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Microorganisms in processing environments significantly impact the quality and safety of food products and can serve as potential reservoirs for antibiotic-resistant genes, contributing to public health concerns about antimicrobial resistance (AMR). Fish processing plants represent an understudied environment for microbiome mapping. This study investigated the microbial composition, prevalence of Listeria spp., and resistome structures in three catfish processing facilities in the southeastern United States. The 16S rRNA gene sequencing revealed that the observed richness and Shannon diversity index increased significantly from fish to fillet. Beta diversity analysis showed distinct clustering of microbial communities between fish, environment, and fillet samples. Fast expectation-maximization microbial source tracking (FEAST) algorithm demonstrated that the microbiota presents in the processing environment contributed 48.2 %, 62.4 %, and 53.7 % to the microbiota present on fillet in Facility 1 (F1), F2, and F3, respectively. Food contact surfaces made larger contributions compared to the non-food contact surfaces. The linear discriminant analysis of effect size (LEfSe) identified specific microbial genera (e.g., Plesiomohas, Brochothrix , Chryseobacterium and Cetobacterium) that significantly varied between Listeria spp. positive and negative samples in all three processing plants. The metagenomic sequencing results identified 212 antimicrobial resistance genes (ARGs) belonging to 72 groups from the raw fish and fish fillet samples collected from three processing plants. Although there was a significant decrease in the overall diversity of ARGs from fish to fillet samples, the total abundance of ARGs did not change significantly (P > 0.05). ARGs associated with resistance to macrolide-lincosamide-streptogramin (MLS), cationic antimicrobial peptides, aminoglycosides, and beta-lactams were found to be enriched in the fillet samples when compared to fish samples. Results of this study highlight the profound impact of processing environment on shaping the microbial populations present on the final fish product and the need for additional strategies to mitigate AMR in fish products. • Microbiota in processing environment contributed up to 62.4 %, of fillet microbiota. • Food contact surfaces (FCS) impact the microbiota in fillets more than non-FCS. • Plesiomohas, and Brochothrix were more abundant in Listeria positive samples. • Chryseobacterium and Cetobacterium were more abundant in Listeria negative samples. • The richness of the AMR increased while the total abundance remained unchanged. [ABSTRACT FROM AUTHOR]

Abstract This study was conducted to detect and identify microbial populations on pig carcasses at different slaughtering stages and on retail pork cuts at 24 h after slaughter as well as to evaluate the intervention efficiency of sprays containing different concentrations (2% and 4%) of lactic acid. The sprays were applied to the carcass surfaces at the end of the slaughter line. Microbial samples were collected from carcass surfaces after bleeding and after eviscerating, and from retail cuts at 24 h after chilling/spraying. The detected microorganisms were identified through using a Microflex identification instrument and 16S rRNA gene sequencing. The diversity of the bacterial genera; Staphylococcus , Salmonella , Shigella , Enterococci , Escherichia , Acinetobacter and Corynebacterium spp. showed counts ranging from 2.70 to 4.91 log 10 cfu/100 cm2 on the carcasses during slaughter. Most of these genera were also detected on the carcasses after 24 h of chilling. Three species (Staphylococcus hyicus , Acinetobacter albensis , and Corynebacterium xerosis) were also found on the retail cuts of non-sprayed carcasses but not on those of the sprayed groups. Significantly greater reductions in all bacterial species were observed on the carcasses and retail cuts that were sprayed with lactic acid, particularly at the 4% level. Thus, spraying with 4% lactic acid may be an effective intervention for controlling bacterial contamination on pig carcasses to improve the microbiological safety of pork meat. Highlights • A diversity of bacterial species was found on pig carcass surfaces during slaughter. • After 24 h chilling, carcasses were found contaminated with a variety bacterial species. • Spray with lactic acid significantly reduced bacterial species on carcasses and cuts. • Three genus (Staphylococcus , Acinetobacter , Corynebacterium) were found on cuts of control carcasses. • Microbiological safety of pork could be controlled by spraying with 4% lactic acid. [ABSTRACT FROM AUTHOR]

Abstract Time-lapse microscopy methods were used to monitor growth, survival and death of Salmonella enterica serotype Agona individual cells on solid laboratory medium (tryptone soy agar) in the presence of various salt concentrations (0.5%, 3.5%, 4.5% and 5.7% NaCl). The results showed a highly heterogeneous behavior. As NaCl concentration increased, the distribution of the first division time was shifted to higher values and became wider. The mean first division time increased from 1.8 h at 0.5% NaCl to 5.48 h, 16.2 h, and 35.9 h at 3.5%, 4.5% and 5.7% NaCl, respectively. The concentration of NaCl in the growth medium also affected the ability of the cells to divide. The percentage of cells able to grow decreased from 88.9% at 0.5% NaCl to 66.5%, 32.8%, and 6.9% at 3.5%, 4.5% and 5.7% NaCl, respectively. In the latter case (5.7% NaCl), 74 cells out of 406 cells tested (18%) died with mean time to death 5.03 h and standard deviation 6.70 h. To investigate the effect of the behavior of individual cells on the dynamics of the whole population, simulation analysis was used. The simulation results showed that the simultaneous growth, survival and death of cells observed under osmotic stress can lead to a total population behavior known as the "Phoenix" phenomenon. The simulation findings were confirmed by validation experiments using both viable counts and time lapse microscopy. The results of the present study show the high heterogeneity of individual cell responses and the complexity in the behavior of microbial populations at conditions approaching the boundaries of growth. Highlights • High heterogeneity of single-cell response at conditions close to growth boundaries • Simultaneous growth, survival and death can occur under osmotic stress. • Growing, surviving and dying subpopulations can lead to Phoenix phenomenon. [ABSTRACT FROM AUTHOR]

The aim of the current study was to assess the ability of a number of chemicals (acetic Acid (AA), citric acid (CA) lactic acid (LA), sodium decanoate (SD) and trisodium phosphate (TSP)) to reduce microbial populations (total viable count, Campylobacter jejuni , Escherichia coli , Salmonella typhimurium and Listeria monocytogenes ) on raw beef using an immersion system. The following concentrations of each chemical were used: 3 & 5% for AA, CA, LA, SD and 10 &12% for TSP. Possible synergistic effects of using combinations of two chemicals sequentially (LA + CA and LA + AA) were also investigated. L*, a* and b* values were measured before and after treatments and ΔE* values were calculated in order to determine any changes in the color of meat due to the use of these chemicals. In general, all chemical treatments resulted in significantly ( p < 0.05) reduced bacterial counts when compared to untreated controls. The greatest reductions were obtained by using LA3%, SD5%, AA5%, LA5% and SD3% for TVC, C. jejuni, E. coli, S . typhimurium and L. monocytogenes , respectively. However, no significant difference in microbial load was observed between the different concentrations of each chemical used ( p > 0.05). The application of combinations of chemical immersion treatments (LA3% + AA3% and LA3% + CA3%) did not result in further significant reductions in microbial populations when compared to single chemical treatments ( P < 0.05). Assessment of color changes in meat following the application of chemical immersion treatments indicated that using AA or CA at either concentration and LA at 5% led to an increase in the ΔE* value of > 3 immediately after treatment and after 24 h storage. The remaining treatments did not result in significant changes to the color of raw beef. [ABSTRACT FROM AUTHOR]

The main aim of this study was to describe the impact of foliar phenylalanine and urea application on grape and must microbial populations. The tool used to perform the ecological study was DGGE conducted with several infusions in non-enriched and enriched liquid media, as well as direct DNA extractions of grapes and musts. A total of 75 microbial species were found in the study. The alpha diversity indices of grape after both foliar nitrogen treatments did not show significant changes in comparison to the control samples, but were modified in some indices in must samples. The phenylalanine must sample was similar to the control, while foliar urea application caused significant changes in microbial diversity and population structure in comparison to the control must. Further research would be necessary to properly predict the impact on winemaking of the effects observed in this study for grape and must microbiota, especially regarding the foliar application of urea. [ABSTRACT FROM AUTHOR]

The aim of the study was to develop and test the applicability of predictive models for shelf-life estimation of ready-to-eat (RTE) fresh cut salads in realistic distribution temperature conditions in the food supply chain. A systematic kinetic study of quality loss of RTE mixed salad (lollo rosso lettuce-40%, lollo verde lettuce-45%, rocket-15%) packed under modified atmospheres (3% O2, 10% CO2, 87% N2) was conducted. Microbial population (total viable count, Pseudomonas spp., lactic acid bacteria), vitamin C, colour and texture were themeasured quality parameters. Kineticmodels for these indiceswere developed to determine the quality loss and calculate product remaining shelf-life (SLR). Storage experiments were conducted at isothermal (2.5-15 ?C) and nonisothermal temperature conditions (Teff = 7.8 ?C defined as the constant temperature that results in the same quality value as the variable temperature distribution) for validation purposes. Pseudomonas dominated spoilage, followed by browning and chemical changes. The end of shelf-life correlated with a Pseudomonas spp. level of 8 log(cfu/g), and 20% loss of the initial vitamin C content. The effect of temperature on these quality parameters was expressed by the Arrhenius equation; activation energy (Ea) value was 69.1 and 122.6 kJ/mol for Pseudomonas spp. growth and vitamin C loss rates, respectively. Shelf-life prediction models were also validated in real cold chain conditions (including the stages of transport to and storage at retail distribution center, transport to and display at 7 retail stores, transport to and storage in domestic refrigerators). The quality level and SLR estimated after 2-3 days of domestic storage (time of consumption) ranged between 1 and 8 days at 4 ?C andwas predicted within satisfactory statistical error by the kinetic models. Teff in the cold chain ranged between 3.7 and 8.3 ?C. Using the validated models, SLR of RTE fresh cut salad can be estimated at any point of the cold chain if the temperature history is known. Shelf-life models of validated applicability can serve as an effective tool for shelf-life assessment and the development of new products in the fresh produce food sector. [ABSTRACT FROM AUTHOR]

Meta-regression models have gained in popularity during the last years as a way to create more generic models for Microbial Risk Assessments that also include variability. However, as with most meta-analyses and empirical models, systematic biases in the data can result in inaccurate models. In this article, we define experimental bias as a type of selection bias due to the practical limitations of microbial inactivation experiments. Conditions with extremely high D- values (i.e. slow inactivation) need very long experimental runs to cause significant reductions. On the other hand, when the D -value is extremely low, not enough data points can be gathered before the microbial population is below the detection limit. Consequently, experimental designs favour conditions within a practical experimental range, introducing a selection bias in the D -values. We demonstrate the impact of experimental bias in meta-regression models using numerical simulations. Models fitted to data with experimental bias overestimated the z- value and underestimated variability. We propose a rapid heuristic method to identify experimental bias in datasets, and we propose truncated regression to mitigate its impact in meta-regression models. Both methods were validated using simulated data. Thereafter the procedures were tested by building a meta-regression model for actual data for the inactivation of Bacillus cereus spores. We concluded that the dataset included experimental bias, and that it would cause an overestimation of the microbial resistance at high temperatures (>120 °C) for classical meta-regression models. This effect was mitigated when the model was built using truncated regression. In conclusion, we demonstrate that experimental bias could potentially result in inaccurate models for predictive microbiology. Therefore, checking for experimental bias should be a routine step in meta-regression modelling, and be included in guidelines on data analysis for meta-regression. • Experimental bias is a type of selection bias related to experimental limitations. • Because of this bias, the z -value is overestimated and variability is underestimated. • We propose methods to detect and mitigate experimental bias. • A check for experimental bias should be a common step in meta-regression modelling. [ABSTRACT FROM AUTHOR]

Variability and uncertainty are important factors for quantitative microbiological risk assessment (QMRA). In this context, variability refers to inherent sources of variation, whereas uncertainty refers to imprecise knowledge or lack of it. In this work we compare three statistical methods to estimate variability in the kinetic parameters of microbial populations: mixed-effect models, multilevel Bayesian models, and a simplified algebraic method previously suggested. We use two case studies that analyse the influence of three levels of variability: (1) between-strain variability (different strains of the same species), (2) within-strain variability (biologically independent reproductions of the same strain) and, at the most nested level, (3) experimental variability (species independent technical lab variability resulting in uncertainty about the population characteristic of interest) on the growth and inactivation of Listeria monocytogenes. We demonstrate that the algebraic method, although relatively easy to use, overestimates the contribution of between-strain and within-strain variability due to the propagation of experimental variability in the nested experimental design. The magnitude of the bias is proportional to the variance of the lower levels and inversely proportional to the number of repetitions. This bias was very relevant in the case study related to growth, whereas for the case study on inactivation the resulting insights in variability were practically independent of the method used. The mixed-effects model and the multilevel Bayesian models calculate unbiased estimates for all levels of variability in all the cases tested. Consequently, we recommend using the algebraic method for initial screenings due to its simplicity. However, to obtain parameter estimates for QMRA, the more complex methods should generally be used to obtain unbiased estimates. • We compare three methods to estimate variability in microbial kinetics from experimental data. • The simplified algebraic method can be used as an initial assessment for variability. • It is also biased, so its results should not be used as input for QMRA models. • Bayesian models grant more precision and flexibility, at the expense of higher complexity. • Mixed-effects models are robust while being easier to implement than Bayesian models. [ABSTRACT FROM AUTHOR]

Sub-lethal injury within a microbial population, due to processing treatments or environmental stress, is often assessed as the difference in the number of cells recovered on non-selective media compared to numbers recovered on a “selective media” containing a predetermined maximum non-inhibitory concentration (MNIC) of a selective agent. However, as knowledge of cell metabolic response to injury, population diversity and dynamics increased, the rationale behind the conventional approach of quantifying sub-lethal injury must be scrutinized further. This study reassessed the methodology used to quantify sub-lethal injury for Saccharomyces cerevisiae cells (≈ 4.75 Log CFU/mL) exposed to either a mild thermal (45 °C for 0, 10 and 20 min) or a mild pulsed electric field treatment (field strengths of 8.0–9.0 kV/cm and energy levels of 8, 14 and 21 kJ/kg). Treated cells were plated onto either Yeast Malt agar (YM) or YM containing NaCl, as a selective agent at 5–15% in 1% increments. The impact of sub-lethal stress due to initial processing, the stress due to selective agents in the plating media, and the subsequent variation of inhibition following the treatments was assessed based on the CFU count (cell numbers). ANOVA and a generalised least squares model indicated significant effects of media, treatments, and their interaction effects ( P < 0.05) on cell numbers. It was shown that the concentration of the selective agent used dictated the extent of sub-lethal injury recorded owing to the interaction effects of the selective component (NaCl) in the recovery media. Our findings highlight a potential common misunderstanding on how culture conditions impact on sub-lethal injury. Interestingly for S. cerevisiae cells the number of cells recovered at different NaCl concentrations in the media appears to provide valuable information about the mode of injury, the comparative efficacy of different processing regimes and the inherent degree of resistance within a population. This approach may provide similar information for other micro-organisms. [ABSTRACT FROM AUTHOR]

In the present review the latest research studies on Kombucha tea are summarized. Special attention has been paid on microbial population, chemical parameters, biocellulose production, and mainly, on the latest evidences of the biological activities of Kombucha tea. Kombucha tea is a fermented sweetened black or green tea which is obtained from a fermentative process driven by a symbiotic culture of yeast, acetic acid bacteria and lactic acid bacteria. In the last years, its consumption has increasingly grown due to its multiple and potential benefits on human health. This fact has motivated a significant increase in the number of research studies that are focused on the biological activities of this beverage. In this context, this review gathers the main studies that have analyzed the different properties of Kombucha tea (as antioxidant, antimicrobial, antidiabetic, antitumoral, anti-inflammatory, antihypertensive, hepatoprotective, hypocholesterolemic, and probiotic activities). It is highlighted that nowadays few human-based evidences are available to prove the beneficial effect of Kombucha tea on humans' health. In conclusion, further work on Kombucha tea is needed since nowadays few information is available on both clinical studies and the characterization of bioactive compounds and their properties. • Kombucha tea has shown multiple promising results in vitro. • Lack of in vivo and clinical studies hinders its consideration as functional food. • Kombucha-derived biocellulose may present multiple applications in the future. • Further work (bioactivity, clinical trials) is needed to confirm health benefits. [ABSTRACT FROM AUTHOR]

To investigate new disposal methods for agricultural waste from tea plantations, the dynamics of chemical composition and microbial populations of tea plant pruning waste during 60 days of ensiling were studied. The results showed that the pH value decreased markedly in the initial stage and then stabilized after Day 21 (approximately 3.9). At the end of ensiling, lactic and acetic acids were enriched in the silage, while most of the characteristic components of tea, including polyphenols, theanine, and caffeine, were well preserved. Microbiological analysis showed that the relative abundance of Lactobacillus increased rapidly from Day 3 (73.2%), reaching 98.6%. In addition, the dynamics of the microbial community during fermentation were related to its chemical composition and fermentation metabolites. In summary, silage can be an effective approach for treating and utilizing agricultural waste from tea plant pruning. [Display omitted] • Ensiling preserves the main bioactive components in the raw material. • Lactobacillus becomes the dominant genus (73.2%) from Day 3. • Pruned branch from tea plants is a promising bioresource for developing silage. [ABSTRACT FROM AUTHOR]

Abstract: The presence of acylated homoserine lactones (AHLs) and autoinducer-2 (AI-2)-like activity was observed in meat stored under various temperatures (0, 5, 10 and 15°C) and packaging (air, modified atmospheres and modified atmospheres with oregano essential oil) conditions, and correlated with the ephemeral spoilage organisms that comprise the microbial community generally associated with this product. Quorum sensing signal molecules were found to be affected by the packaging conditions e.g. temperature and atmosphere used for meat preservation as a consequence of the development of a distinct microbial community. AHL signal molecules were detected at all incubation temperatures in minced beef samples, both stored aerobically and under modified atmospheres, when both pseudomonads and Enterobacteriaceae populations ranged from 107 to 109 CFU/g, but no signal molecules were detected in minced beef stored under modified atmospheres in the presence of volatile compounds of oregano essential oil, where both these groups failed to grow in high numbers. Additionally, no significant AI-2 activity was observed in the tested cell-free meat extracts (CFME), regardless of the indigenous bacterial populations. The presence of N-(β-ketocaproyl)-homoserine lactone was confirmed with TLC analysis of CFME. [Copyright &y& Elsevier]

Microbial contamination is a persistent problem for grain industry. Many studies have shown that radio frequency (RF) heating can effectively reduce pathogens populations in low moisture foods, but there is a lack on the efficacy to decontaminate natural microbiome. The main objectives of this study were to investigate the effects of different RF heating conditions on natural microbial populations and physicochemical properties of buckwheat. In this study, 30 buckwheat samples collected from 10 different Provinces in China were analyzed for their microbial loads, and the samples with the highest microbial populations were used for further study to select the suitable RF heating conditions. The results showed that microbial loads in tested buckwheat kernels were in the range of 3.4–6.2 log CFU/g. Samples from Shanxi (SX-3) had significantly higher microbial counts than other samples. The selected four temperature-time combinations: 75 °C-20 min, 80 °C-10 min, 85 °C-5 min, and 90 °C-0 min of RF heating could reduce microbial counts to <3.0 log CFU/g in buckwheat kernels at 16.5% w.b. moisture content. Furthermore, the reduction populations of the inoculated pathogens (Salmonella Typhimurium , Escherichia coli , Cronobacter sakazakii, and Bacillus cereus) reached 4.0 log CFU/g under the above conditions, and almost 5.0 log CFU/g especially at high temperature-short holding time combinations (85 °C-5 min and 90 °C-0 min). Besides, physicochemical properties evaluation also showed the insignificant color changes and nutrients loss after RF treatment at 90 °C-0 min. Therefore, the RF heating at 90 °C-0 min holds greater potential than the other lower temperature-longer holding time combinations for applications in buckwheat pasteurization. • Effects of RF heating on microbial populations and physicochemical properties of buckwheat were determined. • Samples from Shanxi had significantly higher microbial counts than others. • Selected four temperature-time combinations resulted in required microbial reductions. • Insignificant color changes and nutrient losses in RF treated buckwheat were observed. [ABSTRACT FROM AUTHOR]

Traditional fermented foods are usually produced by spontaneous fermentation with multiple microorganisms. Environmental factors play important roles in microbial succession. However, it is still unclear how the processing parameters regulate the microbiota during fermentation. Here, we reveal the effects of processing parameters on the core microbiota in spontaneous fermentation of Chinese liquor starter. Rhizopus , Pichia , Wickerhamomyces , Saccharomycopsis , Aspergillus and Saccharomyces were identified as core microbiota using amplicon sequencing and metaproteomics analysis. Fermentation moisture gradually decreased from 34.8% to 14.2%, and fermentation temperature varied between 17.0 °C and 35.3 °C during the fermentation. Mantel test showed that fermentation moisture (P < 0.001) and fermentation temperature (P < 0.05) significantly affected the core microbiota. Moreover, structural equation modelling analysis indicated that fermentation moisture (P < 0.001) and fermentation temperature (P < 0.001) were respectively influenced by the processing parameters, room humidity and room temperature. The succession of Rhizopus , Pichia , Wickerhamomyces , Saccharomycopsis and Aspergillus were significantly affected by room humidity (P < 0.05), and the succession of Saccharomyces was significantly affected by room temperature (P < 0.001). Further, models were constructed to predict the population of core microbiota by room humidity and room temperature, using Gaussian process regression and linear regression (P < 0.05). This work would be beneficial for regulating microorganisms via controlling processing parameters in spontaneous food fermentations. • Humidity and temperature were important processing parameters. • Processing parameters control the core microbiota. • Microbial population can be predicted by models through processing parameters. • The models were beneficial for controlling microbiota in food fermentation. [ABSTRACT FROM AUTHOR]

Abstract: The objectives of this study were to evaluate the effect of different controlled atmospheres, containing 3% O2 +10% CO2, 5% O2 +10% CO2 and 8% O2 +10% CO2, on changes in microbial population of ‘Ambrunés’ sweet cherries throughout storage during 30days and subsequent shelf-life, as well as to identify the main genera of yeast, mould, lactic acid bacteria, Staphylococcus spp., Pseudomonas spp., Enterobacteriaceae spp., and coliforms. The results indicated that controlled atmospheres with 5% O2 +10% CO2 and 8% O2 +10% CO2 were highly effective to control the growth of mesophilic aerobic bacteria, psychrotrophs, Pseudomonas spp., yeasts, and moulds after 15days of storage, showing, in the case of yeasts and moulds, counts that ranged between <1 and 1.75logCFU/g. The genera of yeasts, moulds, lactic acid bacteria, Enterobacteriaceae spp., and coliforms identified were Aureobasidium spp., Penicillium spp., Leuconostoc spp., and Rahnella spp., respectively. In addition, the genera Staphylococcus spp. and Pseudomonas spp. were also identified. On the other hand, cherries of Stage 3 ripening presented the highest counts for all microbial groups. [Copyright &y& Elsevier]

Abstract: Listeria monocytogenes was recently found to enter a long-term-survival (LTS) phase, which may help explain its persistence in natural environments and within food processing plants. The purpose of this study was to investigate the effects of initial cell density, initial pH and type of broth (fresh vs. spent) on the transition of L. monocytogenes to the LTS phase and model the change in viable population density with time. Initial cell density (~106–~1010 CFU/ml) and initial pH (5.36–6.85) both significantly affected the transition of L. monocytogenes to the LTS phase (P <0.001) with initial cell density being the main determining factor. In contrast, type of broth did not significantly affect cell density change during the transition of stationary-phase cells at high initial density to the LTS phase (P >0.05). After 30-d incubation no significant differences in cell densities were observed between either type of broth or between any of the initial cell density/pH treatment combinations (P >0.05), where the mean viable cell density was 4.3±1.1×108 CFU/ml. L. monocytogenes responded to viable cell density in accordance with the logistic equation during transition to the LTS phase. The Agr quorum-sensing system does not appear to play a role in the transition to the LTS phase. Further research is needed to better understand the control mechanisms utilized by L. monocytogenes as it transitions to a coccoid, resistant and stable density state in the LTS phase. [Copyright &y& Elsevier]

Abstract: Interest in potential food applications of ozone has expanded in recent years in response to consumer demands for green technologies. This study was conducted to evaluate the efficacy of gaseous ozone for the microbial reduction and elimination of Salmonella on dried oregano. Ozone treatment was performed up to 120min under continuous stream of two different constant ozone concentrations (2.8 and 5.3mg/L). Significant (P <0.05) reductions of 2.7 and 1.8 log were observed in aerobic plate counts and yeast and mold counts after ozonation at 2.8mg/L for 120min, respectively. Ozonation performed at 5.3mg/L for 90min yielded a reduction of over 3.2 log in the aerobic plate counts. Initial population of a cocktail of Salmonella serotypes (S. Typhimurium, S. Newport and S. Montevideo) on inoculated oregano determined as 5.8logCFU/g decreased significantly by 2.8 and 3.7 log after ozonation at 2.8 and 5.3mg/L for 120min, respectively. Sensory evaluation results suggested that over the 2 log reduction in the microbial population can be obtained on dried oregano by gaseous ozone treatments with an acceptable taste, flavor and appearance. The results demonstrated that the gaseous ozone treatment is an effective alternative microbial reduction technique for dried oregano. [Copyright &y& Elsevier]

Abstract: Spontaneous cocoa bean fermentations carried out in a novel-design 40-kg-capacity stainless steel tank (SST) was studied in parallel to traditional Brazilian methods of fermentation in wooden boxes (40-kg-capacity wooden boxes (WB1) and 600-kg-capacity wooden boxes (WB2)) using a multiphasic approach that entailed culture-dependent and -independent microbiological analyses of fermenting cocoa bean pulp samples and target metabolite analyses of both cocoa pulp and cotyledons. Both microbiological approaches revealed that the dominant species of major physiological roles were the same for fermentations in SST, relative to boxes. These species consisted of Saccharomyces cerevisiae and Hanseniaspora sp. in the yeast group; Lactobacillus fermentum and L. plantarum in the lactic acid bacteria (LAB) group; Acetobacter tropicalis belonging to the acetic acid bacteria (AAB) group; and Bacillus subtilis in the Bacillaceae family. A greater diversity of bacteria and non-Saccharomyces yeasts was observed in box fermentations. Additionally, a potentially novel AAB belonging to the genus Asaia was isolated during fermentation in WB1. Cluster analysis of the rRNA genes-PCR-DGGE profiles revealed a more complex picture of the box samples, indicating that bacterial and yeast ecology were fermentation-specific processes (wooden boxes vs. SST). The profile of carbohydrate consumption and fermentation products in the pulp and beans showed similar trends during both fermentation processes. However, the yeast-AAB-mediated conversion of carbohydrates into ethanol, and subsequent conversion of ethanol into acetic acid, was achieved with greater efficiency in SST, while temperatures were generally higher during fermentation in wooden boxes. With further refinements, the SST model may be useful in designing novel bioreactors for the optimisation of cocoa fermentation with starter cultures. [Copyright &y& Elsevier]

Abstract: Salmonella enterica strains with rdar (red dry and rough) and saw (smooth and white) morphotypes have previously been associated with tomato outbreaks but the dispersal mechanisms of these morphotypes are still poorly understood. In this study, Salmonella adhesion was distinguished from attachment by comparing different contact periods. Initial adhesion of rdar and saw morphotypes of Salmonella was compared in relation to tomato plants with different leaf trichome densities. Trichome densities were increased or reduced by treatment with jasmonic or salicylic acid, respectively. The overall effect of Salmonella morphotype and trichome density on splash dispersal was assessed in a rain simulator and correlated to cell hydrophobicity and initial adhesion. The presence of the rdar morphotype increased initial adhesion at high trichome densities but not at low trichome densities. Attachment of the rdar strain occurred after 30s contact time regardless of trichome density. Splash dispersal was slightly further for the saw morphotype than the rdar morphotype of S. enterica at all trichome densities. Salmonella cells of both morphotypes survived significantly better on the surface of high trichome density leaflets. [Copyright &y& Elsevier]

Abstract: The objective of this study was to develop a new chestnut-based puree, in order to seasonally adjust the offer and use the surplus of undersized production, providing, at the same time, a response to the growing demand for healthy and environmentally friendly products. Broken dried chestnuts have been employed to prepare purees to be fermented with six different strains of Lactobacillus (Lb.) rhamnosus and Lactobacillus casei. The fermented purees were characterized by a technological and sensorial point of view, while the employed strains were tested for their probiotic potential. Conventional in vitro tests have indicated the six lactobacilli strains as promising probiotic candidates; moreover, being the strains able to grow and to survive in chestnut puree at a population level higher than 8 log10 CFU/mL along 40days of storage at 4°C, the bases for the production of a new food, lactose-free and with reduced fat content, have been laid. [Copyright &y& Elsevier]

Abstract: An antimicrobial polyethylene (PE) film was obtained by coating a nisin-based antimicrobial solution. PE sheets were coated on both sides and were used for the packaging of beefsteaks to be stored in air or modified atmosphere packaging (MAP, 60% O2–40% CO2). Microbial populations, species diversity, headspace volatile organic compounds, colour and sensory properties were monitored after 0, 1, 7 and 12days of storage at 4°C. The viable counts showed that there was an effect of MAP and antimicrobial film on the development of all the spoilage associated microbial populations. Carnobacterium spp., Brochothrix thermosphacta, Pseudomonas fragi and Rhanella aquatilis were found in most of the samples. C. maltaromaticum was identified in MRS bulk cells from samples stored in air as well as MAP. Quantitative data of headspace-SPME–GC/MS analysis showed that during storage the production of volatile organic compounds (VOCs) was affected by the use of the treated film and the MAP storage. Compounds such as phenylethylalcohol, nonanal, decanal and ethylbutanoate were produced only from 7 to 12day of storage and only in the samples stored in air. In agreement with the microbiological and VOCs data, the meat stored in active packaging scored the best rankings in the sensory evaluation. Principal component analysis of microbial, sensory and instrumental data showed that beefsteaks stored with the combination of MAP and active packaging for 12days at 4°C differed from the other samples that were more associated to high microbial loads, VOCs concentration and meat off odour perception. In conclusion, the antimicrobial sheets in combination with MAP storage at 4°C were effective for the storage of beefsteaks by retarding the growth of spoilage bacteria, determining lower concentration of VOCs and keeping acceptable levels of colour and other sensory parameters for more than 10days. [Copyright &y& Elsevier]

Abstract: Anchovy Engraulis encrasicolus (L.) is a coastal pelagic and euryhaline species that represents the only European species of the family Engraulidae, with a widespread distribution. In Croatia, it is marketed fresh, frozen, salted or marinated and mainly exported to Italy and Spain, however Anisakis sp. larval infection is frequently the reason for border rejection. Since it is known that the prevalence and intensity of Anisakis infection varies with fish species, fishing area and season, the aim of our study was to identify Anisakis sp. parasitizing European anchovy and infer its population dynamic through a 2.5-year period. Larvae were found coiled and encysted on the external wall of intestine (94%) and reproductive organs (6%), rarely in fillets. Prevalence was 76.1% (95% confidence limits 74.51–77.56%), mean abundance 6.59 (bootstrap 95% confidence limits 5.81–7.26) and mean intensity 8.67 (bootstrap 95% confidence limits 7.82–9.35). The partial CO2 mitochondrial DNA sequence of the isolated anisakids confirmed clustering of the anchovy parasite within A. pegreffii sister group. Parasite population structure showed plasticity inferred by fishing ground, sampling year and fish gender and size. Compared to anisakid prevalence/abundance in other fish, the European anchovy in the Adriatic Sea represents a moderately high-infected paratenic host, although in the Mediterranean and Atlantic waters, anchovies have shown strikingly lesser values of prevalence. Since this host represents one of the most attractive Mediterranean fisheries products traditionally consumed without thermal preparation that in any case would not disrupt larval antigenicity and prevent human allergies, and given the high prevalence of the anisakid within the host, it is necessary to include anchovy into more firm risk assessment frames in order to develop measures that will support the safe alimentary production and consumption of seafood. [Copyright &y& Elsevier]

Abstract: In this work, the establishment of polymicrobial communities on the surfaces which come into contact with the brine during Spanish style Gordal cv. green olive fermentation when subjected to spontaneous or controlled processes (inoculated with Lactobacillus pentosus LPCO10 or 128/2) was studied. Scanning electron microscopy showed that L. pentosus and yeast populations were able to form mixed biofilms throughout the fermentation process on both abiotic (glass slide) and biotic (olive skin) surfaces. The biofilm architectures in both supports were completely different: on the glass slides only aggregates of L. pentosus and yeasts without any polymeric matrix surrounding them were found while on the skin of the fruits, true mature biofilms were observed. During fermentation, the lactic acid bacteria (LAB) population on the olives remained similar while that of yeasts increased progressively to reach similar levels at the end of the process (8–9logCFU/cm2). Molecular analysis showed that different populations of L. pentosus and yeasts were the only microbial members of the biofilm formed during fermentation, regardless of inoculation. Hence, the green olive surface provides an appropriate environmental condition for the suitable development and formation of complex biofilms during controlled or natural table olive processing. [Copyright &y& Elsevier]

Abstract: To fit a lognormal distribution to a complex set of microbial data, including detection data (e.g. presence or absence in 25g) and enumeration data (e.g. 30cfu/g), we compared two models: a model called based on data expressed as concentrations (in cfu/g) or censored concentrations (e.g. <10cfu/g, or >1cfu/25g) versus a model called that directly uses raw data (presence/absence in test portions, and plate colony counts). We used these two models to simulated data sets, under standard conditions (limit of detection (LOD)=1cfu/25g; limit of quantification (LOQ)=10cfu/g) and used a maximum likelihood estimation method (directly for the model and via the Expectation–Maximisation (EM) algorithm for the model . The comparison suggests that in most cases estimates provided by the proposed model are similar to those obtained by model accounting for censorship. Nevertheless, in some cases, the proposed model leads to less biased and more precise estimates than model . [Copyright &y& Elsevier]

Abstract: Isothermal microbial survival curves are usually described by either linear or nonlinear time-dependent models, from which non-isothermal survival curves can be generated if the parameters describing the survival kinetics of the microbial population are known. In order to estimate these parameters, an algorithm based on the steepest decent optimization method was developed. The algorithm searches the values of the survival parameters which minimize the sum of the squared differences between the experimental data and the calculated values provided by the model. The difference of the proposed algorithm with a typical optimization technique is that each data point used is not necessarily coming from the same thermal treatment; instead, data from different non-isothermal processes can be used. The developed algorithm was tested by using published non-isothermal survival data of Salmonella. The data showed that the survival curves can be described by the Weibull model, an already accepted and frequently used nonlinear model. Salmonella''s survival parameters were estimated from the end points and all data points, respectively, of three non-isothermal survival curves. The results obtained showed that the number of survival data points must be sufficiently large to obtain true or statistically sound values of the survival parameters. A suitable way to achieve this is to implement the algorithm using all data points of multiple non-isothermal survival curves or a large number of end points of non-isothermal treatments. Mathematically, the developed algorithm should be applicable to any microbial survival kinetics accurately describing the inactivation of the microorganisms because no specific survival kinetics has to be pre-assumed to run the algorithm. [Copyright &y& Elsevier]

Abstract: Maple sap processing and microbial contamination are significant aspects that affect maple syrup quality. In this study, two sample sets from 2005 and 2008 were used to assess the maple syrup quality variation and its relationship to microbial populations, with respect to processing, production site and harvesting period. The abundance of maple sap predominant bacteria (Pseudomonas fluorescens group and two subgroups, Rahnella spp., Janthinobacterium spp., Leuconostoc mesenteroides) and yeast (Mrakia spp., Mrakiella spp.,Guehomyces pullulans) was assessed by quantitative PCR. Maple syrup properties were analyzed by physicochemical and sensorial methods. Results indicate that P. fluorescens, Mrakia spp., Mrakiella spp. G. pullulans and Rahnella spp. are stable contaminants of maple sap, as they were found for every production site throughout the flow period. Multiple factor analysis reports a link between the relative abundance of P. fluorescens group and Mrakia spp. in maple sap with maple and vanilla odor as well as flavor of maple syrup. This evidence supports the contribution of these microorganisms or a consortium of predominant microbial contaminants to the characteristic properties of maple syrup. [Copyright &y& Elsevier]

Abstract: The overall goal of this study was to develop a set of process design principles for low-energy X-ray irradiation of tree nuts. Almonds and walnuts were inoculated with Salmonella Enteritidis PT30 and Salmonella Tennessee, and conditioned to four different water activities (0.23, 0.45, 0.64, and 0.84 a w ). Thereafter, the inoculated/conditioned samples were irradiated to achieve up to a 5-log reduction in Salmonella using a pilot scale low-energy X-ray food irradiator. Greater efficacy (D 10-value: the dose required to eliminate 90% of the microbial population) for inactivating SE PT30 and S. Tennessee was seen on the surface of almonds (0.226–0.431kGy) than on walnuts (0.474–0.930kGy) at all water activities. Also, the efficacy did not change monotonically with water activity. Overall, no significant difference (P >0.05) in sensory characteristics was seen between non-irradiated almonds and those irradiated to achieve a 5 log reduction in Salmonella. However, irradiating walnuts to the dose corresponding to a 5 log reduction caused a perceivable change in flavor. Post-irradiation storage tests revealed that surviving bacterial counts did not change over 120days, regardless of nut type, Salmonella serovar, and a w . Therefore, low-energy X-ray irradiation technology appears to be a promising non-thermal pasteurization strategy for certain types of nuts. [Copyright &y& Elsevier]

Abstract: The biotype 3 group of the human pathogen Vibrio vulnificus emerged in Israel probably as a result of genome hybridization of two bacterial populations. We performed a genomic and phylogenetic study of V. vulnificus strains isolated from the environmental niche from which this group emerged — fish aquaculture in Israel. The genetic relationships and evolutionary aspects of 188 environmental and clinical isolates of the bacterium were studied by genomic typing. Genetic relations were determined based on variation at 12 variable number tandem repeat (VNTR, also termed SSR) loci. Analysis revealed a new cluster, in addition to the main groups of biotype 1& 2 and biotype 3. Similar grouping results were obtained with three different statistical approaches. Isolates forming this new cluster presented unclear biochemical profile nevertheless were not identified as biotype 1 or biotype 3. Further examination of representative strains by multilocus sequence typing (MLST) of 10 housekeeping genes and 5 conserved hypothetical genes supported the identification of this as yet undiscovered phylogroup (phenotypically diverse), termed clade A herein. This new clonal subgroup includes environmental as well as clinical isolates. The results highlight the fish aquaculture environment, and possibly man-made ecological niches as a whole, as a source for the emergence of new pathogenic strains. [Copyright &y& Elsevier]

Abstract: Kimchi fermentation usually relies upon the growth of naturally-occurring various heterofermentative lactic acid bacteria (LAB). This sometimes makes it difficult to produce kimchi with uniform quality. The use of Leuconostoc mesenteroides as a starter has been considered to produce commercial fermented kimchi with uniform and good quality in Korea. In this study, a combination of a barcoded pyrosequencing strategy and a 1H NMR technique was used to investigate the effects of Leu. mesenteroides strain B1 as a starter culture for kimchi fermentation. Baechu (Chinese cabbage) and Chonggak (radish) kimchi with and without Leu. mesenteroides inoculation were prepared, respectively and their characteristics that included pH, cell number, bacterial community, and metabolites were monitored periodically for 40days. Barcoded pyrosequencing analysis showed that the numbers of bacterial operational taxonomic units (OTU) in starter kimchi decreased more quickly than that in non-starter kimchi. Members of the genera Leuconostoc, Lactobacillus, and Weissella were dominant LAB regardless of the kimchi type or starter inoculation. Among the three genera, Leuconostoc was the most abundant, followed by Lactobacillus and Weissella. The use of Leu. mesenteroides as a starter increased the Leuconostoc proportions and decreased the Lactobacillus proportions in both type of kimchi during kimchi fermentation. However, interestingly, the use of the kimchi starter more highly maintained the Weissella proportions of starter kimchi compared to that in the non-starter kimchi until fermentation was complete. Metabolite analysis using the 1H NMR technique showed that both Baechu and Chonggak kimchi with the starter culture began to consume free sugars earlier and produced a little greater amounts of lactic and acetic acids and mannitol. Metabolite analysis demonstrated that kimchi fermentation using Leu. mesenteroides as a starter was completed earlier with more production of kimchi metabolites compared to that not using a starter, which coincided with the decreases in pH and the increases in bacterial cell number. The PCA strategy using all kimchi components including carbohydrates, amino acids, organic acids, and others also showed that starter kimchi fermented faster with more organic acid and mannitol production. In conclusion, the combination of the barcoded pyrosequencing strategy and the 1H NMR technique was used to effectively monitor microbial succession and metabolite production and allowed for a greater understanding of the relationships between the microbial community and metabolite production in kimchi fermentation. [Copyright &y& Elsevier]

Abstract: The impact of the growth of two Gram-negative bacteria, Psychrobacter celer and Hafnia alvei, inoculated at 102 and 106 cfu/g, on the dynamics of a multispecies community as well as on volatile aroma compound production during cheese ripening was investigated. Results showed that P. celer was able to successfully implant itself in cheese, regardless of its inoculation level. However, when it was inoculated at a high level, the bacterial biodiversity was drastically lowered from day 25 to the end of ripening. Overall, the presence of P. celer led to the higher production of volatile aroma compounds such as aldehydes, ketones and sulfur compounds. Regardless of its inoculation level, H. alvei barely affected the growth of the bacterial community and was subdominant at the end of ripening. It influenced total volatile aroma compound production with volatile sulfur compounds being the most abundant. Overall, these two bacteria were able to implant themselves in a cheese community and significantly contributed to the aromatic properties of the cheese. Their role in flavoring and their interactions with the technological microorganisms must be considered during cheese ripening and should be further investigated. [Copyright &y& Elsevier]

Abstract: The “Zona Alta del Río Mendoza” (ZARM) is the major Malbec grape viticulture region of Argentina. The aim of the present study was to explore Saccharomyces cerevisiae biodiversity in ZARM vineyards. Interdelta PCR and RFLP mtDNA molecular markers were applied to differentiate S. cerevisiae strains. The presence of commercial strains on ZARM vineyards was also assessed. Our results reveal a highly diverse, but genetically closely related, S. cerevisiae population (containing more than 190 molecular patterns among 590 S. cerevisiae isolates). According to the S. cerevisiae strain diversity found in vineyards, they were classified as vineyards with high and low polymorphic S. cerevisiae populations. Six vineyards showed a high polymorphic population, with more than 20 different S. cerevisiae molecular patterns. S. cerevisiae populations in these vineyards were diverse and irregularly distributed, with different strains in each vineyard site. Low polymorphic S. cerevisiae population vineyards displayed very low yeast diversity, with only 9 to 10 different S. cerevisiae strains and presence of two commercial strains widely distributed. Population diversity estimators were calculated to determine the population structure of S. cerevisiae in the ZARM vineyards. The obtained values support the hypothesis that the eight sampled subpopulations come indeed from a larger population. [Copyright &y& Elsevier]

Abstract: Aloreña table olives are naturally fermented traditional green olives with a denomination of protection (DOP). The present study focused on Aloreña table olives manufactured by small and medium enterprises (SMEs) from Valle del Guadalhorce (Southern Spain) under three different conditions (cold storage, and ambient temperature fermentations in small vats and in large fermentation tanks). The microbial load of brines during fermentation was studied by plate counting, and the microbial diversity was determined by a culture-independent approach based on PCR-DGGE analysis. The viable microbial populations (total mesophilic counts, yeasts and molds, and lactic acid bacteria — LAB) changed in cell numbers during the course of fermentation. Great differences were also observed between cold, vat and tank fermentations and also from one SME to another. Yeasts seemed to be the predominant populations in cold-fermented olives, while LAB counts increased towards the end of vat and tank fermentations at ambient temperature. According to PCR-DGGE analysis, microbial populations in cold-fermented olives were composed mostly by Gordonia sp./Pseudomonas sp. and Sphingomonas sp./Sphingobium sp./Sphingopyxis sp. together with halophilic archaea (mainly by haloarchaeon/Halosarcina pallida and uncultured archaeon/uncultured haloarchaeon/Halorubrum orientalis) and yeasts (Saccharomyces cerevisiae and Candida cf. apicola). Vat-fermented olives stored at ambient temperature included a more diverse bacterial population: Gordonia sp./Pseudomonas sp., Sphingomonas sp./Sphingobium sp./Sphingopyxis sp. and Thalassomonas agarivorans together with halophilic archaea and yeasts (mainly S. cerevisiae and C. cf. apicola, but also Pichia sp., and Pichia manshurica/Pichia galeiformis). Some LAB were detected towards the end of vat fermentations, including Lactobacillus pentosus/Lactobacillus plantarum and Lactobacillus vaccinostercus/Lactobacillus suebicus. Only the tank fermentation showed a clear predominance of LAB populations (Lactobacillus sp., Lactobacillus paracollinoides, and Pediococcus sp.) together with some halophilic archaea and a more selected yeast population (P. manshurica/P. galeiformis). The present study illustrates the complexity of the microbial populations in naturally-fermented Aloreña table olives. [Copyright &y& Elsevier]

Abstract: The microbiological quality of the five leading brands of Norwegian bottled still waters was investigated. All brands were free for the enteric indicator organisms and named pathogens whose absence is demanded in current quality directives. The relatively nutrient-poor agar R2A revealed large heterogeneous bacterial populations which grew slowly, or not at all, on clinical media specified for use in substrate-utilization approaches to identification. The main approach used for identification was cultivation of microbes on R2A, followed by amplification and partial sequencing of 16S rDNA genes. The identity of the heterotrophic plate count of the brands differed significantly to that found in many other similar studies with respect to the dominating species. The bacterial flora was dominated by beta- and alphaproteobacteria most of which were psychrotolerant. Several brands contained Sphingomonas and large populations of Methylobacterium species which have been associated with a variety of opportunistic infections of immunocompromised hosts. Analysis of the isolated strains'' nutritional capabilities using the Biolog GN2® system, gave in most instances low positive scores, and strain identifications using the system were generally inconclusive. Measures of assimilable organic carbon in the water revealed that some brands contained levels higher than those which have been associated with biological stability and restricted or no growth of heterotrophs in distribution systems. The relationship between assimilable organic carbon and R2A plate counts was significant and moderately positive for bottled waters. Assimilable organic carbon correlated strongly with the survival time of Escherichia coli when introduced into bottles as a contaminant. Those brands having high values (~100μg/L) supported protracted survival, but not growth of E. coli, whereas E. coli quickly became nonculturable in brands with low values. Thus measures of assimilable organic carbon may have a particular value in predicting the survival of this and nutritionally similar species of hygienic relevance. Only small numbers of fungi were found. However, one isolate (Aureobasidium pullulans) has been associated with infections of humans. [Copyright &y& Elsevier]

Abstract: This study was to screen the ability of biofilm formation by Campylobacter jejuni strains found in New Zealand, and investigate the biofilm growth of C. jejuni in a controlled mixed-microbial population that includes five different bacteria. The ability of C. jejuni to form a biofilm in monoculture and mixed-microbial populations was measured in a laboratory assay using a microtiter plate screening assay. The optical density of the biofilm and cell growth from mixed-microbial populations was converted to a Biofilm Formation Index (BFI). This index was used to standardize the biofilm formation in the mixed-microbial populations. High BFI was observed for Enterococcus faecalis (2.30) and Staphylococcus simulans (3.75) when they were grown with C. jejuni multilocus sequence type ST-474: a dominant poultry and human-associated type in New Zealand. C. jejuni cells were recovered from most of the biofilms containing E. faecalis and/or S. simulans. These results suggest that E. faecalis and S. simulans may play a role in biofilm formation in the poultry environment as both of these microorganisms are found in poultry processing environments and were able to form a biofilm in association with C. jejuni under microaerobic conditions. Understanding the relationships among C. jejuni, E. faecalis and S. simulans in poultry processing plants and farms may help in the design of strategies to reduce the reservoir of contamination of these bacteria and reduce the incidence of campylobacteriosis. [ABSTRACT FROM AUTHOR]

Abstract: Kefir is obtained by milk fermentation with a complex microbial population included in a matrix of polysaccharide and proteins. Several health-promoting activities has been attributed to kefir consumption. The aim of this study was to select microorganisms from kefir able to down-regulate intestinal epithelial innate response and further characterize this activity. Caco-2 cells stably transfected with a human CCL20 promoter luciferase reporter were used to screen a collection of 24 yeast and 23 bacterial strains isolated from kefir. The Toll-like receptor 5 agonist, flagellin was used to activate the reporter cells, while pre-incubation with the selected strains was tested to identify strains with the capacity to inhibit cell activation. In this system, 21 yeast strains from the genera Saccharomyces, Kluyveromyces and Issatchenkia inhibited almost 100% of the flagellin-dependent activation, whereas only some lactobacilli strains showed a partial effect. K. marxianus CIDCA 8154 was selected for further characterization. Inhibitory activity was confirmed at transcriptional level on Caco-2/TC-7 and HT-29 cells upon flagellin stimulation. A similar effect was observed using other pro-inflammatory stimulation such as IL-1β and TNF-α. Pre-incubation with yeasts induced a down-regulation of NF-κB signalling in epithelial cells in vitro, as well as expression of other pro-inflammatory chemokines such as CXCL8 and CXCL2. Furthermore, modulation of CCL20 mRNA expression upon flagellin stimulation was evidenced in vivo, in a mouse ligated intestinal loop model. Results indicate kefir contains microorganisms able to abolish the intestinal epithelial inflammatory response that could explain some of the properties attributed to this fermented milk. [Copyright &y& Elsevier]

Abstract: This study reports the dynamics of microbial populations adhering on the surface of debittered green olives cv. Bella di Cerignola in fermentation sets inoculated with the probiotic strain Lactobacillus paracasei IMPC2.1 in different brining conditions (4% and 8% (w/v) NaCl) at room temperature and 4°C. The probiotic strain successfully colonized the olive surface dominating the natural LAB population and decreasing the pH of brines to ≤5.0 after 30days until the end of fermentation. The dynamics of microbial populations associated with olive surface and belonging to the different groups indicated that inoculated olives held at room temperature did not host Enterobacteriaceae at the end of fermentation. Yeast populations were present in a low number (≤log10 5.7CFU/g) throughout the process. A considerable genetic diversity of LAB species colonizing the olive surface was found mainly in inoculated set brined in 8% NaCl, as indicated by the Shannon diversity index calculated for each set. Generally, strains of Lactobacillus coryniformis, L. paracasei, L. plantarum, L. pentosus, L. rhamnosus, L. brevis, L. mali, L. vaccinostercus, L. casei, Leuconostoc mesenteroides, Leuc. pseudomesenteroides, Lactococcus lactis, Weissella paramesenteroides, W. cibaria, Enterococcus casseliflavus group and E. italicus were identified during the whole process. In particular, L. pentosus was the most frequently isolated species and it showed a high strain diversity throughout fermentation in all processes except for the one held at 4°C. Also a notable incidence of Leuc. mesenteroides on olives was highlighted in this study during all fermentation. Results indicated that the human strain L. paracasei IMPC2.1 can be considered an example of a strain used in the dual role of starter and probiotic culture which allowed the control of fermentation processes and the realization of a final probiotic product with functional appeal. [Copyright &y& Elsevier]

Abstract: The population dynamics of acetic acid bacteria in traditional vinegar production was determined in two independent vinegar plants at both the species and strain level. The effect of barrels made of four different woods upon the population dynamics was also determined. Acetic acid bacteria were isolated on solid media and the species were identified by RFLP-PCR of 16S rRNA genes and confirmed by 16S rRNA gene sequencing, while strains were typed by ERIC-PCR and (GTG)5-rep-PCR. The most widely isolated species was Acetobacter pasteurianus, which accounted for 100% of all the isolates during most of the acetification. Gluconacetobacter europaeus only appeared at any notable level at the end of the process in oak barrels from one vinegar plant. The various A. pasteurianus strains showed a clear succession as the concentration of acetic acid increased. In both vinegar plants the relative dominance of different strains was modified as the concentrations of acetic acid increased, and strain diversity tended to reduce at the end of the process. [Copyright &y& Elsevier]

Abstract: Spoilage bacteria in milk are controlled by treatments such as thermization, microfiltration and addition of carbon dioxide. However, little information is known about the changes in microbial communities during subsequent cold storage of treated milk. Culture-dependent methods and a direct molecular approach combining 16S rRNA gene clone libraries and quantitative PCR (Q-PCR) were applied to obtain a better overview of the structure and the dynamics of milk microbiota. Raw milk samples were treated by the addition of carbon dioxide (CO2), thermization (TH) or microfiltration (MF) and stored at 4°C or 8°C up to 7d. Untreated milk (UT) was used as a control. Psychrotrophic and staphylococci bacteria were enumerated in the milk samples by culture methods. For the molecular approach, DNA was extracted from milk samples and 16S rRNA gene was amplified by PCR with universal primers prior to cloning. The Q-PCR method was used to evaluate the dynamics of dominant bacterial species revealed by clone library analysis of 16S rRNA gene. Comparison of the 16S rRNA gene sequence indicated that the two most abundant operational taxonomic units (OTU), determined at 97% identity, belonged to the class Gammaproteobacteria (40.3% of the 1415 sequences) and Bacilli (40%). Dominant bacterial species in UT, CO2 and TH milk samples at day 3 were affiliated with Staphylococcus, Streptococcus, Clostridia, Aerococcus, Facklamia, Corynebacterium, Acinetobacter and Trichococcus. Dominant bacterial species detected in MF milk were Stenotrophomonas, Pseudomonas and Delftia, while Pseudomonas species dominated the bacterial population of UT, CO2 and MF milk samples at day 7. Staphylococcus and Delftia were the dominant bacterial species in thermized milk. Q-PCR results showed that populations of S. aureus, A. viridans, A. calcoaceticus, C. variabile and S. uberis were stable during 7d of storage at 4°C. Populations of P. fluorescens, S. uberis and total bacteria increased in UT and CO2 milk samples during 7d of storage at 8°C and were noticeable from day 3. This study shows new microbial species which can develop during cold storage after milk treatment and contributes to identifying causes of reduced shelf life and deterioration of technological properties of milk during storage. [Copyright &y& Elsevier]

Abstract: Relatively little is known about the Campylobacter genotypes colonizing extensively reared broiler flocks and their survival through the slaughter process, despite the increasing demand for free-range and organic products by the consumer. Campylobacter isolates from a free-range boiler flock, sampled before and after slaughter, were genotyped by MLST (multilocus sequence typing) and sequence analysis of the flaA short variable region (SVR). The Campylobacter genotypes isolated before and after slaughter were diverse, with up to five sequence types (STs) (seven-locus allelic profiles resulting from MLST) identified per live bird, up to eight STs identified per carcass and 31 STs identified in all. The majority (72.0%) of isolates sampled from carcasses post-slaughter were indistinguishable from those isolated from the live flock before slaughter by ST and flaA SVR type, however, sampling ‘on-farm’ failed to capture all of the diversity seen post-slaughter. There were statistically significant increases in the genetic diversity of Campylobacter (p =0.005) and the proportion of C. coli (p =0.002), with some evidence for differential survival of genotypes contaminating the end product. C. coli genotypes isolated after slaughter were more similar to those from free-range and organic meat products sampled nationally, than from the live flock sampled previously. This study demonstrated the utility of MLST in detecting genetic diversity before and after the slaughter process. [Copyright &y& Elsevier]

Abstract: Classical culturing and denaturing gradient gel electrophoresis (DGGE) techniques have been used for studying the microbial diversity and dynamics of the traditional Spanish Casín cheese during manufacturing and ripening. As with other starter-free cheeses made from raw milk, the microbial diversity of Casín was shown to be high by both culturing and DGGE analyses. The culture technique showed that lactic acid bacteria (LAB) species constituted the majority of the microbial populations. Of the 14 bacterial species identified, Lactococcus garvieae was predominant in the three-day-old cheese sample, although it was replaced by Lactococcus lactis subsp. lactis at day 30. As expected, the DGGE profiles obtained were complex, consisting, depending on the sample, in five to ten different amplification bands. Among these, a band corresponding to Streptococcus thermophilus was observed throughout the whole manufacturing process. This species had never been identified from traditional Spanish cheeses previously. Culturing and molecular methods showed high populations of undesirable microorganisms, arguing for a required improvement in the hygiene of Casín manufacture. Random amplification of polymorphic DNA (RAPD) profiling suggested that the L. garvieae and L. lactis populations were composed of one and five strains, respectively. In addition, only a single L. lactis RAPD pattern was stably maintained from day three to day 30, indicating high succession of strains along ripening. After a thoroughly characterisation, strains of the two Lactococcus species could be used in designing specific starter cultures for Casín. Additional species (such as Lactobacillus plantarum and Corynebacterium variabile) might be included as adjunct cultures. [Copyright &y& Elsevier]

Abstract: A Food Safety Management System (FSMS) implemented in a food processing industry is based on Good Hygienic Practices (GHP), Hazard Analysis Critical Control Point (HACCP) principles and should address both food safety control and assurance activities in order to guarantee food safety. One of the most emerging challenges is to assess the performance of a present FSMS. The objective of this work is to explain the development of a Microbial Assessment Scheme (MAS) as a tool for a systematic analysis of microbial counts in order to assess the current microbial performance of an implemented FSMS. It is assumed that low numbers of microorganisms and small variations in microbial counts indicate an effective FSMS. The MAS is a procedure that defines the identification of critical sampling locations, the selection of microbiological parameters, the assessment of sampling frequency, the selection of sampling method and method of analysis, and finally data processing and interpretation. Based on the MAS assessment, microbial safety level profiles can be derived, indicating which microorganisms and to what extent they contribute to food safety for a specific food processing company. The MAS concept is illustrated with a case study in the pork processing industry, where ready-to-eat meat products are produced (cured, cooked ham and cured, dried bacon). [Copyright &y& Elsevier]

Abstract: It is well known that fresh-cut processors usually rely on wash water sanitizers to reduce microbial counts in order to maintain quality and extend shelf-life of the end product. Water is a useful tool for reducing potential contamination but it can also transfer pathogenic microorganisms. Washing with sanitizers is important in fresh-cut produce hygiene, particularly removing soil and debris, but especially in water disinfection to avoid cross-contamination between clean and contaminated product. Most of the sanitizing solutions induce higher microbial reduction after washing when compared to water washing, but after storage, epiphytic microorganisms grow rapidly, reaching similar levels. In fact, despite the general idea that sanitizers are used to reduce the microbial population on the produce, their main effect is maintaining the microbial quality of the water. The use of potable water instead of water containing chemical disinfection agents for washing fresh-cut vegetables is being advocated in some European countries. However, the problems of using an inadequate sanitizer or even none are considered in this manuscript. The need for a standardized approach to evaluate and compare the efficiency of sanitizing agents is also presented. Most new alternative techniques accentuate the problems with chlorine suggesting that the industry should move away from this traditional disinfection agent. However, the use of chlorine based sanitizers are presented as belonging to the most effective and efficient sanitizers when adequate doses are used. In this review improvements in water disinfection and sanitation strategies, including a shower pre-washing step and a final rinse of the produce, are suggested. [Copyright &y& Elsevier]

Abstract: Predictive microbiology is the area of food microbiology that attempts to forecast the quantitative evolution of microbial populations over time. This is achieved to a great extent through models that include the mechanisms governing population dynamics. Traditionally, the models used in predictive microbiology are whole-system continuous models that describe population dynamics by means of equations applied to extensive or averaged variables of the whole system. Many existing models can be classified by specific criteria. We can distinguish between survival and growth models by seeing whether they tackle mortality or cell duplication. We can distinguish between empirical (phenomenological) models, which mathematically describe specific behaviour, and theoretical (mechanistic) models with a biological basis, which search for the underlying mechanisms driving already observed phenomena. We can also distinguish between primary, secondary and tertiary models, by examining their treatment of the effects of external factors and constraints on the microbial community. Recently, the use of spatially explicit Individual-based Models (IbMs) has spread through predictive microbiology, due to the current technological capacity of performing measurements on single individual cells and thanks to the consolidation of computational modelling. Spatially explicit IbMs are bottom–up approaches to microbial communities that build bridges between the description of micro-organisms at the cell level and macroscopic observations at the population level. They provide greater insight into the mesoscale phenomena that link unicellular and population levels. Every model is built in response to a particular question and with different aims. Even so, in this research we conducted a SWOT (Strength, Weaknesses, Opportunities and Threats) analysis of the different approaches (population continuous modelling and Individual-based Modelling), which we hope will be helpful for current and future researchers. [Copyright &y& Elsevier]

Abstract: We studied the diversity and dynamics of the microbiota of Calenzana, a Corsican raw milk cheese by microbial counting and culture-independent methods (TTGE and DGGE). Cheese from two farms, one producing goat cheese and the other one sheep cheese, was studied. The usual process for cheese making, without starter adjunct, was used. Lactococci and mesophilic lactobacilli were the dominant components of the flora during the early stages of the process. Microbial counting showed that the populations of salt-tolerant bacteria, yeasts and moulds were lower than in other artisanal Corsican cheeses. This difference was probably due to the surface microflora being removed during ripening. TTGE indicated that Lactococcus lactis ssp. lactis was the dominant subspecies throughout the process of Calenzana cheese making. DGGE showed the presence of numerous surface bacteria, (coryneforms) and various Gram-negative bacteria. Relationships between physico-chemical characteristics of the cheese and microflora change were also established. For example, the high NaCl content may explain the decrease of the lactic acid bacterial population during ripening. This study shows the consequences of various technological parameters on the diversity and dynamics of dairy microbial community. [Copyright &y& Elsevier]

Abstract: The combined effects of caprylic acid and mild heat were investigated to ascertain their impact on Cronobacter spp. (E. sakazakii) in reconstituted infant formula. Samples containing a mixture of 3 strains of Cronobacter spp. (107 to 108 CFU/ml) were prepared with various concentrations of caprylic acid (5, 10, 20, and 30 mM) and were then heated to 45, 50, and 55 °C. The inhibitory effect of the combined treatment resulted in a synergistic effect, in which Cronobacter spp. numbers were reduced much more rapidly with increased temperatures and concentrations of caprylic acid. When samples were treated with 30 mM caprylic acid, the time required to reduce Cronobacter spp. cell numbers to an approximate reduction of 7.8 log CFU/ml was 60 min at 45 °C, 20 min at 50 °C, and 10 min at 55 °C. In the validation assay using a low population of Cronobacter spp. (approximately 103 log CFU/ml), no recovery of injured cells was observed after samples were treated with 10 mM caprylic acid for 20 min at 55 °C, 20 mM caprylic acid for 10 min at 50 °C and 55 °C, and 30 mM caprylic acid for 10 min at 45 °C to 55 °C. To determine the bactericidal mechanism of caprylic acid, membrane integrity was examined by fluorescent staining followed by flow cytometry and confocal microscopy. Increased cellular inactivation was associated with increased propidium iodide staining, indicating damage to the cell membrane of Cronobacter spp.. Overall, these data indicate that the addition of this natural antimicrobial agent to infant formula may have potential use for controlling microbes prior to consumption at lower heating temperatures. The study also provides a complementary understanding of the mode of action of caprylic acid on Cronobacter spp. [Copyright &y& Elsevier]

Abstract: In this study we investigated the microbiota of Gorgonzola rinds and maturing shelf swabs collected in 5 different maturing cellars in the Northwest part of Italy, in association with the detection and characterization of Listeria monocytogenes. Culture-dependent and -independent methods were performed in order to profile the main microbial populations present on the rinds and in the maturing shelves and species-specific PCR and Pulsed Field Gel Electrophoresis (PFGE) were used to identify and type L. monocytogenes isolates. The microflora was predominated by lactic acid bacteria and coagulase negative cocci, while enterococci and yeasts were very variable between the samples. Arthrobacter sp., Carnobacterium sp., Staphylococcus sp. and Brevibacterium linens, as bacteria, and Debaryomyces hansenii, as yeast, were detected by Denaturing Gradient Gel Electrophoresis (DGGE). Cluster analysis of the DGGE profiles clearly highlighted a cellar-specific microflora. L. monocytogenes was isolated in 11.1% of the rinds and 29.4% of the swabs and the molecular characterization of the isolates suggests a route of contamination from the maturing shelves to the rinds. No correlation was found between DGGE profiles and presence or absence of L. monocytogenes. [Copyright &y& Elsevier]

Abstract: In this work microbiological shelf life of green tea infusions processed by pulsed electric fields (PEF) treatment (38.4 kV/cm for 200 μs) were assessed at the three storage temperatures of 4, 25 and 37 °C, respectively. Immediately after PEF treatment, no viable bacterial cells were detected. However, significant recovery was observed during storage at 25 and 37 °C. The total aerobic microbial population increased rapidly in PEF-treated green tea infusions when stored at 25 and 37 °C for 14 and 7 days respectively. However, the microbial population remained under 1 log10 CFU/ml when stored at 4 °C up to 180 days. These results demonstrated that a certain proportion of microbial cells in green tea infusions were sublethally injured by PEF treatment, and the recovery of apparently dead or ‘intermediately damaged’ cells could increase the detected number of surviving microorganisms. In the present study PEF-treated green tea infusions were placed at 4 °C for various time, and then stored at 37 °C. Cold storage (4 °C for 7 days) following PEF treatment was found to be effective to delay or inhibit the repair process of sublethally injured cells and extend the microbiological shelf life of PEF-treated green tea infusions to more than 90 days at 37 °C. [Copyright &y& Elsevier]

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. • Kinetic models of microbial inactivation caused by DMDC were determined. • Microbial inactivation best fitted by first-order kinetic model with R2 > 0.9000. • DMDC has a potential to preserve quality attributes of smoothie sample. • DMDC treatment presented a high microbial stability during cold storage at 4 °C. • Shelf life of smoothie sample during cold storage was extended by DMDC. [ABSTRACT FROM AUTHOR]

Abstract: The NaCl growth limits of individual Salmonella enteritidis cells were studied. Six different levels of the pathogen (from 101 to 106 cells) were inoculated on tryptic soy agar plates at NaCl concentrations ranging from 0.5 to 8% and pH values of 5.0, 5.5 or 7.3. The NaCl growth limits of individual cells were estimated from the ratio between the number of colonies formed at each NaCl concentration interval and the number of cells initially inoculated, based on the assumption that each colony was derived from a single cell. The results showed that, as the NaCl concentration increased and the pH decreases, the number of cells able to grow and form a colony gradually decreased indicating variability in the growth limits of individual cells. The distribution of single-cell growth limits was used to predict the behavior of microbial populations. The results showed that as the inoculum size increased the NaCl growth limits of S. enteritidis increased and became less variable. Furthermore, at conditions close to the boundary of growth the presence of a non-growing fraction in the population resulted in a delay of population growth, which we called “pseudo-lag”. The extent of “pseudo-lag” is determined by the environment while its variability is affected by both the inoculum size and the growth conditions. At growth-limiting conditions the total “apparent lag” of the population is a convolution of the “pseudo-lag” and the “physiological lag” of the growing cells. The data on the variability in the growth limits of single microbial cells presented in this work stress the need for stochastic approaches in quantitative microbiology especially at environments close to the boundary of growth. [Copyright &y& Elsevier]

Abstract: Cassava fish (Pseudotolithus sp.) was naturally fermented on three different occasions at room temperature (28–30 °C) for 3 to 8 days and the microbial population and the occurrence of various bacterial species were monitored. In general, after a slight increase during the early stages of fermentation, the microbial population decreased as the fermentation progressed. A total of 224 isolates belonging to the genera Bacillus, Staphylococcus, Micrococcus, Streptococcus, Corynebacterim, Pseudomonas, Achromobacter and Alcaligenes were isolated from the fermenting fish samples and the predominant ones were identified at the species level. Bacillus spp. and Staphylococcus spp. were the predominant genera and accounted for 48.7% and 27.3% of the total number of isolates respectively. Bacillus subtilis and Bacillus licheniformis, Staphylococcus lentus and Staphylococcus xylosus and Micrococcus luteus were the representative strains isolated during the fermentation. Bacillus spp., Staphylococcus spp. and Micrococcus spp. initiated the fermentation but Micrococcus spp. were few in numbers and died off after four (4) days of fermentation while Bacillus spp. and Staphylococcus spp. persisted up to the end of fermentation. Enterobacteriaceae were fewer in numbers at the start of fermentation (102 cfu/g) and their numbers decreased to less than 10 cfu/g after two days of fermentation. Bacillus spp. as well as Staphylococcus spp. isolated possessed moderate proteolytic and lipolytic activities. Micrococcus spp. showed weak proteolytic activity and no lipolytic activity. [Copyright &y& Elsevier]