Inactivation of Alicyclobacillus contaminans spores by dielectric barrier discharge plasma and its biological mechanism

Over the years, as an emerging technology, cold plasma (CP) has been widely used in the preservation of agricultural products. Alicyclobacillus spp. are spore-forming bacteria and difficult to inactivate. They adversely affect the economic value of agricultural products. Nevertheless, there are still few studies related to the inactivation of spores using CP. Herein, the inactivation effect and mechanism of dielectric barrier discharge plasma (DBDP) on Alicyclobacillus contaminans (A. contaminans) spores in phosphate-buffered saline and clarified apple juice were investigated. Plasma treatment at 75 V for 9 min achieved 99% inactivation of spores with the initial inocula of 7.13 and 5.72-log in PBS and apple juice, respectively. And the cell structure of the spores was severely disrupted leading to the leakage of the entocyte. Moreover, the surface properties of the spores were altered, making the adhesion of the spores to the hydrophobic surfaces and the stability of the bacterial suspension decreased leading to more agglomeration of the spores. Further results confirmed the intracellular homeostasis of the spores was also disrupted. Therefore, the inactivation mechanism of DBDP on spores was revealed from different perspectives, and our discoveries promote the theoretical progress in the use of plasma in food sterilization.Industrial relevanceAs an emerging nonthermal preservation technology, cold plasma has been used in food processing. In this study, we confirmed the feasibility of the inactivation of Alicyclobacillus contaminans spores in apple juice by dielectric barrier discharge plasma (DBDP) and investigated the mechanism of plasma inactivation of spores at the molecular biology level. This paper provides preliminary support for the application of low-temperature plasma in food processing such as apple juice.     

Combination of high-frequency ultrasound with propyl gallate for enhancing inactivation of bacteria in water and apple juice

This study evaluated a combination of high-frequency ultrasound (HFU, 1 MHz, 1.6 W/cm²).and a food-grade antioxidant, propyl gallate (PG, 10 mM), to enhance inactivation rates of Listeria innocua and Escherichia coli O157:H7 in water and clarified apple juice. Treatment times ranged from 5 to 20 min. The study also assessed the potential mechanisms of synergistic interactions based on an evaluation of changes in bacterial permeability, morphology, and intracellular oxidative stress. Within 15 min of treatment time, HFU + PG significantly (reduced by 5.5 log CFU/mL, P < 0.05) decreased the bacterial load of both L. innocua and E. coli O157:H7 from an initial inoculum of 6.5 log CFU/mL in both water and clarified apple juice. Overall, L. innocua demonstrated significantly higher resistance to inactivation than E. coli O157:H7 using a combination of HFU + PG. The synergistic antimicrobial activity of HFU+ PG resulted in enhanced membrane damage and oxidative stress induction in bacteria compared to the individual treatments of HFU or PG alone.Industrial relevanceThis study evaluates the synergistic combination of high frequency ultrasound and the food grade antioxidant propyl gallate for non-thermal processing of liquids. The results illustrate significant (>5 log CFU) and rapid inactivation (∼15 min) of inoculated model Gram positive and Gram negative bacteria in apple juice using a synergistic combination of propyl gallate and high frequency ultrasound. The synergistic interactions result in enhanced membrane damage and oxidative stress induction in bacteria. These results illustrate potential of the synergistic non-thermal thermal processing method for processing liquid beverages. Further studies are required for evaluating the scale up and optimization of the novel processing technology and enhancement in quality attributes of beverages.     

Survival of Penicillium expansum and Patulin Production on Stored Apples after Wash Treatments

ABSTRACT: Penicillium expansum is a widespread fungus found on apples that causes fruit decay and may lead to production of a toxic secondary metabolite, patulin. This study was undertaken to evaluate the effectiveness of several chemical sanitizers against P. expansum NRRL 2304 and to establish sanitizing wash treatments that would inhibit P. expansum growth and subsequent patulin production on Empire apples destined for cider. Wash treatments included 200 ppm NaOCl, 1% StorOx®, 0.5% potassium sorbate, 300 ppm SO₂, and 0% to 5% acetic acid. Spores of P. expansum or inoculated apple slices were dipped in sanitizing wash solution for 5 min, and mold growth and patulin production was monitored on subsequent storage. It was found that 0.5% potassium sorbate and 300 ppm SO₂ did not affect mold survival or patulin production; 1% StorOx® was effective against mold spores in solution (4 log Most Probable Number destruction of spores), but there was no significant reduction in spore count when the same solution was used to sanitize mold‐inoculated apple discs. Washing with 200 ppm NaOCl delayed growth of P. expansum on inoculated apple discs but failed to completely inhibit patulin production. Acetic acid solution (2% to 5%) was the most efficient chemical against P. expansum. A wash treatment with ≥2% acetic acid for more than 1 min is recommended to completely inhibit growth of P. expansum and subsequent patulin production on apples destined for cider.     

The potential of atmospheric air cold plasma for control of bacterial contaminants relevant to cereal grain production

The aim of this work was to investigate the efficacy of dielectric barrier discharge atmospheric cold plasma (DBD ACP) against bacteria associated with grains quality and safety. ACP inactivation efficacy was tested against biofilms formed by different strains of E. coli, Bacillus and Lactobacillus in grain model media and against B. atrophaeus endospores either in grain media or attached on abiotic surfaces. Effects were dependent on bacterial strain, media composition and mode of ACP exposure. ACP treatment for 5 min reduced E. coli spp., B. subtilis and Lactobacillus spp. biofilms by > 3 log₁₀, whereas insignificant reductions were achieved for B. atrophaeus. ACP treatment of 5–20 min reduced B. atrophaeus spores in liquids by > 5 log₁₀. Treatment for 30 min reduced spores on hydrophobic surface by > 6 log₁₀, whereas maximum of 4.4 log reductions were achieved with spores attached to hydrophilic surface. Microscopy demonstrated that ACP caused significant damage to spores. In package ACP treatment has potential to inactivate grain contaminants in the form of biofilms, as well as spores and vegetative cells.Industrial relevanceThis study demonstrates that ACP technology is a promising tool for effective bio-decontamination which offers a wide range of possible applications including inactivation of microorganisms on cereal grains. However, due to the nature of the microbial contamination of grains and complex grain structures it may be necessary to optimise the potential for surface inactivation at several stages of grain processing and storage to enhance ACP efficacy against bacterial endospores.     

Inactivation of apple (Malus domestica Borkh) polyphenol oxidases by radio frequency combined with pulsed electric field treatment

Radio frequency (RF) preprocessing combined with pulsed electric field (PEF) processing was employed to inactivate polyphenol oxidase (PPO) in apple juice. PPO enzyme levels, loss of total phenolic content (TPC), colours and volatile components in the apple juice were subsequently determined and compared with conventional processing methods (60 °C for 10 min and 70 °C for 10 min). Results indicated that, when the apple tissue was preprocessed using RF for 10 min, the residual activity of PPO decreased to 13.57%; when the squeezed juice was processed by PEF, the residual activity decreased to about 5% at 15–35 kV cm^?1 for 400 μs. RF treatment caused no significant loss in TPC. Compared with the conventionally processed samples, the apple juice that was RF‐treated for 10 min and PEF‐treated at 15 kV cm^?1 for 400 μs increased its lightness and maintained its fresh‐like flavour.     

Effects of bubble and reactor shape on cold plasma yeast inactivation

Cold plasma is a promising non-thermal technology for inactivating pathogenic and spoilage microorganisms in food by generating various reactive species that damage and inactivate the microorganisms. In this study, a mathematical model was established using COMSOL software to quantitatively describe the process of yeast inactivation by cold plasma under different treatment conditions. The study investigated the effects of discharge voltage, bubble size, and reactor shape on the efficiency of yeast inactivation in apple juice. The results revealed that the discharge voltage significantly influenced yeast inactivation, with a 0.4-fold voltage increase leading to a 53% reduction in inactivation time, achieving a 4-log reduction within 12 min at 21 kV. However, bubble size and reactor shape had no significant impact on the inactivation. The developed model can help to predict the cold plasma inactivation yeast in apple juice, facilitating the design and optimization of plasma-based microbial inactivation in liquid food processing.     

Effect of a combination of electrodialysis with bipolar membranes and mild heat treatment on the browning and opalescence stability of cloudy apple juice

The purpose of this work was to develop a process enabling the quick inactivation of the polyphenol oxidase and pectin methylesterase enzymes, which are present in cloudy or unclarified apple juice; These enzymes are respectively responsible for enzymatic browning and opalescence instability. In order to fulfill this objective, acidification of the apple juice to pH 2.0 was conducted by electrodialysis (bipolar–anionic membranes) followed by mild heat treatment at temperature of 40, 45 and 50 °C for a duration of 0–60 min. Then, juice pH was readjusted to its initial value by electrodialysis with bipolar–anionic membranes. It was shown that a mild heat treatment at 45 °C for 5 min performed on the acidified juice represents an appropriate condition to quickly inactivate the enzymes. Furthermore, the organoleptic properties of the juice after treatment were found to be preserved and the adjusted juice (pH readjusted to its initial value) shows a better color than an untreated apple juice. Opalescence of the adjusted juice was also more stable than for an untreated cloudy apple juice, when stored at 4 °C for 3 months.     

Inactivation of Bacillus cereus spores using a combined treatment of UV-TiO2 photocatalysis and high hydrostatic pressure

Bacillus cereus spores are resistant to high hydrostatic pressure (HHP) processing treatment. A combination of UV-TiO₂ photocatalysis (UVTP for 10 min) and two cycles of 600 MPa HHP treatment for 10 min for the first cycle and 1 min for the second cycle (UVTP-2HHP) at ambient temperature was applied to inactivate B. cereus spores inoculated on a solidified agar matrix (SAM) used as a model matrix. Two cycles of HHP treatment were used as a strategy for induction of spore germination, followed by inactivation. UVTP and 2 cycles of HHP resulted in a 5.0-log CFU/cm² spore reduction (initial spore count was 6.6 log CFU/cm²), including an approximate 0.8-log CFU/cm² reduction due to a synergistic effect. The inactivation mechanism of UVTP pretreatment was related to lipid peroxidation of the spore membrane based on the level of malondialdehyde (MDA) making spores susceptible to the HHP treatment. Flow cytometry and transmission electron microscopic (TEM) analyses showed severe physiological alteration and structural damage to spores after the combined treatment. UVTP and 2 cycles of HHP showed potential for effective inactivation of B. cereus to ensure food safety from B. cereus spores on food products.Practical applicationsInactivation of bacterial spores remains a technical challenge for HHP and other interventions because spores are highly resistant to high pressure. Pretreatment with UVTP followed by two cycles of HHP resulted in reduction in B. cereus spores due to a synergistic effect. This hurdle technology of UVTP and HHP can help food industry in ensuring food safety against the Bacillus spores.     

Some factors governing the production of patulin in apples

The production of patulin by Penicillium patulum and P. expansum growing on apple juice and apple tissue derived from Bramley's seedling apples has been investigated. The carbon sources responsible for the patulin production have been determined using ¹⁴C techniques and growth on defined media.     

Inactivation of Escherichia coli by ozone treatment of apple juice at different pH levels

This research investigated the efficacy of gaseous ozone on the inactivation of Escherichia coli ATCC 25922 and NCTC 12900 strains in apple juice of a range of pH levels, using an ozone bubble column. The pH levels investigated were 3.0, 3.5, 4.0, 4.5 and 5.0. Apple juice inoculated with E. coli strains (10⁶ CFU/mL) was treated with ozone gas at a flow rate of 0.12 L/min and ozone concentration of 0.048 mg/min/mL for up to 18 min. Results show that inactivation kinetics of E. coli by ozone were affected by pH of the juice. The ozone treatment duration required for achieving a 5-log reduction was faster (4 min) at the lowest pH than at the highest pH (18 min) studied. The relationship between time required to achieve 5 log reduction (t_5d) and pH for both strains was described mathematically by two exponential equations. Ozone treatment appears to be an effective process for reducing bacteria in apple juice and the required applied treatment for producing a safe apple juice is dependant on its acidity level.     

Combined high pressure and heat treatment effectively disintegrates spore membranes and inactivates Alicyclobacillus acidoterrestris spores in acidic fruit juice beverage

The commercial potential of high pressure and thermal processing (HPTP) was investigated against Alicyclobacillus acidoterrestris spores in commercial acidic apple juice beverage and in acidified and neutral potassium buffers. With starting spore counts prior to treatments being 6.5 and 7.2 log₁₀ respectively for strains AJA 66 (D_90°C 15.4 min) and ATCC 49025 (D_90°C 8.5 min), HPTP at 600 MPa at 80 °C for 3 min provided an optimal treatment with spore viability reduced below the detection limit for both strains. HPTP at 80 °C for 1 min and HPTP at 70 °C for 3 min achieved 4.1–4.5 log₁₀ CFU/mL reduction. HPTP at 70 °C for 1 min reduced the number of viable spores by 2.0–2.5-log₁₀ CFU/mL. Flow cytometry revealed the presence of membrane-compromised spores among culturable spores following HPTP and heat alone treatments at different temperatures. Electron microscopy clearly showed the efficiency of HPTP with crushed or hollow spores predominating after treatments. No correlation between HPTP susceptibility and genetic diversity was observed for two genotypes of A. acidoterrestris spores. The treatment combination provides a promising option for industrial utility since it requires lower heat and processing time.     

Radio frequency electric fields processing of orange juice

The non-thermal process of radio frequency electric fields (RFEF) has been shown to inactivate bacteria in apple juice at moderately low temperatures, but has yet to be extended to inactivate bacteria in orange juice. An 80 kW RFEF pasteurizer was used to process pulp-free orange juice at flow rates of 1.0 and 1.4 l/min. Escherichia coli K12 in orange juice was exposed to electric field strengths of 15 and 20 kV/cm at frequencies of 21, 30, and 40 kHz. Ascorbic acid (Vitamin C) content and color of the juice before and after treatment were analyzed. Electrical energy costs were calculated using the measured voltage and current. An energy balance was performed using the inlet and outlet temperatures. Processing at an outlet temperature of 65 °C reduced the population of E. coli by 3.3 log relative to the control. Increasing the treatment time and temperature and decreasing the frequency enhanced the level of inactivation. Varying the electric field strength over the range of conditions used had no effect on the inactivation. No loss in ascorbic acid or enzymatic browning was observed due to RFEF processing. The electrical energy determined using the voltage and current was 180 J/ml. This was in good agreement with the energy calculated using the temperature data. The electrical cost was $0.0026/l of orange juice. The results provided the first evidence that the RFEF process inactivates bacteria in orange juice at moderately low temperatures.

Industrial relevance

The RFEF process has been shown to inactivate E. coli in apple juice at moderately low temperatures, but has yet to be extended to inactivate bacteria in orange juice. An RFEF pilot plant pasteurizer was used to process orange juice at rates of up to 1.4 l/min. RFEF processing reduced the population of E. coli by 99.3% at 60 °C and a hold time of 3 s, whereas conventional heating at the same conditions had no effect on the E. coli. This work demonstrated that the non-thermal RFEF process can be extended to inactivate bacteria in orange juice.     

Nonthermal inactivation of polyphenol oxidase in apple juice influenced by moderate electric fields: Effects of periodic on-off and constant exposure electrical treatments

Inactivation of polyphenol oxidase (PPO) in fresh apple juice was studied in moderate electric field (MEF) treatments at field strengths in the range of 85–110 V/cm. The apple juice was subjected to periodic on-off treatments at 50 °C and constant exposure MEF treatments at 50 and 65 °C using two different electro-processing devices, specially constructed for the purpose. PPO inactivation by the electric fields was determined with respect to the corresponding control (nofield) treatments. The application of periodic on-off treatments resulted in about 100% increase (P ≤ 0.05) of the thermal inactivation rate constant of PPO at 50 °C. A similar relative increase of the thermal inactivation rate constant was observed with the constant exposure MEF treatment at 50 °C. The electric field influence was improved significantly (P ≤ 0.05) in the constant exposure MEF treatment at 65 °C. Differences in electrical energy input among the MEF treatments were observed. Results indicate both the electric field strength and the treatment temperature play critical roles in enzyme inactivation by moderate electric fields.Industrial relevanceThis study shows polyphenol oxidase enzyme in fresh apple juice can be significantly inactivated by moderate electric fields applied at below thermal pasteurization temperatures. Therefore, moderate electric field treatment applications are beneficial for the minimally processed fruit and vegetable industry.     

Inactivation of Saccharomyces cerevisiae and Bacillus cereus by pulsed electric fields technology

The effect of pulsed electric field (PEF) treatment, applied in a continuous system, on Saccharomyces cerevisiae and Bacillus cereus cells and spores was investigated. S. cerevisiae inoculated into sterilised apple juice and B. cereus cells and spores inoculated into sterilised 0.15% NaCl were treated with electric field strengths of 10–28 kV/cm using an 8.3 pulse number and with pulse numbers of 4.2–10.4 at 20 kV/cm, respectively. The inactivation of S. cerevisiae depended on the electric field intensity and number of pulses. The yeast inactivation increased when the applied electric field intensity and pulse number were increased. Approximately four log cycles reduction was achieved in apple juice using 10.4 pulses at 20 kV/cm. B. cereus cells were less sensitive to PEF treatment. The reduction in microbial count of B. cereus cells was hardly more than one log cycle using 10.4 pulses at 20 kV/cm. The applied PEF treatment was ineffective on Bacillus cereus spores.     

Inactivation strategy for Clostridium perfringens spores adhered to food contact surfaces

The contamination of enterotoxigenic Clostridium perfringens spores on food contact surfaces posses a serious concern to food industry due to their high resistance to various preservation methods typically applied to control foodborne pathogens. In this study, we aimed to develop an strategy to inactivate C. perfringens spores on stainless steel (SS) surfaces by inducing spore germination and killing of germinated spores with commonly used disinfectants. The mixture of l-Asparagine and KCl (AK) induced maximum spore germination for all tested C. perfringens food poisoning (FP) and non-foodborne (NFB) isolates. Incubation temperature had a major impact on C. perfringens spore germination, with 40 °C induced higher germination than room temperature (RT) (20 ± 2 °C). In spore suspension, the implementation of AK-induced germination step prior to treatment with disinfectants significantly (p < 0.05) enhanced the inactivation of spores of FP strain SM101. However, under similar conditions, no significant spore inactivation was observed with NFB strain NB16. Interestingly, while the spores of FP isolates were able to germinate with AK upon their adhesion to SS chips, no significant germination was observed with spores of NFB isolates. Consequently, the incorporation of AK-induced germination step prior to decontamination of SS chips with disinfectants significantly (p < 0.05) inactivated the spores of FP isolates. Collectively, our current results showed that triggering spore germination considerably increased sporicidal activity of the commonly used disinfectants against C. perfringens FP spores attached to SS chips. These findings should help in developing an effective strategy to inactivate C. perfringens spores adhered to food contact surfaces.     

Inhibitory effect and mechanism of curcumin-based photodynamic inactivation on patulin secretion by Penicillium expansum

Patulin (PAT) contamination caused by Penicillium expansum growth in postharvest fruit brings safety problems. This study investigated the inhibitory effect and mechanism of curcumin (CUR)-based photodynamic inactivation (PDI) on PAT secreted by P. expansum in vitro using a multifunctional light source instrument. After being treated with photosensitive CUR, the growth of mycelium was completely inhibited on potato dextrose agar medium, and the maximum PAT reduction of 93.06% can be achieved (330.0 J/cm², 300 μM CUR) in liquid medium. The inherent mechanism was that PDI causes the reactive oxygen species accumulation in cells, which destroyed the antioxidant defense system and induced severe oxidative damage, resulting in the loss of cell integrity. Furthermore, mycelial apoptosis caused up-regulation of five genes, including PatB, PatE, PatG, PatI, and PatL, but down-regulated ten genes in the PAT biosynthetic pathway. These findings suggest that CUR-based PDI can inhibit fungal infection and toxin secretion.Industrial relevanceThis study found that photosensitized curcumin can significantly inhibit mycelial growth and the secretion of patulin by Penicillium expansum. Curcumin-based photodynamic inactivation has the feasibility of the application on fresh fruits and vegetables, such as apples or oranges, and can avoided excessive waste of resources in picking, transporting, storing, and selling.     

Inactivation kinetics of apple polyphenol oxidase in different pressure–temperature domains

The impact of high hydrostatic pressure and temperature on the stability of polyphenol oxidase (PPO) was studied in cloudy apple juice. Application of 200–500 MPa near room temperature or heat treatment at 45–55 °C at ambient pressure caused an increase of PPO activity of up to 65% in freshly squeezed apple juice. Combined pressure–temperature inactivation experiments with fully activated PPO (5 min treatment at 400 MPa and 20 °C) were carried out in the range of 0.1–700 MPa and 20–80 °C. Enzyme inactivation kinetics followed a 2.2 order reaction scheme at all pressure–temperature conditions tested. A polynomial model was successfully applied to describe the rate of PPO inactivation as a function of pressure and temperature and was used to construct a pressure–temperature isokinetic diagram. This diagram clearly showed synergistic effects of pressure and temperature on the inactivation of apple PPO at pressures above 300 MPa and antagonistic effects at lower pressures. Compared to ambient pressure conditions, temperatures required to inactivate PPO in apple juice were increased 10–15 °C at 100–300 MPa.

Industrial relevance

High pressure processing of fresh fruits is gaining popularity in the food industry because of its ability to inactivate microorganisms and some enzymes near room temperature with little impact on flavour or nutritional attributes of the food. However, quantitative data regarding the impact of process parameters on the target reaction are required to economically utilise this technology. This paper provides a mathematical model describing the combined effect of pressure, temperature and treatment time on the inactivation of PPO in cloudy apple juice.     

Inactivation of Bacillus stearothermophilus spores in egg patties by pressure-assisted thermal processing

The use of pressure-assisted thermal processing (PATP) to inactivate bacterial spores in shelf-stable low-acid foods, without diminishing product quality, has received widespread industry interest. Egg patties were inoculated with Bacillus stearothermophilus spores (10⁶ spores/g) and the product was packaged in sterile pouches by heat sealing. Test samples were preheated and then PATP-treated at 105 °C at various pressures and pressure-holding times. Thermal inactivation of spores was studied at 121 °C using custom-fabricated aluminum tubes; this treatment served as a control. Application of PATP at 700 MPa and 105 °C inactivated B. stearothermophilus spores, suspended in egg matrix rapidly, (4 log reductions in 5 min) when compared to thermal treatment at 121 °C (1.5 log reduction in 15 min). Spore inactivation by PATP progressed rapidly (3 log reductions at 700 MPa and 105 °C) during pressure-hold for up to 100 s, but greater holding times (up to 5 min) had comparatively limited effect. When PATP was applied to spores in water suspension or egg patties, D values were not significantly different. While thermal inactivation of spores followed first-order kinetics, PATP inactivation exhibited nonlinear inactivation kinetics. Among the nonlinear models tested, the Weibull model best described PATP inactivation of B. stearothermophilus spores in the egg product.     

Inactivation of food spoilage bacteria and Escherichia coli O157:H7 in phosphate buffer and orange juice using dynamic high pressure

This study aimed to evaluate the potential of dynamic high pressure (DHP) technology to inactivate pathogenic and spoilage microflora in orange juice. Escherichia coli O157:H7 ATCC 35150, Lactobacillus plantarum ATCC 14917, Leuconostoc mesenteroides ATCC 23386 and two orange juice isolates: Saccharomyces cerevisiae and Penicillium ssp. were subjected individually to different DHP treatments. The effectiveness of DHP treatment was first evaluated in phosphate buffered saline (PBS) before application in orange juice samples. The inactivation efficacy of DHP depended on the pressure applied and the number of passes. It was more efficient against Gram-negative strains than Gram-positives. Complete inactivation and 5 log reduction of E. coli O157:H7 were achieved in orange juice at 200 MPa after 5 and 3 passes at 25 °C, respectively. Lower inactivation was obtained with Penicillium ssp. (4 log), S. cerevisiae (2.5 log), L. plantarum (2.3 log) and L. mesenteroides (1.6 log). The gathered results revealed the potential of DHP to inactivate all the tested microorganisms and then, it could constitute a promising alternative technology for cold pasteurization of fruit juices.     

Growth inhibition and inactivation of Alicyclobacillus acidoterrestris endospores in apple juice by hyperbaric storage at ambient temperature

Control of endospores of Alicyclobacillus acidoterrestris in pasteurized apple juice using hyperbaric storage at 18 to 23 °C was compared to storage at atmospheric pressure and 18 to 23 °C, as well as refrigeration at ~4 °C for up to 30 days. The juice samples were inoculated with approximately 1 × 10⁵ CFU/mL spores. The juice spoiled quickly at atmospheric pressure and ambient temperature, while under refrigeration spore levels remained unchanged for 30 days. Hyperbaric storage of inoculated apple juice at 25, 50 and 100 MPa at 18 to 23 °C resulted in spore inactivation at more rapid rates as pressure magnitudes increased, reaching levels below the detection limit of 10 CFU/mL at 50 and 100 MPa. In highly acid foods such as apple juice, hyperbaric storage at pressures ≤100 MPa and ambient temperature was effective in inactivating spores of A. acidoterrestris for periods up to 30 days.These results indicate hyperbaric storage at ambient temperature as a clearly more efficient preservation procedure to control the development of A. acidoterrestris endospores, compared to ambient temperature and refrigerated storage, in highly acidic foods as apple juice.