore, this study demonstrated the impact of standardized methodologies on repeatability of microbial inactivation results. Overall, E. faecium NRRL B-2354 was more thermally resistant than Salmonella, which provides support for utilizing E. faecium as a surrogate for validating thermal processing of multiple low-moisture products. However, product composition should always be considered before making that decision.

Predictive Microbial Modeling of Enterococcus faecium NRRL B-2354 Inactivation during Baking of a Multicomponent Low-Moisture Food.

ABSTRACT: The use of baking ovens as a microbial kill step should be validated based on results of thermal inactivation models. Although traditional isothermal models may not be appropriate for these dynamic processes, they are being used by the food industry. Previous research indicates that the impact of additional process conditions, such as humidity, should be considered when validating thermal processes for the control of microbial hazards in low-moisture foods. In this study, the predictive performance of traditional and modified thermal inactivation kinetic models accounting for process humidity were assessed for predicting inactivation of Enterococcus faecium NRRL B-2354 in a multi-ingredient composite food during baking. Ingredients (milk powder, protein powder, peanut butter, and whole wheat flour) were individually inoculated to achieve ∼6 log CFU/g, equilibrated to a water activity of 0.25, and then mixed to form a cookie dough. An isothermal inactivation study was conducted for the dough to obtain traditional D- and z-values (n = 63). In a separate experiment, cookies were baked under four dynamic heating conditions: 135°C, high humidity; 135°C, low humidity; 150°C, high humidity; and 150°C, low humidity. Process humidity measurements; time-temperature profiles for the product core, surface, and bulk air; and microbial survivor ratios were collected for the four conditions at six residence times (n = 144). The traditional isothermal model had a high root mean square error (RMSE) of 856.51 log CFU/g, significantly overpredicting bacterial inactivation during the process. The modified model accounting for the dynamic time-temperature profile and process humidity data was a better predictor with an RMSE of 0.55 log CFU/g. These results indicate the importance of accounting for additional process parameters in baking inactivation models and that model performance can be improved by utilizing model parameters obtained directly from industrial-scale experimental data.

Evaluation of Hot-Air Drying To Inactivate Salmonella and Enterococcus faecium on Apple Pieces.

ABSTRACT: Hot-air drying processes are used to provide specific quality attributes to products, such as dehydrated apple pieces. To comply with the U.S. Food and Drug Administration Food Safety Modernization Act, there is a need to understand microbial lethality during these processes. The objective of this study was to determine the level of inactivation provided by hot-air drying on a Salmonella cocktail inoculated onto apple cubes and to evaluate the performance of Enterococcus faecium as a surrogate. A cocktail of Salmonella serovars (Agona, Tennessee, Montevideo, Mbandaka, and Reading) and E. faecium were individually inoculated onto cored, peeled Gala apple cubes at 9.2 ± 0.3 and 8.8 ± 0.1 log CFU per sample, respectively. Apple cubes were dried at 104 or 135°C in ∼1.5-kg batches using a hot-air dryer with a vertically directed heat source and without mixing. Three subsamples, consisting of four inoculated cubes, were enumerated at each time point (n ≥ 5) from multiple product bed depths. Water activity decreased throughout the duration of the study, with samples drying faster at 135 than 104°C. Samples at the bottom bed depth, closer to the heat source, dried faster than those at the higher bed depth, regardless of temperature. Significant microbial inactivation was not seen immediately. It took >10 min at the bottom bed depth or >40 min of drying at the top bed depth, regardless of temperature (P < 0.05). By the end of drying, average Salmonella inactivation of greater than 5 log CFU per sample was achieved. At temperature conditions evaluated, E. faecium inactivation was slower than Salmonella, indicating that it would likely serve as a good surrogate for in-plant validation studies. Case hardening did not inhibit microbial inactivation in the conditions tested. Hot-air drying under the conditions evaluated may provide a preventive control in the production of dehydrated products, such as apples.

Reproducibility of Salmonella Thermal Resistance Measurements via Multilaboratory Isothermal Inactivation Experiments.

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.

Thermal Inactivation of Salmonella Agona in Low-Water Activity Foods: Predictive Models for the Combined Effect of Temperature, Water Activity, and Food Component.

Salmonella can survive in low-moisture, high-protein, and high-fat foods for several years. Despite nationwide outbreaks and recalls due to the presence of Salmonella in low-moisture foods, information on thermal inactivation of Salmonella in these products is limited. This project evaluated the impact of water activity (aw), temperature, and food composition on thermal inactivation of Salmonella enterica serovar Agona in defined high-protein and high-fat model food matrices. Each matrix was inoculated with Salmonella Agona and adjusted to obtain a target aw, ranging from 0.50 to 0.98. Samples were packed into aluminum test cells and heated (52 to 90°C) under isothermal conditions. Survival of Salmonella Agona was detected on tryptic soy agar with 0.6% yeast extract. Complex influences by food composition, aw, and temperature resulted in significantly different ( P < 0.05) thermal resistance of Salmonella for the conditions tested. It was estimated that the same point temperatures at which the D-values of the two matrices at each aw (0.63, 0.73, 0.81, and 0.90) were identical were 79.48, 71.28, 69.62, and 38.42°C, respectively. Above these temperatures, the D-values in high-protein matrices were larger than the D-values in high-fat matrices at each aw. Below these temperatures, the inverse relationship was observed. A correlation between temperature and aw existed on the basis of the level of fat or protein in the food, showing that these compositional factors must be accounted for when predicating thermal inactivation of Salmonella in foods.

Fate of Salmonella throughout Production and Refrigerated Storage of Tahini.

Tahini, a low-moisture food that is made from sesame seeds, has been implicated in outbreaks of salmonellosis. In this study, the fate of Salmonella was determined through an entire process for the manufacture of tahini, including a 24-h seed soaking period before roasting, subsequent grinding, and storage at refrigeration temperature. Salmonella populations increased by more than 3 log CFU/g during a 24-h soaking period, reaching more than 7 log CFU/g. Survival of Salmonella during roasting at three temperatures, 95, 110, and 130°C, was assessed using seeds on which Salmonella was grown. Salmonella survival was impacted both by temperature and the water activity (aw) at the beginning of the roasting period. When roasted at 130°C with a high initial aw (≥0.90) and starting Salmonella populations of ∼8.5 log CFU/g, populations quickly decreased below detection limits within the first 10 min. However, when the seeds were reduced to an aw of 0.45 before roasting at the same temperature, 3.5 log CFU/g remained on the seeds after 60 min. In subsequent storage studies, seeds were roasted at 130°C for 15 min before processing into tahini. For the storage studies, tahini was inoculated using two methods. The first method used seeds on which Salmonella was first grown before roasting. In the second method, Salmonella was inoculated into the tahini after manufacture. All tahini was stored for 119 days at 4°C. No change in Salmonella populations was recorded for tahini throughout the entire 119 days regardless of the inoculation method used. These combined results indicate the critical importance of aw during a roasting step during tahini manufacture. Salmonella that survive roasting will likely remain viable throughout the normal shelf life of tahini.

Dry Transfer Inoculation of Low-Moisture Spices Containing Antimicrobial Compounds.

Inoculation of a food product for use in subsequent validation studies typically makes use of a high concentration cocktail of microorganisms suspended in aqueous media. However, this inoculation method may prove difficult particularly when the food product is a low-moisture food containing antimicrobial compounds, such as some dried spices. In this study, a dry transfer method for inoculation of clove powder, oregano leaves, ginger powder, and ground black pepper with a five-serovar cocktail of Salmonella was developed and compared with a traditional aqueous inoculation procedure. Spices were inoculated at three levels, 10, 8, and 6 log CFU/g, by using both an aqueous suspension of Salmonella and a dry transfer of Salmonella from previously inoculated silica beads. At the highest inoculation level, the dry transfer method resulted in a significantly higher microbial load (P < 0.05) for ground cloves and oregano, but not for ginger and ground black pepper. At the intermediate inoculation level, differences were apparent only for ginger and black pepper. Inoculation levels of 6 log CFU/g resulted in recoveries below detection limits for both methods of inoculation. Additional examination on the survival of Salmonella on silica beads after inoculation and in clove powder after dry transfer from silica beads showed linear rates of decline, with a rate of -0.011 log CFU/g/day for beads and -0.015 log CFU/g/day for clove powder. The results suggest that dry transfer of Salmonella via inoculated silica beads is a viable alternative when traditional aqueous inoculation is not feasible.

Salmonella Inactivation During Extrusion of an Oat Flour Model Food.

Little research exists on Salmonella inactivation during extrusion processing, yet many outbreaks associated with low water activity foods since 2006 were linked to extruded foods. The aim of this research was to study Salmonella inactivation during extrusion of a model cereal product. Oat flour was inoculated with Salmonella enterica serovar Agona, an outbreak strain isolated from puffed cereals, and processed using a single-screw extruder at a feed rate of 75 kg/h and a screw speed of 500 rpm. Extrudate samples were collected from the barrel outlet in sterile bags and immediately cooled in an ice-water bath. Populations were determined using standard plate count methods or a modified most probable number when populations were low. Reductions in population were determined and analyzed using a general linear model. The regression model obtained for the response surface tested was Log (NR /NO ) = 20.50 + 0.82T - 141.16aw - 0.0039T2 + 87.91aw2 (R2 = 0.69). The model showed significant (p < 0.05) linear and quadratic effects of aw and temperature and enabled an assessment of critical control parameters. Reductions of 0.67 ± 0.14 to 7.34 ± 0.02 log CFU/g were observed over ranges of aw (0.72 to 0.96) and temperature (65 to 100 °C) tested. Processing conditions above 82 °C and 0.89 aw achieved on average greater than a 5-log reduction of Salmonella. Results indicate that extrusion is an effective means for reducing Salmonella as most processes commonly employed to produce cereals and other low water activity foods exceed these parameters. Thus, contamination of an extruded food product would most likely occur postprocessing as a result of environmental contamination or through the addition of coatings and flavorings.