Potency of Dimethyl Dicarbonate on the Microbial Inhibition Growth Kinetics, and Quality of Passion Fruit (Passiflora edulis) Juice during Refrigerated Storage.

This study aimed to investigate the effectiveness of dimethyl dicarbonate (DMDC) at various concentrations (0-250 ppm) in inhibiting the growth of Escherichia coli TISTR 117 and spoilage microbes in passion fruit juice (PFJ) and its impact on the physicochemical and antioxidant quality of the juice during refrigerated storage. The highest log reduction in the total viable count, yeast/molds and E. coli was attained in PFJ samples with 250 ppm of DMDC (p ≤ 0.05) added. Microbial growth inhibition by DMDC followed the first-order kinetic model with a coefficient of determination (R2) and inhibition constants (k) ranging from 0.98 to 0.99 and 0.022 to 0.042, respectively. DMDC at 0-250 ppm showed an insignificant effect on pH, °Brix, color (L*, a*, b*), ascorbic acid, total phenolic compound (TPC), total flavonoid content, and antioxidant activity (DPPH, FRAP) (p > 0.05). Control (untreated PFJ), DMDC-250 ppm, and pasteurized (15 s at 72 °C) samples were subjected to 27 days of cold storage at 4 °C. A decreasing trend in pH, total soluble solid, ascorbic acid content, DPPH and FRAP values were observed in all the samples during refrigerated storage. However, the DMDC-250 ppm sample showed a better prospect in physicochemical quality changes compared to the pasteurized and untreated control PFJ samples. ΔE values showed marked changes in the control sample than the DMDC-250 ppm and pasteurized samples at 27 days of storage. Additionally, the total viable count and yeast/mold count were augmented during storage, and an estimated shelf-life of the control, DMDC-250 ppm, and pasteurized samples was approximately 3, 24 and 18 days, respectively. In conclusion, DMDC at 250 ppm could ensure microbial safety without affecting the quality attributes of PFJ during 24 days of storage at 4 °C.

In Silico Safety Assessment of Bacillus Isolated from Polish Bee Pollen and Bee Bread as Novel Probiotic Candidates.

Bacillus species isolated from Polish bee pollen (BP) and bee bread (BB) were characterized for in silico probiotic and safety attributes. A probiogenomics approach was used, and in-depth genomic analysis was performed using a wide array of bioinformatics tools to investigate the presence of virulence and antibiotic resistance properties, mobile genetic elements, and secondary metabolites. Functional annotation and Carbohydrate-Active enZYmes (CAZYme) profiling revealed the presence of genes and a repertoire of probiotics properties promoting enzymes. The isolates BB10.1, BP20.15 (isolated from bee bread), and PY2.3 (isolated from bee pollen) genome mining revealed the presence of several genes encoding acid, heat, cold, and other stress tolerance mechanisms, adhesion proteins required to survive and colonize harsh gastrointestinal environments, enzymes involved in the metabolism of dietary molecules, antioxidant activity, and genes associated with the synthesis of vitamins. In addition, genes responsible for the production of biogenic amines (BAs) and D-/L-lactate, hemolytic activity, and other toxic compounds were also analyzed. Pan-genome analyses were performed with 180 Bacillus subtilis and 204 Bacillus velezensis genomes to mine for any novel genes present in the genomes of our isolates. Moreover, all three isolates also consisted of gene clusters encoding secondary metabolites.

Foodborne pathogen inactivation in fruit juices utilizing commercial scale high-pressure processing: Effects of acidulants and pH.

The effects of juice pH, type of acidulant, and post-treatment refrigeration on the high-pressure processing (HPP) inactivation of Escherichia coli O157:H7, Salmonella enterica, and Listeria monocytogenes in acid beverages were evaluated. Inoculated apple, orange, and grape juices (at their original pH and adjusted to pH 4.00, 4.50, and 5.00) were treated at 550 MPa for 1 min at 5 °C. In addition, inoculated model solutions acidified to a pH of 5.00 with acetic, citric, malic, and tartaric acids were treated at 400 MPa for 1 min at 5 °C. The effect of refrigerated storage for 24 h after treatment on pathogen inactivation in both experiments was also assessed. A greater than 5-log reduction of the three pathogens inoculated was achieved in all juices immediately after HPP at the juices' original pH, and of L. monocytogenes under all experimental conditions. Refrigerated storage for 24 h after HPP treatment improved the inactivation of E. coli O157:H7, to >5-log reduction, at pH 4.00 in apple juice and of Salmonella in the three juices at pH 4.00. The type of acidulant did not significantly (p > 0.01) affect E. coli or Salmonella inactivation in acidified model solutions but a greater than 5-log reduction after HPP was only achieved for L. monocytogenes when acetic acid was used. The effectiveness of HPP for pathogen inactivation depended largely on product pH and the target pathogen of concern.

Survival of Foodborne Pathogens in Low and Nonalcoholic Craft Beer.

Breweries and beverage companies have recently been interested in creating innovative beer varieties that deviate from traditional beer styles, with either low-alcohol content <2.5% alcohol by volume (ABV) or the absence of alcohol altogether (<0.5% ABV). Traditional beers (up to 10% ABV) contain numerous intrinsic and extrinsic factors preventing pathogens from proliferation or propagation. Physiochemical properties such as a low pH, presence of ethanol and hop acids, limited oxygen, and specific processing techniques, including wort boiling, pasteurization, filtration, cold storage, and handling, all contribute to microbial stability and safety. The potential change or absence in one or more of these antimicrobial hurdles can render the final product susceptible to pathogen survival and growth. In this study, the effect of pH, storage temperature, and ethanol concentration on the growth or die-off of foodborne pathogens in low and nonalcoholic beers was evaluated. pH and ethanol concentrations were adjusted from their initial values of 3.65 and <0.50% ABV to pHs 4.20, 4.60, and 4.80; and 3.20 ABV, respectively. The samples were inoculated with individual five-strain cocktails of E. coli O157:H7, S. enterica, and L. monocytogenes, then stored at two different temperatures (4 and 14°C) for 63 days. Microbial enumeration was performed using selective agar with incubation at 35°C. Results showed that nonalcoholic beers allowed for pathogen growth and survival, as opposed to the low-alcoholic ones. E. coli O157:H7 and S. enterica grew approximately 2.00 log  at 14°C, but no growth was observed at 4°C. L. monocytogenes was more susceptible and fell at, or below, the detection limit rapidly in all the conditions tested. The results show that storage temperature is critical in preventing the growth of pathogens. pH did not appear to have a significant effect on the survival of pathogens (p < 0.05). This challenge study demonstrates the need for beverage manufacturers to prioritize and maintain food safety plans along with practices specific to low- and nonalcoholic beer manufacturers.

Probiotic potential of Bacillus Isolates from Polish Bee Pollen and Bee Bread.

The main goal of this study was the evaluation of the probiotic potential of 10 Bacillus spp. strains isolated from 5 bee bread and 3 bee pollen samples. The antagonistic interaction with Staphylococcus aureus and Escherichia coli was a primary criterion for the preliminary selection of the isolates. Three out of ten strains-PY2.3 (isolated from pollen), BP20.15 and BB10.1 (both isolated from bee bread)-were found to be possible probiotic strains. All these strains are safe for humans (exhibiting [Formula: see text]-hemolytic activity) and meet all essential requirements for probiotics in terms of viability in the presence of bile salts and acid conditions, hydrophobicity, auto-aggregation, and co-aggregation with the cells of important human pathogenic bacteria. They also assimilate more than 30% of cholesterol after 24 h of incubation. These three isolates are resistant to penicillin but sensitive (or exhibit moderate resistance) to the other nine antibiotics tested herein. On the basis of whole-genome sequencing, BP20.15 and BB10.1 were classified as B. subtilis and PY2.3 as B. velezensis. Moreover, genomic analyses revealed that all these isolates are potential producers of different antimicrobial compounds, including bacteriocins and secondary metabolites. The outcomes of this study have proven that some of the Bacillus strains isolated from bee pollen or bee bread are potential probiotics.

Whole-genome sequencing and antimicrobial potential of bacteria isolated from Polish honey.

The aim of this study was the whole-genome analysis and assessment of the antimicrobial potential of bacterial isolates from honey harvested in one geographical location-the north of Poland. In total, 132 strains were derived from three honey samples, and the antimicrobial activity of CFAM (cell-free after-culture medium) was used as a criterion for strain selection and detailed genomic investigation. Two of the tested isolates (SZA14 and SZA16) were classified as Bacillus paralicheniformis, and one isolate (SZB3) as Bacillus subtilis based on their ANI and phylogenetic analysis relatedness. The isolates SZA14 and SZA16 were harvested from the same honey sample with a nucleotide identity of 98.96%. All three isolates have been found to be potential producers of different antimicrobial compounds. The secondary metabolite genome mining pipeline (antiSMASH) identified 14 gene cluster coding for non-ribosomal peptide synthetases (NRPs), polyketide synthases (PKSs), and ribosomally synthesized and post-translationally modified peptides (RiPPs) that are potential sources of novel antibacterials. The BAGEL4 analysis revealed the presence of nine putative gene clusters of interest in the isolates SZA14 and SZA16 (including the presence of six similar clusters present in both isolates, coding for the production of enterocin Nkr-5-3B, haloduracin-alpha, sonorensin, bottromycin, comX2, and lasso peptide), and four in B. subtilis isolate SZB3 (competence factor, sporulation-killing factor, subtilosin A, and sactipeptides). The outcomes of this study confirm that honey-derived Bacillus spp. strains can be considered potential producers of a broad spectrum of antimicrobial agents. KEY POINTS: • Bacteria of the genus Bacillus are an important component of honey microbiota. • Honey-derived Bacillus spp. strains are potential producers of new antimicrobials.

Microbial Inhibition by UV Radiation Combined with Nisin and Shelf-Life Extension of Tangerine Juice during Refrigerated Storage.

This study evaluated the efficiency of UV radiation doses (4.68-149.76 J/cm2) and nisin (50-200 ppm) and their combination in comparison with thermal pasteurization on the microbial inhibition kinetics and physicochemical properties of tangerine juice. It was noted that UV-149.76 J/cm2 and nisin (NS) at 200 ppm in conjunction exhibited the highest log reduction in spoilage and pathogenic microbes including Escherichia coli, Lactiplantibacillus plantarum, and Saccharomyces cerevisiae, yeast and molds, and total plate count in tangerine juice. Additionally, the first-order kinetic model provides a better fit for spoilage and pathogenic strains compared with the zero-order model (higher coefficient of determination, R2), particularly for E. coli. UV and NS showed insignificant effects (p > 0.05) on pH, TSS, and TA values compared with pasteurization. However, there were notable differences observed in color analysis, total phenolic compound, total flavonoid content, vitamin C, carotenoid content, and antioxidant activity using DPPH and FRAP assays. The optimized UV + NS samples were subjected to refrigerated storage for 21 days. The results revealed that during the entire storage period, the pH values and the TSS values slightly decreased, and the TA values increased in the treated samples. The UV + NS treatment insignificantly impacted the color properties. The total phenolic, total flavonoid, and carotenoid contents, and vitamin C decreased over time for all sample treatments, whereas the antioxidant properties exhibited varying outcomes, compared with an untreated control and pasteurization. Therefore, UV radiation and nisin (UV-149.76 J/cm2 + NS-200 ppm) in combination could serve as a viable alternative to traditional heat pasteurization of fruit juice during cold storage.

Antioxidants and Quality Changes of Thermally Processed Purple Corn (Zea mays L.) Milk Fortified with Low Sucrose Content during Cold Storage.

Purple corn kernels were subjected to boiling and steaming times of 5-15 min to extract purple corn milk (PCM). Pasteurized and unpasteurized PCM samples were investigated for changes in anthocyanins, antioxidants, and physicochemical properties. Anthocyanins, total phenolics, antioxidant activity, color and viscosity values showed promising results in pasteurized PCM samples extracted from kernels steamed for 5 min (PPCM-S5) compared to other samples (p ≤ 0.05). Changes in L*, a* and b* values, total phenolics and DPPH activity were lowered in PPCM-S5 samples with higher retention of anthocyanins compared to the PCM extracted from boiled kernels (p ≤ 0.05). PCM extracted from 5 min steamed kernels fortified with 4% sucrose (PCM5-S4) after pasteurization revealed the lowest changes in color, pH, total soluble solid and viscosity during 12 days of storage at 4 °C compared to the unpasteurized PCM without sucrose and pasteurized PCM fortified with 6% sucrose. Additionally, pasteurized PCM5-S4 samples marked the highest anthocyanins, total phenolics and antioxidant activity during storage. Microbial load was lowest in pasteurized PCM5-S4 samples stored at 4 °C for 12 days. However, coliforms, yeast or mold and Escherichia coli were not present in the thermally processed PCM samples. The highest sensory scores were obtained in PCM5-S4 at day 12 of storage compared to PCM without any treatment. Therefore, pasteurized PCM extracted from 5 min steamed purple corn kernels retained bioactivity along with 4% sucrose fortification resulted in higher sensory acceptability. As a consequence the shelf-life of PCM5-S4 sample was extended up to 12 days at 4 °C.

Ultrasound-Assisted Extraction of Bioactive Compounds from Cocoa Shell and Their Encapsulation in Gum Arabic and Maltodextrin: A Technology to Produce Functional Food Ingredients.

In this study, the extraction of cocoa shell powder (CSP) was optimized, and the optimized extracts were spray-dried for encapsulation purposes. Temperature (45−65 °C), extraction time (30−60 min), and ethanol concentration (60−100%) were the extraction parameters. The response surface methodology analysis revealed that the model was significant (p ≤ 0.05) in interactions between all variables (total phenolic compound, total flavonoid content, and antioxidant activity as measured by 2,2-Diphenyl-1-picrylhydrazyl (DPPH) and ferric reducing antioxidant power (FRAP assays), with a lack of fit test for the model being insignificant (p > 0.05). Temperature (55 °C), time (45 min), and ethanol concentration (60%) were found to be the optimal extraction conditions. For spray-drying encapsulation, some quality metrics (e.g., water solubility, water activity) were insignificant (p > 0.05). The microcapsules were found to be spherical in shape using a scanning electron microscope. Thermogravimetric and differential thermogravimetric measurements of the microcapsules revealed nearly identical results. The gum arabic + maltodextrin microcapsule (GMM) showed potential antibacterial (zone of inhibition: 11.50 mm; lower minimum inhibitory concentration: 1.50 mg/mL) and antioxidant (DPPH: 1063 mM trolox/100g dry wt.) activities (p ≤ 0.05). In conclusion, the microcapsules in this study, particularly GMM, are promising antioxidant and antibacterial agents to be fortified as functional food ingredients for the production of nutraceutical foods with health-promoting properties.

Purification and characterization of antifungal lipopeptide produced by Bacillus velezensis isolated from raw honey.

Raw honey contains a diverse microbiota originating from honeybees, plants, and soil. Some gram-positive bacteria isolated from raw honey are known for their ability to produce secondary metabolites that have the potential to be exploited as antimicrobial agents. Currently, there is a high demand for natural, broad-spectrum, and eco-friendly bio-fungicides in the food industry. Naturally occurring antifungal products from food-isolated bacteria are ideal candidates for agricultural applications. To obtain novel antifungals from natural sources, we isolated bacteria from raw clover and orange blossom honey to evaluate their antifungal-producing potential. Two Bacillus velezensis isolates showed strong antifungal activity against food-isolated fungal strains. Antifungal compound production was optimized by adjusting the growth conditions of these bacterial isolates. Extracellular proteinaceous compounds were purified via ammonium sulfate precipitation, solid phase extraction, and RP-HPLC. Antifungal activity of purified products was confirmed by deferred overlay inhibition assay. Mass spectrometry (MS) was performed to determine the molecular weight of the isolated compounds. Whole genome sequencing (WGS) was conducted to predict secondary metabolite gene clusters encoded by the two antifungal-producing strains. Using MS and WGS data, we determined that the main antifungal compound produced by these two Bacillus velezensis isolates was iturin A, a lipopeptide exhibiting broad spectrum antifungal activity.

Microbiome analysis of raw honey reveals important factors influencing the bacterial and fungal communities.

Raw honeys contain diverse microbial communities. Previous studies have focused on isolating bacteria and fungi that are culturable, while missing a large proportion of the microbial community due to culture-based constraints. This study utilized next-generation sequencing (NGS) to analyze the composition of microorganisms in raw honey; these data can reveal environmental and physicochemical variables that are associated with different microbial communities. To examine the microbial composition (bacteria and fungi) of raw honey and analyze its association with physicochemical properties, four types of honey (monofloral, wildflower, manuka, and feral; n total = 36) were analyzed via amplicon metagenomics. The analyzed honey samples had relatively similar bacterial communities but more distinct and diverse fungal communities. Honey type was determined as a significant factor influencing alpha and beta diversity metrics of bacterial and fungal communities. For the bacterial communities, titratable acidity (TA) was associated with community richness and diversity. For the fungal communities, Brix, TA, and color were associated with community richness, while water activity and color were associated with community diversity. Additionally, important bacterial and fungal amplicon sequence variants (ASVs) that influenced the overall community were identified. Results from this study provide important insights into the microbial communities associated with different types of raw honey, which could improve our understanding of microbial dynamics in beehives, improve honey production, and prevent honeybee disease.

Kinetic study of selected microorganisms and quality attributes during cold storage of mango and passion fruit smoothie subjected to dimethyl dicarbonate.

The objectives of this research were to study the effect of DMDC (0-250 ppm) on quality and shelf life of mango and passion fruit smoothie during cold storage. The correlation between microbial population (total microorganisms, yeast and mold, E. coli and S. aureus) and DMDC concentration using zero-order kinetic and first-order kinetic was also determined. In addition, the effect of DMDC compared with pasteurization (90 °C, 100 s) on quality of mixed mango and passion fruit smoothie during the cold storage (4 °C) was studied. The results showed that microbial inactivation was best-described by first-order kinetic model due to a higher coefficient of determination (R2). In addition, DMDC did not affect the decreasing trend of total soluble solid, color difference (∆E*) and total phenolic compound as compared to control during the cold storage. DMDC also hindered the increasing trend in microbial population and prevented the loss of antioxidant activity (DPPH and FRAP assays) and total flavonoid content and decreased the PPO activity as compared with the control during the cold storage. In summary, DMDC showed the potential to maintain the quality and to extend the shelf life of mango and passion fruit smoothie during cold storage.

Alicyclobacillus mali sp. nov., Alicyclobacillus suci sp. nov. and Alicyclobacillus fructus sp. nov., thermoacidophilic sporeforming bacteria isolated from fruit beverages.

Six thermo-acidophilic, spore-forming strains were isolated from a variety of juice products and were characterized genetically and phenotypically. According to 16S rRNA and rpoB gene phylogenetic analyses and average nucleotide identity comparisons against the species demarcation cutoff at <95 %, these six strains were determined to represent three novel species of Alicyclobacillus. The isolates were designated FSL-W10-0018T, FSL-W10-0037, FSL-W10-0048, VF-FSL-W10-0049T, FSL-W10-0057 and FSL-W10-0059T. All six isolates were Gram-positive, motile, rod shaped, contained menaquinone 7 as the major respiratory quinone and had ω-cyclohexane C17 : 0 as a major fatty acid. They were all able to grow aerobically in a range of acidic and moderate thermal conditions. Only isolates FSL-W10-0048 and VF-FSL-W10-0049T were able to produce guaiacol. The following names are proposed for the three new species: Alicyclobacillus mali sp. nov. (type strain FSL-W10-0018T =DSM 112016T=NCIMB 15266T); Alicyclobacillus suci sp. nov (VF-FSL-W10-0049T=DSM 112017T=NCIMB 15265T); and Alicyclobacillus fructus sp. nov. (FSL-W10-0059T=DSM 112018T=NCIMB 15264T).

Bee Pollen and Bee Bread as a Source of Bacteria Producing Antimicrobials.

The principal objective of the study was the isolation and identification of bacteria that are present in mature bee bread (BB) and dried (ready for selling and consumption) bee pollen (BP). Obtained isolates were screened for their potential to inhibit select human pathogenic bacteria and their ability to produce enzymes of particular industrial importance. Four and five samples of BP and BB, respectively, were used for the study. In total, 81 strains of bacteria were isolated, and 34 (42%) of them exhibited antagonistic interactions with at least one reference strain of pathogenic bacteria, namely Staphylococcus aureus ATCC 25923, Staphylococcus aureus ATCC 29213, Staphylococcus epidermidis 12228, Pseudomonas aeruginosa ATCC 27857, and Escherichia coli ATCC 25922. The sequencing of the 16S rRNA gene revealed that all strains producing antimicrobials belong to the genus Bacillus spp., and among them, five species were identified: B. pumilus (n = 17), B. altitudinis (n = 9), B. licheniformis (n = 4), B. subtilis (n = 2), and B. safensis (n = 1). Furthermore, 69, 54, 39, and 29 of the strains exhibited lipolytic, proteolytic, cellulolytic, and esterolytic activity, respectively. Alpha amylase and beta galactosidase activity were rarely observed, and none of the strains produced laccase. The outcomes of the study revealed that BP and BB can be considered potential sources of bacteria producing antimicrobial agents and/or enzymes of particular industrial importance. Of course, additional research is required to verify this hypothesis, but the results of preliminary studies are promising.

Bee Bread Exhibits Higher Antimicrobial Potential Compared to Bee Pollen.

This study aimed at investigation of the antimicrobial potential of ethanolic extracts of bee bread (BB) and bee pollen (BP) and suspensions of these products in MHB (Mueller Hinton Broth). We covered 30 samples of BP and 19 samples of BB harvested in Polish apiaries. Slightly lower activity was observed against Gram-negative bacteria compared to Gram-positive staphylococci. BB extracts exhibited higher inhibitory potential with minimum inhibitory concentration (MIC) values in the range from 2.5 to 10% (v/v) against Staphylococcus aureus ATCC 25923 and ATCC 29213. Most active BB extracts, namely, BB6, BB11 and BB19, effectively inhibited growth of clinical isolates of S. aureus (n = 9), including MRSA (methicillin resistant Staphylococcus aureus) strains (n = 3) at concentrations ranging from 2.5 to 5.0% (v/v). Minimal bactericidal concentration (MBC) values were in the same range of concentrations; however, a shift from 2.5 to 5.0% (v/v) was observed for some products. The most active BP extracts inhibited the growth of reference strains of S. aureus at a concentration of 5% (v/v). Up to the concentration of 20% (v/v) three and seven BP extracts were not able to inhibit the growth of S. aureus ATCC 29213 and S. aureus ATCC 25923 respectively. The growth of staphylococci was also importantly inhibited in suspensions of the products in MHB. No correlation between phenolic content and antimicrobial activity was observed.

Evaluation of Foodborne Pathogen Die-off in Back-Sweetened Wine and Apple Cider Models.

ABSTRACT: Wine and alcoholic apple cider are commonly back-sweetened with unpasteurized juice to produce fresh, natural, and palatable sweetened alcoholic beverages. Foodborne pathogens may be introduced from unpasteurized juice into alcoholic beverages through this back-sweetening process. Although foodborne pathogens generally do not survive under low pH conditions or a high alcohol environment, the die-off of these pathogens has not been established to ensure the microbiological safety of the products. To establish the holding conditions that would provide the required 5-log pathogen reduction requirements for these back-sweetened beverages, we evaluated the survival of three common foodborne pathogens, E. coli O157:H7, Salmonella enterica, and Listeria monocytogenes, in modified white grape juice and apple juice models. White grape juice and apple juice were modified with hydrochloric acid and sodium hydroxide and with ethanol to achieve conditions that are similar to back-sweetened white wine and alcoholic apple cider in regard to pH and ethanol content. Foodborne pathogen cocktails were inoculated separately into modified juice models, and their survival in the juice models was recorded over a 96-h period. Our results show that a combination of low pH and high ethanol content resulted in faster pathogen die-off compared with higher pH and lower ethanol conditions. The holding times required for different combinations of pH and ethanol concentration for each juice model to achieve a 5-log reduction were reported. This research provides data to validate pathogen die-off to comply with juice hazard analysis and critical control point 5-log pathogen inactivation requirements for back-sweetened wine and alcoholic apple cider.

Evaluation of high pressure processing (HPP) inactivation of Escherichia coli O157:H7, Salmonella enterica, and Listeria monocytogenes in acid and acidified juices and beverages.

Increasing consumer demand for high-quality foods has driven adoption by the food industry of non-thermal technologies such as high pressure processing (HPP). The technology is employed as a post-packaging treatment step for inactivation of vegetative microorganisms. In order to evaluate HPP inactivation of Escherichia coli O157:H7, Salmonella enterica, and Listeria monocytogenes in acid and acidified juices and beverages, pressure tolerance parameters were determined using log-linear and Weibull models in pH-adjusted apple juice (pH 4.5) at 5 °C. A commercial processing HPP unit was used. The Weibull model better described the inactivation kinetics of the three tested pathogens. According to estimates from the Weibull model, 1.5, 0.9, and 1.5 min are required at 600 MPa to produce 5-log reductions of E. coli, Salmonella, and L. monocytogenes, respectively, whereas according to the log-linear model, 3.2, 1.8, and 2.1 min are required. The effects of process conditions were verified using commercial products (pH between 3.02 and 4.21). In all tested commercial juices or beverages, greater than 5-log reductions were achieved for all tested pathogens using HPP process conditions of 550 MPa for 1 min. These findings demonstrate that the HPP conditions of 600 MPa for 3 min, typically used by the food industry provide an adequate safety margin for control of relevant vegetative pathogens in acid and acidified juices and beverages (pH < 4.5). Results from this study can be used by food processors to support validation studies and may be useful for the future establishment of safe harbors for the HPP industry.

Implementation of ATP and Microbial Indicator Testing for Hygiene Monitoring in a Tofu Production Facility Improves Product Quality and Hygienic Conditions of Food Contact Surfaces: A Case Study.

Rapid ATP testing and microbiological enumeration are two common methods to monitor the effectiveness of cleaning and sanitation in the food industry. In this study, ATP testing and microbiological enumeration were implemented at a tofu production facility with the goal of improving cleaning practices and overall plant hygiene. Results from ATP monitoring were used to target areas of the production environment needing additional cleaning; ATP results were verified by microbiological enumeration of aerobic microorganisms, lactic acid bacteria, and yeasts and molds. Products from the production line were enumerated for the same microorganisms to determine if there was an impact on product quality. After the implementation of ATP monitoring and targeted cleaning, there was a statistically lower proportion of swabs that failed to meet established sanitary requirements for ATP, aerobic microorganisms, and lactic acid bacteria (p < 0.05), but not for yeasts and molds. ATP swabs and microbiological enumeration agreed on site hygiene 75.1% (72.3-77.7%, 95% CI) of the time. Product data indicated that unpasteurized finished products contained a statistically lower microbial load of the three groups of organisms following implementation of the practices (p < 0.05).ImportanceCleaning and sanitation are critical to maintaining safe and high-quality food production. Monitoring these activities is important to ensure proper execution of procedure and to assure compliance with regulatory guidelines. The results from monitoring activities can direct targeted cleaning of areas with higher risk of contamination from foodstuffs and microorganisms. The results of this study show that ATP monitoring and microbiological enumeration are useful tools to verify and improve the efficacy of cleaning and sanitation practices, which can have a positive impact on both plant hygiene and product quality. However, testing regimes and critical parameters will vary based on the product and facility.

Effect of Ethanol Extracts of Propolis (EEPs) against Staphylococcal Biofilm-Microscopic Studies.

Staphylococci growing in the form of biofilm exhibit high resistance to a plethora of antibiotics. The aim of the study was to assess the influence of ethanolic extract of propolis (EEPs) on S. epidermidis ATCC 35984 biofilm using fluorescent microscopy. Propidium iodide (PI) and SYTO 9 were used for differentiation of live and dead cells, and calcofluor white was used to stain the extracellular matrix, the self-produced extracellular polymeric substances (EPS). The outcomes of the research confirm the promising potential of EEPs for eradication of staphylococcal biofilm. However, its activity cannot be classified as fully satisfactory, either in terms of the effectiveness of elimination of bacterial cells or disturbing the EPS structure. A two or even four times higher concentration of EEPs compared to MIC (Minimum Inhibitory Concentration) against planktonic cells (128 µg/mL) was necessary for effective (estimated for 90%) elimination of living cells from the biofilm structure. Unfortunately, even at that concentration of EEPs, the extracellular matrix was only partially disturbed and effectively protected the residual population of living cells of S. epidermidis ATCC 35984. In our opinion, a combination of EEPs with agents disrupting components of EPS, e.g., proteases, lysines, or enzymes degrading extracellular DNA or PIA (polysaccharide intercellular adhesin).

Draft genome sequence of antimicrobial producing Paenibacillus alvei strain MP1 reveals putative novel antimicrobials.

OBJECTIVE: A Paenibacillus strain isolated in previous research exhibited antimicrobial activity against relevant human pathogens including Staphylococcus aureus and Listeria monocytogenes. In this study, the genome of the aforementioned strain, designated as MP1, was shotgun sequenced. The draft genome of strain MP1 was subject to multiple genomic analyses to taxonomically characterize it and identify the genes potentially responsible for its antimicrobial activity. RESULTS: Here we report the draft genome sequence of an antimicrobial producing Paenibacillus strain, MP1. Average Nucleotide Identity (ANI) analysis established strain MP1 as a new strain of the previously characterized Paenibacillus alvei. The genomic analysis identified several putative secondary metabolite clusters including seven Nonribosomal Peptide Synthetase clusters (NRPS) (> 10,000 nt), one bacteriocin or other unspecified Ribosomally Synthesized and Post-Translationally modified Peptide Product (RiPP), one lanthipeptide, and six hybrid clusters (NRPS-Type I Polyketide synthase (T1PKS) and NRPS-trans Amino Transferase Polyketide Synthase (AT-PKS)).