Interference of gastrointestinal barriers with antibiotic susceptibility of foodborne pathogens: an in vitro case study of ciprofloxacin and tetracycline against Salmonella enterica and Listeria monocytogenes.

Minimum inhibitory concentrations (MIC) assays are often questioned for their representativeness. Especially when foodborne pathogens are tested, it is of crucial importance to also consider parameters of the human digestive system. Hence, the current study aimed to assess the inhibitory capacity of two antibiotics, ciprofloxacin and tetracycline, against Salmonella enterica and Listeria monocytogenes, under representative environmental conditions. More specifically, aspects of the harsh environment of the human gastrointestinal tract (GIT) were gradually added to the experimental conditions starting from simple aerobic lab conditions into an in vitro simulation of the GIT. In this way, the effects of parameters including the anoxic environment, physicochemical conditions of the GIT (low gastric pH, digestive enzymes, bile acids) and the gut microbiota were evaluated. The latter was simulated by including a representative consortium of selected gut bacteria species. In this study, the MIC of the two antibiotics against the relevant foodborne pathogens were established, under the previously mentioned environmental conditions. The results of S. enterica highlighted the importance of the anaerobic environment when conducting such studies, since the pathogen thrived under such conditions. Inclusion of physicochemical barriers led to exactly opposite results for S. enterica and L. monocytogenes since the former became more susceptible to ciprofloxacin while the latter showed lower susceptibility towards tetracycline. Finally, the inclusion of gut bacteria had a bactericidal effect against L. monocytogenes even in the absence of antibiotics, while gut bacteria protected S. enterica from the effect of ciprofloxacin.

Preparation and evaluation of functional cocoa-free spread alternatives from different sources.

Spread products have an important market share as they have high nutritional value and they are increasingly consumed, especially by children as a source of energy. The purpose of this work was to evaluate the potential use of powdered chickpea, black rice, carob, doum, date seeds, and beetroot to produce novel functional spreadable products as cocoa-free alternatives. Additionally, to avoid the side effects of cocoa-based products and to assess the cocoa replacement effects on the sensory properties, chemical composition, texture analysis, viscosity, antioxidant, peroxide stability, and microbial quality during storage periods were compared to the ones of cocoa spread. Sensory evaluation revealed that most formulated spreads were accepted as chocolate spread alternatives since there was no significant difference in overall acceptability among cocoa, chickpea, black rice, carob, and doum, while date seeds and beetroot spreads were significantly less acceptable. A variation was observed in the proximate chemical analysis of the produced functional spreads, as the alternative spreads had different characteristics to each other in their physicochemical, texture, and rheological properties. Results indicated a wide variation in the total phenolic content (TPC) of the different spread extracts. The highest amount of TPC was obtained for beetroot spread (455.84 mg GAE/100 g) followed by black rice spread (436.08 mg GAE/100 g). The obtained results indicated that the antioxidant activity of different spreads was significantly different (p < .05) while based on their microbiological analysis, they could have a shelf life of up to 9 months. According to the results, chickpea, carob, doum, black rice, date seeds, and beetroot powders could be used for the production of cocoa-free alternatives as they were highly acceptable and they showed antioxidant and antimicrobial activity.

Enhancing functional buffalo yogurt: Improving physicochemical properties, biological activities, and shelf life using Marjoram and Geranium essential oils.

The use of essential oils (EOs) has attracted interest in the food industry due to their wide range of beneficial properties. In this study, a new functional yogurt was developed using 2 essential oils [Marjoram (M) and Geranium (G)], at 3 different concentrations (0.2%, 0.4%, and 0.6% vol/vol). The physicochemical properties (syneresis, viscosity, pH, and chemical composition), bioactivities (antioxidant activity, anticancer and antibacterial effects, total phenolic content (TPC), and total flavonoid content (TFC)), and sensory characteristics of the developed yogurt were evaluated. The findings indicated that the yogurts fortified with 0.6% M or G exhibited higher viscosity and lower syneresis compared with other treatments. The yogurt supplemented with 0.6% M displayed significant antibacterial activity against Listeria monocytogenes, Staphylococcus aureus, Salmonella typhimurium, and Escherichia coli. In addition, the yogurt enriched with Geranium and Marjoram oils at a concentration of 0.6% had notably significant (P < 0.05) higher TFC levels compared with the control sample and other concentrations. In the same context, in terms of TPC, yogurt supplemented with 0.6% Marjoram oil displayed significantly (P < 0.05) elevated levels in comparison to the other samples tested. Yogurt enriched with Marjoram oil exhibited noteworthy antioxidant activity, followed by Geranium oil compared with the control samples. The yogurt supplemented with 0.6% M demonstrated strong radical scavenging activity, while the yogurt fortified with 0.6% G showed higher anticancer activity against HepG2 human liver carcinoma cells and oxidative stress enzyme activities. Among the various concentrations of EOs tested, the yogurts fortified with 0.6% M or G EOs exhibited the most favorable outcomes, followed by 0.4% M or G. To summarize, G and M EOs can be used as a potential nutritious ingredient and as a natural preservative for milk and related products.

Novel methods to monitor the biodegradation of polylactic acid (PLA) by Amycolatopsis orientalis and Amycolatopsis thailandensis.

Plastics are essential in modern life, but their conventional production is problematic due to environmental pollution and waste management issues. Polylactic acid (PLA) is a widely used bioplastic that is bio-based and biodegradable, making it a key player in the bioeconomy. PLA has been proven to be degradable in various settings, including aqueous, soil, and compost environments. However, monitoring and optimizing PLA biodegradation remains challenging. This study proposes methods to improve the quantification of PLA biodegradation by Amycolatopsis spp. Ultrasound treatments (10 s) significantly improved the enumeration of viable Amycolatopsis cells by breaking the pellets into quantifiable individual cells. A separation technique combining ultrasound (120 s) and 40 µm cell strainers effectively isolated PLA particles from biomass to quantify PLA weight loss. This enabled the monitoring of PLA biofragmentation. Finally, CO2 production was measured according to ISO 14852 to quantify mineralization. Integrating these methods provides an improved quantification for PLA biodegradation along its different stages. In a case study, this led to the construction of a carbon balance where 85.1% of initial carbon content was successfully tracked. The developed techniques for monitoring of PLA biodegradation are essential to design future waste management strategies for biodegradable plastics.

Radio frequency inactivation of Salmonella Typhimurium and Listeria monocytogenes in skimmed and whole milk powder.

Milk powder is a convenient, shelf-stable food ingredient used in a variety of food products. However, pathogenic bacteria can be present and survive during prolonged storage, leading to outbreaks of foodborne diseases and product recalls. Radio frequency (RF) heating is a processing technology suitable for bulk treatment of milk powder, aiming at microbial inactivation. This study investigates the RF inactivation of Salmonella Typhimurium and Listeria monocytogenes in two types of milk powder; skimmed and whole milk powder. Specifically, the aims were to (i) examine the influence of the powder's composition on bacterial inactivation, (ii) evaluate the response of bacteria with different Gram properties (Gram positive and Gram negative) and (iii) verify the use of Enterococcus faecium as a surrogate for the two microorganisms for the specific RF process. In order to examine exclusively the influence of RF, a non-isothermal temperature profile was used, employing solely different RF energy levels to heat the product to the target temperatures. A log-linear model with a Bigelow-type temperature dependency was fitted to the experimental data. S. Typhimurium was less susceptible to RF treatments in comparison to L.monocytogenes, demonstrating a higher inactivation rate (k) and higher percentage of sublethal injury. A higher k was also observed for both microorganisms in the whole milk powder, indicating that the increased fat content and decreased levels of lactose and protein in the milk powder had an adverse impact on the microbial survival for both pathogens. The surrogate microorganism E. faecium successfully validated the microbial response of the two microorganisms to RF treatments. In general, a low heating rate RF-only process was successful in inactivating the two foodborne pathogens in skimmed and whole milk powder by 4 log(CFU/g).

Gut microbiota of the small intestine as an antimicrobial barrier against foodborne pathogens: Impact of diet on the survival of S. Typhimurium and L. monocytogenes during in vitro digestion.

The human gastrointestinal tract employs an assortment of chemical, enzymatic and immune barriers to impede pathogen colonization. An essential component of these barriers is the gut microbiota, which infers protection against ingested pathogens through its colonization resistance mechanisms. Specifically, the gut microbiota of the distal small intestine (ileum) renders a crucial line of defense, given that this location is regarded as an important interaction site. This study aimed to evaluate the impact of the ileal microbiota on the survival of the foodborne pathogens Salmonella enterica serotype Typhimurium and Listeria monocytogenes, utilizing an in vitro digestion model system. Moreover, the effect of diet on the gut microbiota colonization resistance mechanisms was assessed, by comparing a healthy (high fiber/low sugar) and a western diet (low fiber/high sugar). For S. Typhimurium, the results revealed that the digestion of a healthy diet led to a similar inactivation compared to the western diet, with the values of total log reduction being 0.83 and 0.82 log(CFU), respectively; yet the lack of readily accessible nutrients in the healthy diet combined with the acidic shock during gastric digestion caused the induction of stress tolerance to the pathogen. This resulted in increased pathogen survival in the presence of gut microbiota, with S. Typhimurium proliferating during the ileal phase with a maximum specific growth rate of 0.16 1/h. On the contrary, for L. monocytogenes, the healthy diet was associated with a greater inactivation than the western diet (total log reduction values: 3.08 and 1.30 log(CFU), respectively), which appeared strongly influenced by the encounter of the pathogen with the gut microbiota. Regarding the latter, the species Escherichia coli and Bacteroides thetaiotaomicron appeared to be the most prevalent in most cases. Finally, it was also demonstrated that the ileal microbiota colonization resistance mechanisms largely relied on competitive responses. The obtained knowledge of this research can contribute to the development and/or complementation of defensive strategies against pathogen infection, while also underlining the value of in vitro approaches.

Macroscopic modeling of the growth and substrate consumption of wild type and genetically modified Pichia pastoris.

Pichia pastoris is a popular yeast platform to generate several industrially relevant products which have applications in a wide range of sectors. The complexities in the processes due to the addition of a foreign gene are not widely explored. Since these complexities can be dependent on the strain characteristics, promoter, and type of protein produced, it is vital to investigate the growth and substrate consumption patterns of the host to facilitate customized process optimization. In this study, the growth rates of P. pastoris GS115 wild type (WT) and genetically modified (GM) strains grown on glycerol and methanol in batch cultivation mode were estimated and the model providing the best representation of the true growth kinetics based on substrate consumption was identified. It was observed that the growth of P. pastoris exhibits Haldane kinetics on glycerol rather than the most commonly used Monod kinetics due to the inability of the latter to describe growth inhibition at high concentrations of glycerol. Whereas, the cardinal parameter model, a newly proposed model for this application, was found to be the best fitting to describe the growth of P. pastoris on methanol due to its ability to describe methanol toxicity. Interestingly, the findings from this study concluded that in both substrates, the genetically engineered strain exhibited a higher growth rate compared to the WT strain. Such an observation has not been established yet in other published works, indicating an opportunity to further optimize the carbon source feeding strategies when the host is grown in fed-batch mode.

Influence of microbial cell morphology and composition on radio frequency heating of simple media at different frequencies.

The effect of Listeria monocytogenes, Salmonella Typhimurium, and Saccharomyces cerevisiae on RF heating was studied in sterilized Milli-Q water and saline solution during treatments at 27.0 ± 0.6 MHz and 3.0 ± 0.02 MHz for 30 min. The presence of microorganisms caused a significant increase in temperature (maximum to 54.9 °C), with no significant decrease in cell numbers being observed for any conditions. For both media and frequencies, heating rates followed the order S. Typhimurium ≤ L. monocytogenes ≤ S. cerevisiae, except for heating at 3.0 ± 0.02 MHz in saline solution, where heating rates for S. cerevisiae and S. Typhimurium were equal. Generally, heating rates for microorganisms were significantly higher at 27.0 ± 0.6 MHz than at 3.0 ± 0.02 MHz, except for the S. cerevisiae case. Observed phenomena were probably caused by differences in the cell lipid and peptidoglycan content, with interaction effects with salt being present. This study was the first to investigate the influence of the presence of microorganisms on heating behavior of simple media. On the long term, more research on this topic could lead to finding specific RF frequencies more suitable for the heating of specific media and products for various applications.

Probiotic Properties and Antioxidant Activity In Vitro of Lactic Acid Bacteria.

The properties of probiotics such as lactic acid bacteria (LAB) have been widely studied over the last decades. In the present study, four different LAB species, namely Lactobacillus gasseri ATCC 33323, Lacticaseibacillus rhamnosus GG ATCC 53103, Levilactobacillus brevis ATCC 8287, and Lactiplantibacillus plantarum ATCC 14917, were investigated in order to determine their ability to survive in the human gut. They were evaluated based on their tolerance to acids, resistance to simulated gastrointestinal conditions, antibiotic resistance, and the identification of genes encoding bacteriocin production. All four tested strains demonstrated high resistance to simulated gastric juice after 3 h, and the viable counts revealed declines in cell concentrations of less than 1 log cycle. L. plantarum showed the highest level of survival in the human gut, with counts of 7.09 log CFU/mL. For the species L. rhamnosus and L. brevis, the values were 6.97 and 6.52, respectively. L. gasseri, after 12 h, showed a 3.96 log cycle drop in viable counts. None of the evaluated strains inhibited resistance to ampicillin, gentamicin, kanamycin, streptomycin, erythromycin, clindamycin, tetracycline, or chloramphenicol. With regard to bacteriocin genes, the Pediocin PA gene was identified in Lactiplantibacillus plantarum ATCC 14917, Lacticaseibacillus rhamnosus GG ATCC 53103, and Lactobacillus gasseri ATCC 33323. The PlnEF gene was detected in Lactiplantibacillus plantarum ATCC 14917 and Lacticaseibacillus rhamnosus GG ATCC 53103. The Brevicin 174A and PlnA genes were not detected in any bacteria. Moreover, the potential antioxidant activity of LAB's metabolites was evaluated. At the same time, the possible antioxidant activity of metabolites of LAB was first tested using the free radical DDPH• (a, a-Diphenyl-ß-Picrylhydrazyl) and then evaluated with regard to their radical scavenging activity and inhibition against peroxyl radical induced DNA scission. All strains showed antioxidant activity; however, the best antioxidant activity was achieved by L. brevis (94.47%) and L. gasseri (91.29%) at 210 min. This study provides a comprehensive approach to the action of these LAB and their use in the food industry.

Influence of Hurdle Technology on Foodborne Pathogen Survival in the Human Gastrointestinal Tract.

The application of several sublethal stresses in hurdle technology can exert microbial stress resistance, which, in turn, might enable foodborne pathogens to overcome other types of lethal stresses, such as the gastrointestinal barriers. The present study evaluated the survival of Salmonella Typhimurium and Listeria monocytogenes during simulated digestion, following exposure to combinations of water activity (aw), pH and storage temperature stresses. The results revealed that both pathogens survived their passage through the simulated gastrointestinal tract (GIT) with their previous habituation to certain hurdle combinations inducing stress tolerance. More specifically, the habituation to a low temperature or to a high pH resulted in the increased stress tolerance of Salmonella, while for Listeria, the cells appeared stress tolerant after exposure to a high temperature or to a low pH. Nonetheless, both pathogens expressed increased sensitivity after habituation to growth-limiting hurdle combinations. The survival of stress-tolerant pathogenic cells in the human GIT poses major public health issues, since it can lead to host infection. Consequently, further research is required to obtain a deeper understanding of the adaptive stress responses of foodborne bacteria after exposure to combinations of sublethal hurdles to improve the existing food safety systems.

Effect of food structure and buffering capacity on pathogen survival during in vitro digestion.

Even though a plethora of barriers are employed by the human gastrointestinal tract (GIT) to cope with invading pathogens, foodborne diseases are still a common problem. The survival of food pathogens in the GIT is known to depend on food carrier properties. The aim of this study was to investigate the influence of food buffering capacity and food structure on the survival of Salmonella Typhimurium and Listeria monocytogenes during simulated digestion, following contamination of different food model systems that had different combinations of fat and protein content. The results illustrated the strong protective properties of proteins, acting either as a strong buffering agent or as a physical barrier against gastric acidity, for both pathogens. In comparison, fat manifested a lower buffering capacity and weaker protective effects against the two pathogens. Intriguingly, a low fat content was often linked with increased microbial resistance. Nonetheless, both pathogens survived their transit through the simulated GIT in all cases, with S. Typhimurium exhibiting growth during intestinal digestion and L.monocytogenes demonstrating a healthy residual population at the end of the intestinal phase. These results corroborate the need for a deeper understanding regarding the mechanisms with which food affects bacterial survival in the human GIT.

The role of temperature and carbon dioxide climatic stress factors on the growth kinetics of Escherichia coli.

AIMS: The global level of carbon dioxide and temperature in the atmosphere is expected to increase, which may affect the survival of the stress-adapted bacteria. In this study, the effect of temperature and dissolved carbon dioxide on the growth rate of Escherichia coli-eGFP tagged strain was studied, thus assessing its response to induced environmental stress factors. METHODS AND RESULTS: A kinetic assay has been performed using a microplate reader with a spectrofluorometer to determine the specific growth rates. Polynomial models were developed to correlate the environmental conditions of temperature and carbon dioxide with Escherichia coli BL21 (DE3) growth in culture media and dairy by-products. At a temperature of 42°C, as the dissolved CO2 increased, a decrease in µmax by 0.76 h-1 was observed. In contrast, at 27°C, this increase led to an increase in µmax by 0.99 h-1. Moreover, a correction factor was added when applying the model to dairy whey samples. CONCLUSIONS: The application of this developed model can be considered a useful tool for predicting the growth of Escherichia coli using climate projections.

The territorial nature of aggression in biofilms.

Microbial conflicts have a particularly aggressive nature. In addition to other chemical, mechanical, and biological weapons in their repertoire, bacteria have evolved bacteriocins, which are narrow-spectrum toxins that kill closely related strains. Bacterial cells are known to frequently use their arsenal while competing against each other for nutrients and space. This stands in contrast with the animal world, where conflicts over resources and mating opportunities are far less lethal, and get commonly resolved via ritualized fighting or "limited war" tactics. Prevalence of aggression in microbial communities is usually explained as due to their limited ability to resolve conflicts via signaling as well as their limited ability to pull out from conflicts due to the sessile nature of their life within biofilms. We use an approach that combines Evolutionary Game Theory (EGT) and Individual-based Modeling (IbM) to investigate the origins of aggression in microbial conflicts. In order to understand how the spatial mode of growth affects the cost of a fight, we compare the growth dynamics emerging from engaging in aggression in a well-mixed system to a spatially structured system. To this end, a mathematical model is constructed for the competition between two bacterial strains where each strain produces a diffusible toxin to which the other strain is sensitive. It is observed that in the biofilm growth mode, starting from a mixed layer of two strains, mutual aggression gives rise to an exceedingly high level of spatial segregation, which in turn reduces the cost of aggression on both strains compared to when the same competition occurs in a well-mixed culture. Another observation is that the transition from a mixed layer to segregated growth is characterized by a switch in the overall growth dynamics. An increased "lag time" is observed in the overall population growth curve that is associated with the earlier stages of growth, when each strain is still experiencing the inhibiting effect of the toxin produced by its competitor. Afterwards, an exponential phase of growth kicks in once the competing strains start segregating from each other. The emerging "lag time" arises from the spiteful interactions between the two strains rather than acclimation of cells' internal physiology. Our analysis highlights the territorial nature of microbial conflicts as the key driver to their elevated levels of aggression as it increases the benefit-to-cost ratio of participating in antagonistic interactions.

Processing Method for the Quantification of Methanol and Ethanol from Bioreactor Samples Using Gas Chromatography-Flame Ionization Detection.

Methanol, a simple polar solvent, has been widely identified as an attractive carbon source to produce chemicals and fuels in bioprocesses. Specifically, to achieve recombinant protein production from methylotrophic yeasts, such as Pichia pastoris, this organic solvent can be used as a sole carbon source for growth and maintenance as well as an inducer for protein expression. However, if methanol feeding is not controlled well in such a fermentation process, accumulation of the solvent in the growth media will have a detrimental effect on the cells. Hence, monitoring the levels of methanol in these fermentation processes is a crucial step to ensure a healthy culture and maximum protein production. There are various techniques elaborated in the literature for monitoring methanol in cell cultures, but often, they appear to be expensive methods that are less affordable for many laboratories. This is because, in addition to the sophisticated equipment that is required for the analysis, the complexity of the samples retrieved from the bioprocesses necessitates laborious processing steps often involving expensive tools. In this study, a fast, simple, and sensitive method is developed to process biological samples by using the salting-out-assisted liquid-liquid extraction technique to quantify the concentration of methanol and ethanol using gas chromatography. On comparing the combinations of widely available salts and solvents, it was noticed that salting out using potassium carbonate followed by the liquid-liquid extraction of the analyte using ethyl acetate showed the best recovery. Followed by this, a validation test for the developed method was performed, which resulted in good peak resolution, linearity, and limit of detection for the quantitation of methanol and ethanol. By further assessing the tested combination, it was confirmed that its application could be extended to other matrices. Such an approach facilitates the possibility to monitor and control the methanol levels in fermentation and aids in bioprocess optimization.

Enhancing the biodegradation of (bio)plastic through pretreatments: A critical review.

As plastic packaging becomes nearly indispensable in the plastic economy, rigorous efforts have been made to recapture the material value form this waste stream, which is mostly composed of highly resistant plastics. Biodegradation offers an attractive alternative for conventional plastic waste treatment as this approach is environmentally friendly, has low cost and facilitates valorisation. Moreover, there is also an increasing interest in plastic pretreatments waste to enhance biodegradation. This review investigates the pretreatment methods that optimise plastic biodegradation by examining the process's mechanisms and key influencing factors, which can be categorised into: biotic factors, abiotic factors and polymer characteristics. Various types of chemical and physical pretreatments have demonstrated to effectively enhance biodegradation through oxidation and surface changes on the plastics, leading to increased bioconversion rates and biogas production. A critical evaluation of the various categories of pretreatment methods is presented. This evaluation leads to the conclusion that the category of non-thermal physical treatments is most promising, due to the relatively low energy requirements and the absence of a need for chemical additions. Moreover, non-thermal physical treatments have demonstrated application potential at large scale. Based on these conclusions, pretreatments are expected to be an integral part of the biodegradation of plastics within a circular economy approach.

A protocol for the cultivation and monitoring of ileal gut microbiota surrogates.

AIMS: This research aimed to develop and validate a cultivation and monitoring protocol that is suitable for a surrogate microbial community that accounts for the gut microbiota of the ileum of the small intestine. METHODS AND RESULTS: Five bacterial species have been selected as representatives of the ileal gut microbiota and a general anaerobic medium (MS-BHI, as minimally supplemented brain heart infusion) has been constructed and validated against BCCM/LGM recommended and commercial media. Moreover, appropriate selective/differential media have been investigated for monitoring each ileal gut microbiota surrogate. Results showed that MS-BHI was highly efficient in displaying individual and collective behaviour of the ileal gut microbiota species, when compared with other types of media. Likewise, the selective/differential media managed to identify and describe the behaviour of their targeted species. CONCLUSIONS: MS-BHI renders a highly efficient, inexpensive and easy-to-prepare cultivation and enumeration alternative for the surrogate ileal microbiota species. Additionally, the selective/differential media can identify and quantify the bacteria of the surrogate ileal microbial community. SIGNIFICANCE AND IMPACT OF STUDY: The selected gut microbiota species can represent an in vitro ileal community, forming the basis for future studies on small intestinal microbiota. MS-BHI and the proposed monitoring protocol can be used as a standard for gut microbiota studies that utilize conventional microbiological techniques.

Effects of Temperature and pH on Recombinant Thaumatin II Production by Pichia pastoris.

The sweet protein thaumatin is emerging as a promising sugar replacer in the market today, especially in the food and beverage sector. Rising demand for its production necessitates the large-scale extraction of this protein from its natural plant source, which can be limited in terms of raw material availability and production costs. Using a recombinant production technique via a yeast platform, specifically, Pichia pastoris, is more promising to achieve the product economically while maintaining batch-to-batch consistency. However, the bioproduction of recombinant proteins requires the identification of optimal process variables, constituting the maximal yield of the product of interest. These variables have a direct effect on the growth of the host organism and the secretion levels of the recombinant protein. In this study, two important environmental factors, pH, and temperature were assessed by cultivating P. pastoris in shake flasks to understand their influence on growth and the production levels of thaumatin II protein. The results from the pH study indicate that P. pastoris attained a higher viable cell density and secretion of protein at pH 6.0 compared to 5.0 when grown at 30 °C. Furthermore, within the three levels of temperatures investigated when grown at pH 6.0, the protein levels were the highest at 30 °C compared to 20 and 25 °C, whereas 25 °C exhibited the highest viable cell density. Interestingly, the trend observed from the qualitative effects of temperature and pH occurred in all the media that was investigated. These results broaden our understanding of how pH and temperature adjustment during P. pastoris cultivation aid in enhancing the production yields of thaumatin II prior to optimising the fed batch bioreactor operation.

Behavior of the Surviving Population of Listeria monocytogenes and Salmonella Typhimurium Biofilms Following a Direct Helium-Based Cold Atmospheric Plasma Treatment.

Although the Cold Atmospheric Plasma (CAP) technology proved promising for inactivation of biofilms present on abiotic food contact surfaces, more research is required to examine the behavior of the CAP surviving biofilm-associated cells. It was therefore examined whether (i) CAP treated (Listeria monocytogenes and Salmonella Typhimurium) biofilm-associated cells were able to further colonize the already established biofilms during a subsequent incubation period and (ii) isolates of the surviving population became less susceptible toward CAP when the number of biofilm development-CAP treatment cycles increased. For this purpose, a direct treatment was applied using a helium-based Dielectric Barrier Discharge electrode configuration. Results indicated that the surviving population was able to further colonize the already established biofilms, since the cell density of the CAP treated + incubated biofilms equaled the initial density of the untreated biofilms. For the L. monocytogenes biofilms, also the total biomass proved to further increase, which might result in an even further increased resistance. The susceptibility of the biofilm-associated cells proved to be influenced by the specific number of CAP treatment cycles, which might potentially result in an overestimation of the CAP treatment efficacy and, consequently, an increased risk of food contamination.

Dishonest Signaling in Microbial Conflicts.

Quorum sensing is a cell-cell communication system that bacteria use to express social phenotypes, such as the production of extracellular enzymes or toxins, at high cell densities when these phenotypes are most beneficial. However, many bacterial strains are known to lack a sensing mechanism for quorum signals, despite having the gene responsible for releasing the signals to the environment. The aim of this article is 2-fold. First, we utilize mathematical modeling and signaling theory to elucidate the advantage that a bacterial species can gain by releasing quorum signals, while not being able to sense them, in the context of ecological competition with a focal quorum sensing species, by reducing the focal species' ability to optimize the timing of expression of the quorum sensing regulated phenotype. Additionally, the consequences of such "dishonest signaling," signaling that has evolved to harm the signal's receiver, on the focal quorum sensing species are investigated. It is found that quorum sensing bacteria would have to incur an additional, strategic, signaling cost in order to not suffer a reduction in fitness against dishonest signaling strains. Also, the concept of the Least Expensive Reliable Signal is introduced and applied to study how the properties of the regulated phenotype affect the metabolic investment in signaling needed by the quorum sensing bacteria to withstand dishonest signaling.

Characterization of Bacterial Microbiota of P.D.O. Feta Cheese by 16S Metagenomic Analysis.

BACKGROUND: The identification of bacterial species in fermented PDO (protected designation of origin) cheese is important since they contribute significantly to the final organoleptic properties, the ripening process, the shelf life, the safety and the overall quality of cheese. METHODS: Ten commercial PDO feta cheeses from two geographic regions of Greece, Epirus and Thessaly, were analyzed by 16S metagenomic analysis. RESULTS: The biodiversity of all the tested feta cheese samples consisted of five phyla, 17 families, 38 genera and 59 bacterial species. The dominant phylum identified was Firmicutes (49% of the species), followed by Proteobacteria (39% of the species), Bacteroidetes (7% of the species), Actinobacteria (4% of the species) and Tenericutes (1% of the species). Streptococcaceae and Lactobacillaceae were the most abundant families, in which starter cultures of lactic acid bacteria (LAB) belonged, but also 21 nonstarter lactic acid bacteria (NSLAB) were identified. Both geographical areas showed a distinctive microbiota fingerprint, which was ultimately overlapped by the application of starter cultures. In the rare biosphere of the feta cheese, Zobellella taiwanensis and Vibrio diazotrophicus, two Gram-negative bacteria which were not previously reported in dairy samples, were identified. CONCLUSIONS: The application of high-throughput DNA sequencing may provide a detailed microbial profile of commercial feta cheese produced with pasteurized milk.