Using the gamma concept in modelling fungal growth: A case study on brioche-type products.

Fungi are common spoilers of intermediate moisture foods such as bakery products. Brioche are bakery products prone to fungal spoilage due to their pH (5.8-6.2) and water activity (aw) (0.82-0.84). The aims of the present study were: (i) the identification of fungal species occurring in brioche products, (ii) the in vitro assessment of their growth potential, and (iii) the development of a validated growth model following the gamma concept. A total of 102 fungal strains were isolated, with Penicillium sp., Cladosporium sp., and Aspergillus sp. being the main genera, representing 90% of the isolates. Given the isolation frequency, any potential fungal prevalence throughout the bakery processs and/or the results of in vitro assessment of fungal growth potential under conditions mimicking brioche (pH, aw, temperature), Aspergillus flavus, Aspergillus fumigatus, and Penicillium sp. were selected for the development of the gamma model. According to in vitro validation, the model successfully predicted fungal growth, while on in situ experiments, the intrinsic parameters (aw and/or level of used preservative) of brioche in combination with packaging conditions (modified atmosphere) did not allow fungal growth.

A case-control study of hopelessness and suicidal behavior in the city of Athens, Greece. The role of the financial crisis.

This study aimed to examine the psychometric properties of Beck Hopelessness Scale (BHS) in Greek patients with a recent suicide attempt, the predictive ability of BHS especially in terms of age and the possible effect of the financial crisis on hopelessness as it has been identified as a significant suicide risk factor. The study included a total of 510 individuals, citizens of Athens, Greece. Three hundred and forty individuals (170 attempted suicide patients, and 170 -age and gender- matched healthy individuals used as controls) completed BHS before the financial crisis onset (from 2009 to 2010). A sample of 170 healthy individuals also completed the BHS after the financial crisis onset (from 2013 to 2014), when the impact of the austerity measures was being felt by the Greek society. The Greek BHS version demonstrated good psychometric properties and a sufficient degree of internal consistency. Attempted suicide patients (M = 9) presented higher hopelessness than the controls (M = 3). The patients' age positively correlated with hopelessness (rs = .35, p < .001), and the ROC curve revealed that the BHS ability to discriminate the individuals with pathological rates of hopelessness from those without, strengthened as age increased. No significant differences were found between the individuals evaluated before (M = 3) and after (M = 3, p > .05) the crisis onset. Nevertheless, regarding the latter group, the women, the younger individuals, the less educated, the unemployed and the participants with low or mediocre self-reported financial status presented increased hopelessness. Our findings suggest that BHS could be a useful instrument especially when screening for suicide risk in people of older age, and that the financial crisis in Greece had a greater impact on subgroups of the population regarding hopelessness.

Studying the metabolic factors that may impact the growth of co-cultured Listeria monocytogenes strains at low temperature.

The simultaneous presence of more than one strains of Listeria monocytogenes in the same food product may affect the growth capacity of each strain. The present study evaluated the metabolites composition that may potentially influence the growth of individual L. monocytogenes strains in a dual strain composite. Based on previous studies, L. monocytogenes strains, C5 (4b) and 6179 (1/2a) were selected due to the remarkable interaction, which was observed during their co-culture. The selected strains were inoculated (2.0 - 3.0 log CFU/mL) in Tryptic Soy Broth with 0.6% Yeast Extract (TSB-YE) in single and two-strain cultures (1:1 strain ratio). Bacterial growth was assessed during storage at 7 °C, under aerobic conditions (AC). Their resistance to different antibiotics enabled the selective enumeration of each strain in the co-culture. After reaching stationary phase, single and dual cultures were centrifuged and filtered. The cell-free spent medium (CFSM) was either characterized by Fourier transform infrared (FTIR-ATR) spectrometry or re-inoculated, after the addition of concentrated TSB-YE (for nutrient replenishment), with single and two-strain cultures for the evaluation of growth under the influence of metabolites produced from the same singly and co-cultured strains in the different combinations of strains and CFSM origin (7 °C/AC) (n = 2x3). By the end of storage, singly-cultured C5 and 6179 had reached 9.1 log CFU/mL, while in dual culture, 6179 was affected by the presence of C5 attaining only 6.4 ± 0.8 log CFU/mL. FTIR-ATR spectra of CFSM produced by singly-cultured 6179 and the co-culture were almost identical. Characteristic peaks in FTIR-ATR spectrum of CFSM of singly-cultured C5 at 1741, 1645 and 1223 cm-1 represent functional groups which were not present in the CFSM of the co-culture. These molecules may be located intracellularly or mounted on bacterial cell surface and removed from the supernatant during cell filtration of the co-culture. Both singly- and co-cultured 6179 managed to grow similarly regardless of CFSM origin. Contrarily, both singly- and co-cultured C5 managed to outgrow 6179 in CFSM which contained high concentration of C5 metabolites, while in CFSM produced by singly-cultured 6179, C5 did not grow, suggesting that the produced metabolites of strain 6179 appears to be harmful to strain C5. However, during co-culture, C5 may produce molecules that counteract the inhibitory effect of 6179. The findings shed more light on the mechanism behind the inter-strain interactions of L. monocytogenes indicating that both contact of cells and extracellular metabolites may influence the behavior of the different co-existing strains.

Studying the effect of oxygen availability and matrix structure on population density and inter-strain interactions of Listeria monocytogenes in different dairy model systems.

Due to the ubiquitous character of Listeria monocytogenes multiple strains of the pathogen may end up co-existing in/on the same final products and could potentially cause infection during consumption. Such multiple strain contamination may occur in different stages of the food supply chain. The present study evaluated the effect of oxygen availability and matrix structure on inter-strain interactions of L. monocytogenes that may occur at high population levels in/on different dairy model systems. L. monocytogenes strains C5 and ScottA (4b), 6179 (1/2a) and PL25 (1/2b) were selected as resistant to different antibiotics (enabling selective enumeration of each strain in co-culture) and inoculated (2.0-3.0 log CFU/mL, g or cm2) in Ricotta and Camembert broth (1 dairy product: 2 » Ringer solution) and in/on dairy-based structured media (dairy broth supplemented with 0.6 and 1.4% agar), in single and two-strain cultures (1:1 strain ratio). Bacterial growth was assessed during storage at 7 °C, under aerobic, hypoxic and anoxic conditions. Every experimental treatment was tested with three biological replicates and two technical repeats (n = 3 × 2). The simultaneously presence of different strains of the pathogen in/on the same substrate did not affect neither the duration of the lag phase nor the growth rate of the co-cultured strains. The observed inter-strain interactions were related with the final population reached/decrease during storage and occurred after the "critical" population density of ca. 6.0 log CFU/mL, g or cm2. The phenomenon was more pronounced in/on Ricotta than in/on Camembert-based substrates, indicating that the composition and the available nutrients of the substrate may affect the interactions that expressed as difference in the final population level between singly and co-cultured strains. Under aerobic and hypoxic conditions, most of the observed interactions were more pronounced in dairy-based broths and were mitigated with the addition of agar. The elimination of oxygen resulted in a prolonged lag time, which lasted at least 5 days and no observed interactions by the end of storage, due to low microbial counts. Investigating inter-strain interactions during growth in/on different substrates, which may have undergone temperature abuse during their transport along the supply chain or during storage in household refrigerators, could assist in explaining the mismatch between clinical and food samples during outbreak investigations.

Short-Term Effects of Traditional Greek Meals: Lentils with Lupins, Trahana with Tomato Sauce and Halva with Currants and Dried Figs on Postprandial Glycemic Responses-A Randomized Clinical Trial in Healthy Humans.

Low glycemic index (GI) diets have been associated with decreased chronic disease risk. In a randomized, cross-over study we investigated the GI and glycemic response to three traditional Greek mixed meals: Lentils, Trahana, and Halva. Twelve healthy, fasting individuals received isoglucidic test meals (25 g available carbohydrate) and 25 g glucose reference, in random order. GI was calculated and capillary blood glucose (BG) samples were collected at 0-120 min after meal consumption. Subjective appetite ratings were assessed. All three tested meals provided low GI values. Lentils GI was 27 ± 5, Trahana GI was 42 ± 6, and Halva GI was 52 ± 7 on glucose scale. Peak BG values were lowest for Lentils, followed by Trahana and then by Halva (p for all <0.05). Compared to the reference food, BG concentrations were significantly lower for all meals at all time-points (p for all <0.05). Lentils provided lower glucose concentrations at 30 and 45 min compared to Trahana (p for all <0.05) and at 30, 45, and 60 min compared to Halva (p for all <0.05). BG concentrations did not differ between Trahana and Halva at all time points. No differences were observed for fasting BG, time to peak rise for BG, and subjective appetite ratings. In conclusion, all three mixed meals attenuated postprandial glycemic response in comparison to glucose, which may offer advantages to glycemic control.

Εvaluation of oxygen availability on growth and inter-strain interactions of L. monocytogenes in/on liquid, semi-solid and solid laboratory media.

The coexistence and interactions among Listeria monocytogenes strains in combination with the structural characteristics of foods, may influence their growth capacity and thus, the final levels at the time of consumption. In the present study, we aimed to evaluate the effect of oxygen availability in combination with substrate micro-structure on growth and inter-strain interactions of L. monocytogenes. L. monocytogenes strains, selected for resistance to different antibiotics (to enable distinct enumeration), belonging to serotypes 4b (C5, ScottA), 1/2a (6179) and 1/2b (PL25) and were inoculated in liquid (Tryptic Soy Broth supplemented with Yeast Extract - TSB-YE) and solid (TSB-YE supplemented with 0.6% and 1.2% agar) media (2-3 log CFU/mL, g or cm2), single or as two-strain cultures (1:1 strain-ratio). Aerobic conditions (A) were achieved with constant shaking or surface inoculation for liquid and solid media respectively, while static incubation or pour plated media corresponded to hypoxic environment (H). Anoxic conditions (An) were attained by adding 0.1% w/v sodium thioglycolate and paraffin overlay (for solid media). Growth was assessed during storage at 7 °C (n = 3 × 2). Inter-strain interactions were manifested by the difference in the final population between singly and co-cultured strains. Τhe extent of suppression increased with reduction in agar concentration, while the impact of oxygen availability was dependent on strain combination. During co-culture, in liquid and solid media, 6179 was suppressed by C5 by 4.0 (in TSB-YE under H) to 1.8 log units (in solid medium under An), compared to the single culture, which attained population of ca. 9.4 log CFU/mL or g. The growth of 6179 was also inhibited by ScottA by 2.7 and 1.9 log units, in liquid culture under H and An, respectively. Interestingly, in liquid medium under A, H and An, ScottA was suppressed by C5, by 3.3, 2.4 and 2.3 log units, respectively, while in solid media, growth inhibition was less pronounced. Investigating growth interactions in different environments could assist in explaining the dominance of L. monocytogenes certain serotypes.

Assessing the capacity of growth, survival, and acid adaptive response of Listeria monocytogenes during storage of various cheeses and subsequent simulated gastric digestion.

Different physicochemical and microbiological characteristics of cheeses may affect Listeria monocytogenes potential to grow, survive, or exhibit an acid adaptive response during storage and digestion. The objectives of the present study were to assess: i) the survival or growth potential of L.monocytogenes on various cheeses during storage, ii) the effect of initial indigenous microbiota on pathogen growth in comparison to expected growth curves retrieved by existing predictive models, and iii) the impact of habituation on/in cheeses surfaces on the subsequent acid resistance during simulated gastric digestion. Portions of cream (Cottage and Mascarpone), soft (Anthotyros, Camembert, Mastelo®, Manouri, Mozzarella, Ricotta), and semi-hard (Edam, Halloumi, Gouda) cheeses were inoculated with ca. 100CFU/g or cm2 of L.monocytogenes and stored under vacuum or aerobic conditions at 7°C (n=4). The impact of varying (initial) levels of starter culture or indigenous spoilage microbiota on pathogen growth was evaluated by purchasing cheese packages on different dates in relation to production and expiration date (subsequently reflecting to different batches) mimicking a potential situation of cheese contamination with L.monocytogenes during retail display. Values of pH and aw were also monitored and used to simulate growth of L. monocytogenes by existing models and compare it with the observed data of the study. Survival in simulated gastric fluid (SGF) (pH1.5; HCl; max. 120min) was assessed at three time points during storage. Mascarpone, Ricotta, Mozzarella, Camembert, and Halloumi supported L.monocytogenes growth by 0.5-0.8logCFU/g or cm2per day, since low initial levels of total viable counts (TVC) (1.8-3.8logCFU/g or cm2) and high pH/aw values (ca. 6.23-6.64/0.965-0.993) were recorded. On Cottage, Anthotyros, Manouri, Mastelo®, Edam, and Gouda, the pathogen survived at populations similar or lower than the inoculation level due to the high reported competition and/or low pH/aw during storage. L. monocytogenes growth was significantly suppressed (p<0.05) on samples purchased close to expiration date (bearing high TVC), compared to those close to production date, regardless of cheese. Cheeses which supported growth of L.monocytogenes enabled higher survival in gastric acidity along their shelf-life compared to cheeses which did not support growth. However, even in the latter cheeses (i.e., Cottage, Mastelo®, Gouda), total elimination of a persisting low initial contamination was not always achieved. Such findings may provide useful evidence for assessing the risk posed by various cheeses types in relation to their compliance with food safety regulations.

The Economic Crisis in Greece and Its Impact on the Seasonality of Suicides in the Athens Greater Area.

OBJECTIVE: The economic crisis and the implementation of austerity measures in Greece lead to significant socioeconomic changes. The effects of the crisis were mainly felt by the Greek population during the years 2011 and 2012. This study aimed to investigate the impact of Greece's economic crisis on the seasonality of suicides in the Athens Greater Area. METHODS: Data were collected for all recorded cases of suicides committed over a 5-year period (from 2008 to 2012) from the Athens Department of Forensic Medicine. Two sub-periods were studied in relation to the economic crisis: 2008-2010 and 2011-2012. Seasonality was estimated with the Poison regression variant of the circular normal distribution. RESULTS: Suicide seasonality appeared significant during 2008-2010 (relative risk, RR=1.36) and strengthened in the years 2011-2012 (RR=1.69), when the impact of the austerity measures was increasingly being felt by the Greek society. Regarding the latter sub-period, seasonality was established for males (RR=1.75), individuals aged 45 years or more (RR=1.75) and suicide by hanging (RR=1.96). CONCLUSION: The economic crisis in Greece, especially in the period during its effects had a significant impact on the population's economic condition, seems to have strengthened the seasonality of suicides, while a noteworthy suicide risk of 96% was revealed for suicides by hanging (peak in early June).