Isolation, identification and screening of yeasts towards their ability to assimilate biodiesel-derived crude glycerol: microbial production of polyols, endopolysaccharides and lipid.

AIMS: To assess the ability of various newly isolated or belonging in official collections yeast strains to convert biodiesel-derived glycerol (Gly) into added-value compounds. METHODS AND RESULTS: Ten newly isolated yeast strains belonging to Debaryomyces sp., Naganishia uzbekistanensis, Rhodotorula sp. and Yarrowia lipolytica, isolated from fishes, metabolized Gly under nitrogen limitation. The aim of the study was to identify potential newly isolated microbial candidates that could produce single-cell oil (SCO), endopolysaccharides and polyols when these micro-organisms were grown on biodiesel-derived Gly. As controls producing SCO and endopolysaccharides were the strains Rhodotorula glutinis NRRL YB-252 and Cryptococcus curvatus NRRL Y-1511. At initial Gly (Gly0 ) ≈40 g l-1 , most strains presented remarkable dry cell weight (DCW) production, whereas Y. lipolytica and Debaryomyces sp. produced non-negligible quantities of mannitol and arabitol (Ara). Five strains were further cultivated at increasing Gly0 concentrations. Rhodotorula glutinis NRRL YB-252 produced 7·2 g l-1 of lipid (lipid in DCW value ≈38% w/w), whereas Debaryomyces sp. FMCC Y69 in batch-bioreactor experiment with Gly0 ≈80 g l-1 , produced 30-33 g l-1 of DCW and ~30 g l-1 of Ara. At shake-flasks with Gly0 ≈125 g l-1 , Ara of ~48 g l-1 (conversion yield of polyol on Gly consumed ≈0·62 g g-1 ) was achieved. Cellular lipids of all yeasts contained in variable concentrations oleic, palmitic, stearic and linoleic acids. CONCLUSIONS: Newly isolated, food-derived and non-previously studied yeast isolates converted biodiesel-derived Gly into several added-value metabolites. SIGNIFICANCE AND IMPACT OF THE STUDY: Alternative ways of crude Gly valorization through yeast fermentations were provided and added-value compounds were synthesized.

Effect of inoculum size, bacterial species, type of surfaces and contact time to the transfer of foodborne pathogens from inoculated to non-inoculated beef fillets via food processing surfaces.

The objective of the present study was to determine the factors affecting the transfer of foodborne pathogens from inoculated beef fillets to non-inoculated ones, through food processing surfaces. Three different levels of inoculation of beef fillets surface were prepared: a high one of approximately 107 CFU/cm2, a medium one of 105 CFU/cm2 and a low one of 103 CFU/cm2, using mixed-strains of Listeria monocytogenes, or Salmonella enterica Typhimurium, or Escherichia coli O157:H7. The inoculated fillets were then placed on 3 different types of surfaces (stainless steel-SS, polyethylene-PE and wood-WD), for 1 or 15 min. Subsequently, these fillets were removed from the cutting boards and six sequential non-inoculated fillets were placed on the same surfaces for the same period of time. All non-inoculated fillets were contaminated with a progressive reduction trend of each pathogen's population level from the inoculated fillets to the sixth non-inoculated ones that got in contact with the surfaces, and regardless the initial inoculum, a reduction of approximately 2 log CFU/g between inoculated and 1st non-inoculated fillet was observed. S. Typhimurium was transferred at lower mean population (2.39 log CFU/g) to contaminated fillets than E. coli O157:H7 (2.93 log CFU/g), followed by L. monocytogenes (3.12 log CFU/g; P < 0.05). Wooden surfaces (2.77 log CFU/g) enhanced the transfer of bacteria to subsequent fillets compared to other materials (2.66 log CFU/g for SS and PE; P < 0.05). Cross-contamination between meat and surfaces is a multifactorial process strongly depended on the species, initial contamination level, kind of surface, contact time and the number of subsequent fillet, according to analysis of variance. Thus, quantifying the cross-contamination risk associated with various steps of meat processing and food establishments or households can provide a scientific basis for risk management of such products.

Factors affecting quality and safety of fresh-cut produce.

The quality of fresh-cut fruit and vegetable products includes a combination of attributes, such as appearance, texture, and flavor, as well as nutritional and safety aspects that determine their value to the consumer. Nutritionally, fruit and vegetables represent a good source of vitamins, minerals, and dietary fiber, and fresh-cut produce satisfies consumer demand for freshly prepared, convenient, healthy food. However, fresh-cut produce deteriorates faster than corresponding intact produce, as a result of damage caused by minimal processing, which accelerates many physiological changes that lead to a reduction in produce quality and shelf-life. The symptoms of produce deterioration include discoloration, increased oxidative browning at cut surfaces, flaccidity as a result of loss of water, and decreased nutritional value. Damaged plant tissues also represent a better substrate for growth of microorganisms, including spoilage microorganisms and foodborne pathogens. The risk of pathogen contamination and growth is one of the main safety concerns associated with fresh-cut produce, as highlighted by the increasing number of produce-linked foodborne outbreaks in recent years. The pathogens of major concern in fresh-cut produce are Listeria monocytogenes, pathogenic Escherichia coli mainly O157:H7, and Salmonella spp. This article describes the quality of fresh-cut produce, factors affecting quality, and various techniques for evaluating quality. In addition, the microbiological safety of fresh-cut produce and factors affecting pathogen survival and growth on fresh-cut produce are discussed in detail.

A study on the implications of NaCl reduction in the fermentation profile of Conservolea natural black olives.

This study examined the impact of different mixtures of NaCl, KCl, and CaCl(2) on the fermentation profiles of Conservolea natural black olives. Five different combinations of chloride salts were investigated, namely (i) 8% NaCl (control treatment), (ii) 4% NaCl and 4% KCl, (iii) 4% NaCl and 4% CaCl(2), (iv) 4% KCl and 4% CaCl(2), and (v) 2.6% NaCl-2.6% KCl-2.6% CaCl(2). The changes in the microbial association (lactic acid bacteria, yeasts, Enterobacteriaceae), pH, titratable acidity, organic acids, volatile compounds, and mineral content in olive flesh were analyzed. Results demonstrated that all salt combinations led to vigorous lactic acid processes based on the obtained values of pH (3.9-4.2) and titratable acidity (0.70-0.86 g lactic acid per 100 ml brine). Organoleptic evaluation was a critical factor in the acceptability of the final product. Increasing concentrations of CaCl(2) or a combination of KCl and CaCl(2) rendered the product bitter with low acceptability by the taste panel. Only one combination of chloride salts (4% NaCl and 4% KCl) could finally produce olives with lower sodium content and good organoleptic attributes. The results of this study could be employed by the Greek table olive industry in an attempt to produce natural black olives with less sodium without affecting the traditional taste of fermented olives in order to meet consumers' demand for low sodium dietary intake.

Use of titanium dioxide (TiO2) photocatalysts as alternative means for Listeria monocytogenes biofilm disinfection in food processing.

The aim of this work was to study the photocatalytic activity of titanium dioxide (TiO(2)) against Listeria monocytogenes bacterial biofilm. Different TiO(2) nanostructured thin films were deposited on surfaces such as stainless steel and glass using the doctor-blade technique. All the surfaces were placed in test tubes containing Brain Heart (BH) broth and inoculated with L. monocytogenes. Test tubes were then incubated for 10 days at 16°C in order to allow biofilm development. After biofilm formation, the surfaces were illuminated by ultraviolet A light (UVA; wavelength of 315-400 nm). The quantification of biofilms was performed using the bead vortexing method, followed by agar plating and/or by conductance measurements (via the metabolic activity of biofilm cells). The presence of the TiO(2) nanoparticles resulted in a fastest log-reduction of bacterial biofilm compared to the control test. The biofilm of L. monocytogenes for the glass nanoparticle 1 (glass surface modified by 16% w/v TiO(2)) was found to have decreased by 3 log CFU/cm(2) after 90 min irradiation by UVA. The use of TiO(2) nanostructured photocatalysts as alternative means of disinfecting contaminated surfaces presents an intriguing case, which by further development may provide potent disinfecting solutions. Surface modification using nanostructured titania and UV irradiation is an innovative combination to enhance food safety and economizing time and money.

Yeast heterogeneity during spontaneous fermentation of black Conservolea olives in different brine solutions.

AIMS: To assess the yeast community structure and dynamics during Greek-style processing of natural black Conservolea olives in different brine solutions. METHODS AND RESULTS: Black olives were subjected to spontaneous fermentation in 6% (w/v) NaCl brine solution or brine supplemented with (i) 0.5% (w/v) glucose, (ii) 0.2% (v/v) lactic acid and (iii) both glucose and lactic acid. Yeast species diversity was evaluated at the early (2 days), middle (17 days) and final (35 days) stages of fermentation by restriction fragment length polymorphism and sequence analyses of the 5.8S internal transcribed spacer and the D1/D2 ribosomal DNA (rDNA) regions of isolates. Analysis revealed a relatively broad range of biodiversity composed of 10 genera and 17 species. In all treatments, yeasts were the main micro-organisms involved in fermentation together with lactic acid bacteria that coexisted throughout the processes. Metschnikowia pulcherrima was the dominant yeast species at the onset of fermentation, followed by Debaryomyces hansenii and Aureobasidium pullulans. Species heterogeneity changed as fermentations proceeded and Pichia membranifaciens along with Pichia anomala evolved as the main yeasts of olive elaboration, prevailing at 17 and 35 days of the process. Molecular techniques allowed for the identification of five yeast species, namely A. pullulans, Candida sp., Candida silvae, Cystofilobasidium capitatum and M. pulcherrima, which have not been reported previously in black olive fermentation. CONCLUSIONS: By using molecular techniques, a rich yeast community was identified from Conservolea black olive fermentations. Metschnikowia pulcherrima was reported for the first time to dominate in different brines at the onset of fermentation, whereas Pichia anomala and P. membranifaciens evolved during the course. The addition of glucose and/or lactic acid perturbed yeast succession and dominance during fermentation. SIGNIFICANCE AND IMPACT OF THE STUDY: Yeasts have an important role in black olive fermentation and contribute to the development of the organoleptic characteristics of the final product. At the same time, certain species can cause significant spoilage. The present study adds to a better knowledge of yeast communities residing in olive fermentations towards a well-controlled process with minimization of product's losses.

Effect of microbial cell-free meat extract on the growth of spoilage bacteria.

AIMS: This study examined the effect of microbial cell-free meat extract (CFME) derived from spoiled meat, in which quorum sensing (QS) compounds were present, on the growth kinetics (lag phase, and growth rate) of two spoilage bacteria, Pseudomonas fluorescens and Serratia marcescens. METHODS AND RESULTS: Aliquots of CFME from spoiled meat were transferred to Brain Heart Infusion broth inoculated with 10(3) CFU ml(-1) of 18 h cultures of Ps. fluorescens or Ser. marcescens, both fresh meat isolates; CFME derived from unspoiled fresh meat ('clean' meat) served as a control. Changes in impedance measurements were monitored for 48 h, and the detection time (Tdet) was recorded. It was found that in the absence of CFME containing QS compounds the Tdet was shorter (P < 0.05) than that in broth samples with added CFME from spoiled meat. The rate of growth of Ps. fluorescens, recorded as the maximum slope rate of conductance changes (MSrCC), after Tdet, was higher (P < 0.05) in samples with CFME containing QS compounds compared to samples without CFME or CFME derived from 'clean' meat. Similar results in MSrCC of impedance changes were obtained for Ser. marcescens. CONCLUSIONS: The study indicated that the growth rate (expressed in MSrCC units) of meat spoilage bacteria in vitro was enhanced in samples supplemented with CFME containing QS compounds compared to control samples (i.e., without CFME or with CFME from 'clean' meat). This behaviour may explain the dominant role of these two bacteria in the spoilage of meat. SIGNIFICANCE AND IMPACT OF THE STUDY: These results illustrate the potential effect of signalling compounds released during storage of meat on the behaviour of meat spoilage bacteria. Understanding such interactions may assist in the control of fresh meat quality and the extension of its shelf life.

Disinfectant test against monoculture and mixed-culture biofilms composed of technological, spoilage and pathogenic bacteria: bactericidal effect of essential oil and hydrosol of Satureja thymbra and comparison with standard acid-base sanitizers.

AIMS: To assess the antimicrobial action of three natural-derived products (essential oil, decoction and hydrosol of Satureja thymbra) against biofilms, composed of useful, spoilage and pathogenic bacteria (formed as monoculture or/and mixed-culture), and to compare their efficiency with three standard acid and alkaline chemical disinfectants. METHODS AND RESULTS: Two acids (hydrochloric and lactic, pH 3), one alkali (sodium hydroxide, pH 11), the essential oil of S. thymbra (1% v/v) and the two by-products of the essential oil purification procedure (the decoction and the hydrosol fraction of essential oil, 100%), were tested against biofilms formed by five bacterial species, either as monospecies, or as mixed-culture of all species. The tested bacterial species were Staphylococcus simulans and Lactobacillus fermentum (useful technological bacteria), Pseudomonas putida (spoilage bacterium), Salmonella enterica and Listeria monocytogenes (pathogenic bacteria). Biofilms were left to be formed on stainless steel coupons for 5 days at 16 degrees C, before the application of disinfection treatments, for 60 and 180 min. The disinfection efficiency was evaluated by detaching the remaining viable biofilm cells and enumerating them by agar plating, as well as by automated conductance measurements (using Rapid Automated Bacterial Impedance Technique). Both these methods revealed that the essential oil and the hydrosol of S. thymbra exhibited a strong antimicrobial action against both monospecies and mixed-culture biofilms. Surprisingly, the efficiency of the other three acid-base disinfectants was not adequate, although a long antimicrobial treatment was applied (180 min). CONCLUSIONS: The essential oil of S. thymbra (1%), as well as its hydrosol fraction (100%), presents sufficient bactericidal effect on bacterial biofilms formed on stainless steel. SIGNIFICANCE AND IMPACT OF THE STUDY: Use of natural antimicrobial agents could provide alternative or supplemented ways for the disinfection of microbial-contaminated industrial surfaces.

Control of spoilage microorganisms in minced pork by a self-developed modified atmosphere induced by the respiratory activity of meat microflora.

The changes in microbial flora of minced pork during aerobic storage at 0, 5, 10 and 15 degrees C were studied. Minced pork samples (100g) were packed using two types of packaging films: (a) a common food film with high permeability (HPF) and (b) a film with low permeability (LPF). The respiratory activity of meat microflora and the use of a LPF resulted in a modified atmosphere in the package headspace developed during storage. Oxygen concentration decreased from 18.7% (after packaging) to 7% (after 15 days of storage) in packages with LPF, stored at 0 degrees C, while CO(2) increased from 3% to 10.5%, respectively. On the contrary, no significant atmosphere changes were observed during storage of HPF packages. The self-developed modified atmosphere in LPF packages resulted in a significant inhibition of pseudomonad growth which was more pronounced at low storage temperatures. For example, during storage at 0 degrees C, the growth rate of pseudomonads in meat packed with LPF was reduced by 48.7% compared to HPF. At 10 degrees C the latter reduction decreased to 13.7%. LPF packaging was also found to inhibit the growth of Brochothrix thermosphacta but this inhibition was weaker compared to pseudomonads. The effect of storage temperature on the growth rate of pseudomonads and B. thermosphacta in minced pork packed with the different films was modeled using an Arrhenius equation. For both bacteria, the activation energy was higher for LPF packaging. This can be attributed to the increased inhibitory effect of the modified atmosphere at lower storage temperature. The Arrhenius model was further used to evaluate the effect of temperature on the time required by the two bacteria to reach a spoilage level of 10(7)CFU/g. The results showed that when LPF packaging is combined with effective temperature control the time-to-spoilage can be significantly extended compared to HPF packaging.

Use of Fourier transform infrared spectroscopy for monitoring the shelf life of ham slices packed with probiotic supplemented edible films after treatment with high pressure processing.

The aim of the present study was to investigate the potential use of Fourier transform infrared (FTIR) spectroscopy to quantify biochemical changes occurring in ham slices packed with probiotic supplemented edible films and treated with High Pressure Processing (HPP), in monitoring spoilage. Details regarding the data collection and experimental procedure were presented by Pavli et al. (2017). A series of Partial Least Squares (PLS) models were developed to correlate spectral data from FTIR analysis with ham spoilage during storage under vacuum at different temperatures (4, 8 and 12°C). FTIR spectra were collected from the surface of the ham samples in parallel with microbiological analysis of total viable counts (TVC) and lactic acid bacteria (LAB). Qualitative interpretation of spectral data was based on a sensory evaluation, using a hedonic scale, classifying the samples in three quality classes, fresh, semi-fresh and spoiled. The scope of the modeling approach was to discriminate the ham slices in their respective quality class and additionally to predict the microbial population directly from spectral data. The results obtained demonstrated that the processing of the samples affected the performance of classification in the sensory classes, with better results observed in the case of for ham slices packed with probiotic supplemented (PS) edible films and of control samples without HPP. The performance of PLS regression models on providing quantitative estimations of microbial counts were based on specific figures of merit (bias factor, accuracy factor, root mean square error, percentage of prediction error). Bias and accuracy factors were close to unity for both microbial groups tested for samples without HPP, whereas for HPP treated samples the values of these indices ranged from 0.963 to 1.332, depending on the case and indice. The results of this study demonstrated for the first time that although FTIR can be used reliably for the rapid assessment of sliced ham, additional processes such as HPP can affect its performance.

Modelling fungal growth using radial basis function neural networks: the case of the ascomycetous fungus Monascus ruber van Tieghem.

A radial basis function (RBF) neural network was developed and evaluated against a quadratic response surface model to predict the maximum specific growth rate of the ascomycetous fungus Monascus ruber in relation to temperature (20-40 degrees C), water activity (0.937-0.970) and pH (3.5-5.0), based on the data of Panagou et al. [Panagou, E.Z., Skandamis, P.N., Nychas, G.-J.E., 2003. Modelling the combined effect of temperature, pH and aw on the growth rate of M. ruber, a heat-resistant fungus isolated from green table olives. J. Appl. Microbiol. 94, 146-156]. Both RBF network and polynomial model were compared against the experimental data using five statistical indices namely, coefficient of determination (R(2)), root mean square error (RMSE), standard error of prediction (SEP), bias (B(f)) and accuracy (A(f)) factors. Graphical plots were also used for model comparison. For training data set the RBF network predictions outperformed the classical statistical model, whereas in the case of test data set the network gave reasonably good predictions, considering its performance for unseen data. Sensitivity analysis showed that from the three environmental factors the most influential on fungal growth was temperature, followed by water activity and pH to a lesser extend. Neural networks offer an alternative and powerful technique to model microbial kinetic parameters and could thus become an additional tool in predictive mycology.

Survival and transfer efficacy of mixed strain Salmonella enterica ser. Typhimurium from beef burgers to abiotic surfaces and determination of individual strain contribution.

The aim of the study was to evaluate the survival and transfer efficacy of 3 Salmonella Typhimurium strains from beef burgers to abiotic surfaces and determine the individual strain distribution. S. Typhimurium population on beef burgers during incubation remained constant at initial levels of contamination approximately 3 and 5 log CFU/g. Additionally, the survival of pathogens on soiled HDPE surfaces was significant during incubation at both initial inocula, while ca 1.5 log CFU/cm2 reduction was observed at 168h. The log transformed transfer rate (log10Tr) was -1.86±0.23 and -1.75±0.40 for high and low inoculum. The level of initial contamination did not have any statistical important impact on bacterial transfer (P>0.05). In addition, the results regarding the strain contribution revealed rather random individual proportion of each strain, recovered from HDPE, SS surfaces and beef burgers. However, the dominance of each strain was strongly dependent on surface at low inoculum and time in case of high inoculum. This observed strain variability during survival and transfer of S. Typhimurium might be of great importance in order to understand and consequently limit the possibility of cross contamination during food processing in a common household.

Effect of high-barrier packaging films with different oxygen transmission rates on the growth of Lactobacillus sp. on meat fillets.

The goal of this study was to determine the combined effect of (i) the oxygen transmission rate (OTR) of packaging film, often called oxygen film permeability or film permeability and (ii) temperature on the growth rate of the main prevailing organism, Lactobacillus sp., in 100% CO2-packed sterile meat fillets. Multifactorial experiments were designed to study the effect of OTR and temperature (0, 5, 8, and 10 degrees C) on the growth rate of Lactobacillus sp. inoculated on sterile meat fillets under 100% CO2 and aerobic conditions. The packaging conditions (air or 100% CO2) and the film OTR significantly affected the growth rate of Lactobacillus sp. only at temperatures higher than 0 degrees C. Low-permeable films with different OTRs did not affect the final population of the bacterium, but the growth rate was significantly changed. The correlation of an ephemeral microbial association with a low spoilage potential (e.g., lactic acid bacteria) or their growth retardation cannot always be assumed unless other determinants (e.g., OTR) of equal importance are taken into account. The present study provides information that can be of benefit to industry and the consumer.

Transfer of Salmonella enterica Serovar Typhimurium from Beef to Tomato through Kitchen Equipment and the Efficacy of Intermediate Decontamination Procedures.

It is well established that a high percentage of foodborne illness is caused by failure of consumers to prepare food in a hygienic manner. Indeed, a common practice in households is to use the same kitchen equipment for both raw meat and fresh produce. Such a practice may lead to cross-contamination of fruits and vegetables, which are mainly consumed without further processing, with pathogenic microorganisms originating from raw meat. The present study was performed to examine the transfer of the pathogenic bacterium Salmonella enterica serovar Typhimurium from inoculated beef fillets to tomatoes via contact with high-density polyethylene (PE), stainless steel (SS), and wooden (WD) surfaces and through cutting with SS knives. Furthermore, the following decontamination procedures were applied: (i) rinsing with tap water, (ii) scrubbing with tap water and liquid dish detergent, and (iii) using a commercial antibacterial spray. When surfaces and knives that came into contact with contaminated beef fillets were not cleaned prior to handling tomatoes, the lowest level of pathogen transfer to tomatoes was observed through PE surfaces. All of the decontamination procedures applied were more effective on knives than on surfaces, while among the surface materials tested, WD surfaces were the most difficult to decontaminate, followed by PE and SS surfaces. Mechanical cleaning with tap water and detergent was more efficient in decontaminating WD surfaces than using commercial disinfectant spray, followed by rinsing only with water. Specifically, reductions of 2.07 and 1.09 log CFU/cm(2) were achieved by washing the WD surfaces with water and detergent and spraying the surfaces with an antibacterial product, respectively. Although the pathogen's populations on SS and PE surfaces, as well as on tomatoes, after both aforementioned treatments were under the detection limit, the surfaces were all positive after enrichment, and thus, the potential risk of cross-contamination cannot be overlooked. As demonstrated by the results of this study, washing or disinfection of kitchen equipment may not be sufficient to avoid cross-contamination of ready-to-eat foods with foodborne pathogens, depending on the decontamination treatment applied and the material of the surfaces treated. Therefore, separate cutting boards and knives should be used for processing raw meat and preparing ready-to-eat foods in order to enhance food safety.

Development of a Safety Monitoring and Assurance System for chilled food products.

The principles of a novel chill chain management policy, coded Safety Monitoring and Assurance System (SMAS) for the optimisation of the distribution of chilled food products within the chill chain are developed. In this system, a new approach based on actual risk evaluation at important points of the chill chain is used in order to promote products to the next stage of distribution. This evaluation based on product's time-temperature history, variation in product's characteristics (e.g. a(w), pH, etc.), and the use of predictive models for the growth of food pathogens, allows to give priority to products in such a way that risk at consumption time is minimized. The effectiveness of SMAS was evaluated against the First In First Out (FIFO) approach, the current method for food distribution, in a case study on the risk of listeriosis of cooked ham using the Monte Carlo simulation technique. Furthermore, the two approaches were compared for their effect on the quality of the products in terms of remaining shelf life at the time of consumption. The results showed that following the SMAS approach the risk of listerisosis is significantly lower while the spoiled products at the time of consumption are significantly reduced compared to FIFO approach.

Field evaluation of the application of time temperature integrators for monitoring fish quality in the chill chain.

The applicability of time temperature integrators (TTI) as effective tools of chill chain monitoring was assessed. Validated kinetic models of pseudomonads growth of Mediterranean, marine-cultured chilled gilt-head seabream (Sparus aurata) and full knowledge of the response of suitable enzymatic TTI are the basis of the TTI application algorithm. This scheme was evaluated through a controlled field test of exported fish, from harvest to final consumption. Response of TTI attached on different locations of packages was compared to actual temperature recording. Data that could not be obtained during the actual field test, such as microbiological or sensory tests of fish at intermediate points of the chain, were measured in a replicate laboratory study, simulating the handling of products and the real time-temperature profiles of the field test. The conducted field tests showed the applicability and usefulness of TTI monitoring of the fish chill chain, elucidating also the practical difficulties and limitations, that need to be addressed for expanding TTI use as a reliable management tool.

RP-HPLC analysis of the phenolic compounds of plant extracts. investigation of their antioxidant capacity and antimicrobial activity.

Extracts of aromatic plants of Greek origin were examined as potential sources of phenolic compounds. RP-HPLC with UV detection was employed for the identification and quantification of the phenolic antioxidants, present in methanolic extracts. The most abundant phenolic acids were ferulic acid (1.1-280 mg/100 g of dry sample) and caffeic acid (1.2-60 mg/100 g of dry sample). (+)-Catechin and quercetin were the most abundant flavonoids. Apigenin and luteolin were detected in high amounts in Menta pulegium and Thymus vulgaris, respectively. The antioxidant capacity was determined, in dried ground plants and in their methanol extracts, with the Rancimat test using sunflower oil as substrate. Both pulverized plants and extracts showed antioxidant capacity. Total phenolic content in the extracts was determined spectrometrically according to the Folin-Ciocalteu assay and ranged from 1 to 21 mg of gallic acid/100 g of dry sample. Antimicrobial activity of the extracts against selected microbes was also conducted in this study.

Effect of temperature, pH, and water activity on biofilm formation by Salmonella enterica enteritidis PT4 on stainless steel surfaces as indicated by the bead vortexing method and conductance measurements.

An assay was developed in an effort to elucidate the effect of important environmental parameters (temperature, pH, and water activity [aw]) on Salmonella Enteritidis biofilm formation on stainless steel surfaces. To achieve this, a modified microbiological technique used for biofilm studying (the bead vortexing method) and a rapid method based on conductivity measurements were used. The ability of the microorganism to generate biofilm on the stainless surfaces was studied at three temperatures (5, 20, and 37 degrees C), four pH values (4.5, 5.5, 6.5, and 7.4), and four aw values (0.5, 1.5, 5.5, and 10.5% NaCl). Results obtained by the bead vortexing method show that maximum numbers of adherent bacteria per square centimeter (106 CFU/cm2) were attained in 6 days at 20 degrees C. Biofilm formation after 7 days of incubation at 20 degrees C was found to be independent of the pH value. In addition, the high concentration of sodium chloride (10.5% NaCl, aw = 0.94) clearly inhibited the adherence of cells to the coupons. Conductance measurements were used as a supplementary tool to measure indirectly the attachment and biofilm formation of bacterial cells on stainless steel surfaces via their metabolic activity (i.e., changes in the conductance of the growth medium due to microbial growth or metabolism). Results obtained by conductance measurements corresponded well to those of the bead vortexing method. Furthermore, we were able to detect cells that remained attached on the metal surfaces even after vortexing via their metabolic activity. The results, except for demonstrating environmental-dependent Salmonella Enteritidis biofilm formation, indicated that traditional vortexing with beads did not remove completely biofilm cells from stainless steel; hence, conductance measurements seem to provide a more sensitive test capable to detect down to one single viable organism.

Performance of Pseudomonas CFC-selective medium in the fish storage ecosystems.

Pseudomonas agar base supplemented with cephaloridine, fucidin, and cetrimide (CFC) was used to count Pseudomonas populations on fish. Both Enterobacteriaceae and Shewanella putrefaciens were able to grow on the CFC medium. Evaluation of the performance of CFC-selective for pseudomonads medium, on fish samples stored aerobically and under a modified atmosphere at 0, 10 and 20 degrees C was tested. The selectivity of the medium was affected by storage temperatures and the type of packaging of the fish samples. The selectivity of the medium diminished as the population increased and for samples stored at high temperature (20 degrees C) or under modified atmospheres. When designing adequate selectivity of a medium, interfering organisms should be taken into account, especially when the background flora tends to be more robust than the organisms to be counted or detected.

Application of a systematic experimental procedure to develop a microbial model for rapid fish shelf life predictions.

A systematic experimental procedure for fish shelf-life modelling was used to develop a model for predicting the quality of fish in the chill chain. For this, the growth of the naturally occurring bacteria pseudomonads, Shewanella putrefaciens, Enterobacteriaceae, lactic acid bacteria and yeasts, on gilt-head seabream (Sparus aurata), was studied at temperatures from 0 to 15 degrees C. The results from the microbiological, organoleptical and chemical analysis conducted on naturally contaminated fish as well as on inoculated sterile fish blocks identified pseudomonads as a good spoilage index. Growth of pseudomonads was modelled as a function of storage temperature and correlated to organoleptical shelf life. To reduce the time required for the enumeration of the initial pseudomonads number, which is crucial information for shelf life prediction, a conductance assay was established. Compared with the conventional microbiological tests, this method gave results in one-fourth of the time.