Immunomodulatory effect of non-viable components of probiotic culture stimulated with heat-inactivated Escherichia coli and Bacillus cereus on holoxenic mice.

BACKGROUND: Competition of probiotic bacteria with other species from the intestinal microbiota involves different mechanisms that occur regardless of probiotics' viability. The objective of this paper was to assess the cytokine serum levels in holoxenic mice after oral administration of non-viable components (NVC) of Enterococcus faecium probiotic culture stimulated with heat-inactivated Escherichia coli and Bacillus cereus in comparison to NVC of unstimulated E. faecium probiotic culture. METHODS: Probiotic E. faecium CMGb 16 culture, grown in the presence of heat-inactivated cultures of E. coli and B. cereus CMGB 102, was subsequently separated into supernatant (SN) and heat-inactivated cellular sediment (CS) fractions by centrifugation. Each NVC was orally administered to holoxenic mice (balb C mouse strain), in three doses, given at 24 hours. Blood samples were collected from the retinal artery, at 7, 14, and 21 days after the first administration of the NVC. The serum concentrations of IL-12 and tumor necrosis factor-alpha (TNF-α) interleukins were assessed by ELISA method. RESULTS: After the oral administration of SN component obtained from the probiotic culture stimulated with heat-inactivated cultures of B. cereus CMGB 102 and E. coli O28, the serum concentrations of IL-12 were maintained higher in the samples collected at 7 and 14 days post-administration. No specific TNF-α profile could be established, depending on stimulated or non-stimulated probiotic culture, NVC fraction, or harvesting time. CONCLUSION: The obtained results demonstrate that non-viable fractions of probiotic bacteria, stimulated by other bacterial species, could induce immunostimulatory effects mediated by cytokines and act, therefore, as immunological adjuvants.

Modulation of bacterial translocation in mice mediated through lactose and human milk oligosaccharides.

Massive resection of the small intestine in infants is imposed to the regulation of several intestinal pathological situations, as intestinal adaptation cannot be relied upon. Many nutritional disturbances are occurring following surgery procedure. In this vein, long-term parenteral feeding is adopt to improve prognosis not always successfully. Clostridia and more specifically Clostridium perfringens, are suspected to participate in the physiopathology of the rising situation. In order to investigate the effect of lactose and human milk neutral oligosaccharides (HMNOs) on Clostridia, germfree mice were inoculated either with enterotoxigenic C.perfringens strain isolated from a patient with NEC, or with a human microbiota harboring C.clostridioforme group(HF). In this vein, different doses of lactose were administrated during 2 weeks in adult mice on an attempt to evaluate the lactase activity. Intake of lactose (70 g/L) and HMNOs (7 g/L) in C.perfringens monoassociated mice induced mortality within a week. In HF mice, no mortality was observed. An increase in Clostridia occurrence was observed in the median ileum after intake of 7 g lactose (p = 0.017). Higher clostridial numbers occurred in caecum following intake of 70 g lactose (p < 0.05) and HMNOs (p < 0.025). Bifidobacteria were found increased from distal ileum to colon following 70 g of lactose intake, whereas they decreased in the caecum of mice drinking lower lactose concentrations. Finally, bacteremia was more frequent in 70 g lactose/L mice (p < 0.02), whereas at lower doses of lactose bifidobacterial translocation was observed. As a result, human milk oligosaccharides could favor clostridial population when reaching the lower intestine. The shortness of the small intestine in infants underwent massive intestinal resection seems to be associated to an incomplete breakdown of lactose. Enteral feeds formulas deprived in lactose would be more suitable in enteral feeding of infants.

Selective decontamination by antimicrobials during long term treatment: perspectives for saving host indigenous microbiota.

Short bowel syndrome (SBS) occurs in patients fed total parenteral nutrition (TPN) after massive intestinal resection. TPN weaning is often associated with occlusion or sepsis. In the present study the intestinal biotope was investigated in young patients (n = 14) with massive intestinal resection and recurrent symptoms of sepsis or occlusion during enteral food introduction. They were treated by aminosides for a long term period. Ileal effluents were collected for enumerating bacteria. In some case, blood and rectal specimen were also collected. A few patients developed bacterial overgrowth (1), occlusion (1), sepsis (4), osteoarthritis (1) or pneumonia (1) during the survey. A drastic drop of bifidobacteria that was not prevented by human milk feeding was observed prior occlusion or respiratory infection. Detection of clostridial vegetative forms preceded sepsis and decrease in clostridia parallelled recovery. In conclusion, onset of symptoms was related with extreme imbalance of the ileal flora. Supplementation with bifidobacterial compounds that were well tolerated in two patients could be of interest in children with recurrent symptoms.

Antibiotic activity of tigecycline against clinical pathogens by the micro dilution method.

Resistant pathogens are the cause of clinical infections which threatening the patients lives and challenging the health systems through their economic importance. Therefore, new antibacterial agents with a broader spectrum of activity that protect against development of resistance are required. Tigecycline (Tygacil, Wyeth) is a relatively new FDA and EMEA approved glycylcycline antimicrobial with an expanded broad-spectrum activity against pathogens involved in complicated skin and skin structure infections. In this study we evaluated the in vitro activity of tigecycline in comparison to 14 other antibiotics against 182 clinical pathogens by use of the micro dilution method. In overall, tigecycline exhibited the lowest Minimum Inhibitory Concentration (MIC) values in almost all bacteria with a mean of 0.52 ± 1.25mg/L, followed by meropenem and levofloxacin (mean MIC values 1.29 ± 2.52 and 1.45 ± 3.078 mg/L, respectively). MIC50 and MIC90 values of tigecycline were: 0.06 and 0.15 mg/L for E. coli, 0.12 and 1.00 mg/L for Klebsiella sp., 0.12 and 0.85 mg/L for various Enterobacter sp., 1.00 and 8.00 mg/L for Pseudomonas sp., 0.25 and 1.00 mg/L for Acinetobacter sp., 0.06 and 0.12 mg/L for Serratia sp., 0.12 and 0.25mg/L for Staphylococcus aureus, 0.5 and 5.00 mg/L for Streptococcus sp. The MIC values recorded were among the lowest in recent literature for Acinetobacter sp. (included A. baumannii), and comparable to those obtained for Klebsiella, Serratia and Enterobacter indicating that tigecycline has a promising in vitro activity.

Microbial ecology and quality assurance in food fermentation systems. The case of kefir grains application.

Fermentation technology has become a modern method for food production the last decades as a process for enhancing product stability, safety and sensory standards. The main reason for this development is the increasing consumers' demand for safe and high quality food products. The above has led the scientific community to the thorough study for the appropriate selection of specific microorganisms with desirable properties such as bacteriocin production, and probiotic properties. The main food products produced through fermentation activity are bread, wine, beer cheese and other dairy products. The microorganisms conducting the above processes are mainly yeasts and lactic acid bacteria. The end products of carbohydrate catabolism by these microorganisms contribute not only to preservation as it was believed years ago, but also to the flavour, aroma and texture and to the increase of the nutritional quality by thereby helping determine unique product characteristics. Thus, controlling the function of specific microorganisms or the succession of microorganisms that dominate the microflora is therefore advantageous, because it can increase product quality, functionality and value. Throughout the process of the discovery of microbiological diversity in various fermented food systems, the development of starter culture technology has gained more scientific attention, and it could be used for the control of the manufacturing operation, and management of product quality. In the frame of this review the presentation of the quality enhancement of most consumed fermented food products around the world is attempted and the new trends in production of fermented food products, such as bread is discussed. The review is focused in kefir grains application in bread production.

Application of novel starter cultures for sourdough bread production.

Sourdough application has been extensively increased in the last years due to the consumers demand for food consumption without the addition of chemical preservatives. Several starter cultures have been applied in sourdough bread making targeting the increase of bread self-life and the improvement of sensorial character. More specific, Lactobacillus acidophilus and Lactobacillus sakei as single and mixed cultures were used for sourdough bread making. Various sourdough breads were produced with the addition of sourdough perviously prepared with 10% w/w L. acidophilus, 10% w/w L. sakei and 5% w/w L. acidophilus and 5% w/w L. sakei at the same time. Various chemical parameters were determined such as lactic acid, total titratable acidity and pH. The results revealed that the produced sourdough bread made with sourdough containing the mixed culture was preserved for more days (12 days) than all the other breads produced in the frame of this study, since it contained lactic acid in higher concentrations. The respective total titratable acidity varied between 10.5 and 11 ml NaOH N/10. The same sourdough bread had a firmer texture, better aroma, flavor and overall quality compared to other sourdough breads examined in this study, as shown by sensory evaluation tests and results obtained through SPME GC-MS analysis, which revealed significant differences among the different bread types.

Microbiological quality and related factors of sheep milk produced in farms of NE Greece.

Sheep herds are a significant capital of dairy industry in Greece, but as dairy farming becomes more complex and intense the need to provide assistance to dairy producers on milk quality is critical. In this study, the quality of raw milk produced in sheep farms of NE Greece was surveyed and the role of various factors was explored. In total, 21 dairy ewe's farms from the regions of Xanthi and Evros, in the north-eastern Greece were monitored. Milk samples were sampled after the morning milking every 15 days throughout the dairy period (March-June). For the study, a questionnaire was also filled by personal interview with the owners in order to collect information about herd characteristics, health status, handling practices etc. From each farm, air was also sampled for microbiological analysis. Milk samples were examined for chemical components: fat content, protein, lactose, non-fat dry matter (NFDM) and somatic cells count (SCC). Microbiological examination involved the estimation of Total Bacterial Counts (TBC), coliform count (CC), Staphylococcus aureus, Streptococcus sp., and preliminary incubation count (PIC). The possible correlation among different bacterial species and their interaction with SCC and chemical components of milk was also considered. It was examined whether farm management practices could influence the hygiene and the quality of milk. Our results show that as an average TBCs were 5.48 log cfu/ml, SCC: 6.05 log cells/ml, CC: 4.49 log cfu/ml, S. aureus: 3.94 log cfu/ml, Streptococcus sp.: 4.95 log cfu/ml and PIC: 5.7 log cfu/ml. The mean fat, protein, lactose and NFDM were 6.17%, 5.28%, 4.73% and 10.95% respectively. The study revealed significant positive correlation between TBC and PIC (0.825), while SCC was marginally positive correlated with protein and NFDM. No statistically significant correlations observed among SCC with any of the bacterial species. Herd size and farm management practices had considerable influence on SCC and bacterial species.

Microbial ecology of the watery ecosystems of Evros river in North Eastern Greece and its influence upon the cultivated soil ecosystem.

The aim of the present study was to evaluate the microbial ecosystem of cultivated soils along the Evros river in NE Greece. Evros river together with its derivative rivers constitute the capital source of life and sustainable development of the area. Along this riverside watery ecosystem systematic agro-cultures were developed such as wheat, corn and vegetable cultures. The evaluation of the ecosystem microbial charge was conducted in both axes which are the watery ecosystem and the riverside cultivated soil area. Considerable discrimination of water quality was observed when considering chemical and microbiological parameters of the Evros river ecosystem. Ardas river possesses a better water quality than Evros and Erythropotamos, which is mainly due to the higher quantities that these two rivers accumulate from industrial, farming and urban residues leading to higher degree of pollution. An increased microbial pollution was recorded in two of the three rivers monitored and a direct relation in microbial and chemical charging between water and cultivated-soil ecosystems was observed. The protection of these ecosystems with appropriate cultivated practices and control of human and animal activities will define the homeostasis of the environmental area.

Hygienic quality and antibiotic resistance profile of sliced butchery.

In order to investigate the microbiological quality of different meat products on the Greek market, 200 samples were collected from the following preparations: boiled turkey (n=50), boiled pork ham (n=50), smoked turkey (n=50) and smoked pork ham (n=50). In all cold meat preparations Clostridium perfringens vegetative and spore forms, Staphylococcus aureus, Escherichia coli and other Clostridium sp lec(-), as well as Lactobacillus, Bacillus sp. and Salmonella sp. were recovered. For instance Bacillus cereus was present in 6% of the samples. L. monocytogenes, Campylobacter jejuni and Campylobacter coli were rarely present (1-4%) while Yersinia enterocolitica and Campylobacter lari were absent. Differences in the occurrence of S. aureus, Salmonella sp., E. coli and spore forms of C. perfringens in boiled and smoked samples, reflects either the differences in the processing of the foods or could be associated to the extensive handling by the personnel during the purchasing (storage, slicing, wrapping). Antibiotic resistance on specific antibiotics for each pathogen was also studied. A multiresistance antibiotic profile was effective for most bacterial strains, and pronounced resistance profiles were observed for the commonly used antibiotics as ampicillin, penicillin, cephalothin, streptomycin followed by ceftriaxone and gentamycin. Albeit this high observed resistance profile, the tested strains generally conserved their susceptibility to amikacin, aztreonam, chloramphenicol and tylosin conserved an almost absent resistance. Antibiotics commonly used for therapeutic purposes, as well as antibiotics added to feed stuff of animals for increasing animal flesh production should contribute to the extensive spreading of antibiotic resistance in food and the environment. Systematically monitoring of the microbiological quality of cold butchery preparations must be done, in order to preserve food quality, optimizing the processing and elaboration methods of the product and safeguard the public health.

Antibacterial activity of different honeys against pathogenic bacteria.

To study the antimicrobial activity of honey, 60 samples of various botanical origin were evaluated for their antimicrobial activities against 16 clinical pathogens and their respective reference strains. The microbiological quality of honeys and the antibiotic susceptibility of the various isolates were also examined. The bioassay applied for determining the antimicrobial effect employs the well-agar diffusion method and the estimation of minimum active dilution which produces a 1mm diameter inhibition zone. All honey samples, despite their origin (coniferous, citrus, thyme or polyfloral), showed antibacterial activity against the pathogenic and their respective reference strains at variable levels. Coniferous and thyme honeys showed the highest activity with an average minimum dilution of 17.4 and 19.2% (w/v) followed by citrus and polyfloral honeys with 20.8 and 23.8% respectively. Clinical isolates of Staphylococcus aureus subsp. aureus, Escherichia coli, Salmonella enterica subsp. Enterica, Streptococcus pyogenes, Bacillus cereus and Bacillus subtilis were proven to be up to 60% more resistant than their equal reference strains thus emphasizing the variability in the antibacterial effect of honey and the need for further research.

Microbiological quality of grey-mullet roe.

The Greek grey-mullet roe is produced from the fully developed gonads of the female mullet (Mugil cephalus) couth in lagoons during their reproductive migration. The traditional processing method of the roe includes, air drying, salting, shape formation and covering with multiple layers of natural beeswax for preservation and distribution. Fish Roe brands have been a staple in local diet and is increasingly becoming popular in the international market. As a ready-to-eat food it's microbial quality should be of concern for the protection of consumers health. In this study, 48 samples of fish roe, just before waxing, were collected from various local processors for microbiological examination by using selective media and incubated under aerobic and anaerobic conditions. The identification of the bacteria was carried out according to the Bergey's manual. Microscopic examination of Gram stained cells, catalase, oxidase and biochemical tests were performed when necessary to further identify. V. parahaemolyticus, Vibrio spp., Salmonella spp., and Aeromonas hydrophila were detected in one sample (2%). Shigella spp., and Flavobacterium spp. in two samples (4%), Clotriduim perfringens (vegetative forms), E. coli, and spores of Bacillus spp., were detected in three samples (6%), Staphylococcus aureus in four samples (8%). Various Micrococcus spp., and spores of C. perfringens in 16% and 35% of the samples respectively. From the Listeria genus, only the species Listeria innocua, Listeria welshimeri, Listeria seeligeri Listeria ivanovii and Listeria grayi were recovered from 2 to 10% of the samples. Microbiological analyses revealed the presence of a small number of pathogens in grey-mullet roe samples which are in accordance with the findings of similar studies. Traditional processing of the fish roe, seems inadequate to ensure the food safety and even waxing isn't expected to fully protect them against facultative anaerobes with salt tolerance. Therefore, additional measures should be taken during processing and marketing of fish roe to minimize potential health risks for the consumers.

Antibacterial activities of essential oils from eight Greek aromatic plants against clinical isolates of Staphylococcus aureus.

Aromatic plants have been used widely to extend the shelf life of foods but at the same time research is undergoes for their properties as antibacterial agents in clinical use. Although there are promising results for the antimicrobial properties of various essential oils against environmental or food-isolated strains of Staphylococcus aureus, limited work has been done concerning these properties against clinical isolates of this pathogen. S. aureus is responsible for an increase number of nosocomial infections and at the same time exhibits increased resistance to synthetic agents. In this study, essential oils from eight aromatic plants common in Greece were isolated by hydrodistillation, analyzed by gas chromatography (GC) and GC/mass spectrometry (GC/MS) for their chemical components and tested for their antimicrobial activities against 24 clinical isolates of S. aureus. The methods used were disk diffusion and broth dilution in order to determine the Minimum Inhibitory Concentration (MIC). Our results showed that essential oils from Origanum vulgare and Origanum dictamnus were active against S. aureus when tested by disk diffusion, but exhibited increased MIC values (>256 mg/L) with the dilution method. In contrast, the reference strain NCTC 6571 showed to be extremely sensitive in most of the oils tested (MICs 0.25-32.0 mg/L) and resistant only to the essential oil from Ocimum basilicum. Therefore, there is no evidence of a potential clinical use for those essential oils and further research is needed in order to determine if they could substitute efficiently synthetic antibiotics or, perhaps be used in combination.

Microbial ecology of fish species ongrowing in Greek sea farms and their watery environment.

The present study focuses on the bacteriological profile of both watery ecosystem and fishes from different North and Central West Greek fish-farms ongrowing euryhaline fish species. The natural microflora of the fish and the water of their ongrowing units in selected farms were studied for a period of 20 months. The analyzed samples were mainly sea bream (Sparus aurata) 61.3% and sea bass (Dicentrarchus labrax) 24%. In most of the watery ecosystems coming from the different sampling areas, total and fecal coliforms as well as total and fecal streptococci were abundant in all water samples. Enterococcus, Escherichia coli and Pseudomonas were present at a level of 3 logs cfu/100 ml. The anaerobic Clostridium perfringens was found in vegetative (21.3%) and spore forms (13.3%). It is of interest to note that pathogens as Pasteurella piscicida and Vibrio anguillarum were isolated only in a small number of samples. Staphylococcus aureus was detected in 4% of the samples, other Staphylococcus sp. in 29.3%, E. coli in 30.7%, Salmonella sp. in 1.3%, Pseudomonas sp. in 13.3%, Clostridia lec(-) in 49.3%, Bacillus sp. in 38.7%, Vibrio sp. in 18.7%, Lactobacillus and Lactococcus sp. in 36% και 29.3% respectively. Vegetative forms of C. perfringens were detected in 22.7%. Although, our results showed no significant correlations between the sea water and fish microflora, more focus on this bipolar interacting system should be necessary in order to avoid any possible disturbance in the balance of the healthy farming ecosystem with the host organisms.

Microbial challenges of poultry meat production.

Food safety and shelf-life are both important microbial concerns in relation to broiler meat production. Focus is mainly placed on the absence or control of potentially pathogenic microbes such as Salmonella spp. and Campylobacter spp. but, from the commercial point of view, other spoilage bacteria also play a role as potential threats. Regarding food safety, the primary target should be the production of pathogen-free live animals, thus allowing slaughter plants to keep the processing line free of those microorganisms. Consumers believe that quality of foods from organic production is superior to foods from conventional production. The aim of the present study was to evaluate and compare the bacterial quality of chicken meat from organic and conventional production on the basis of traditional meat quality criteria. Fresh free grazing broiler carcasses were purchased directly from rural households (n = 80) and fresh retail chicken parts from conventional broiler carcasses from the local supermarkets in the region of Epirus (Poultry Producers Association. Arta) (n = 200). The samples were microbiologically tested for the presence of bacteria such as: Salmonella spp., Listeria monocytogenes, Staphylococcus aureus, Enterobacteriaceae, Escherichia coli, Campylobacter spp., and C. perfringens. Total count of aerobic mesophilic bacteria was also determined. Bacteriological tests were performed by means of standard methods of isolation and identification of individual species of bacteria according to ISO requirements. API-tests (bioMerieux) and Vitek 2 Identification System (bioMerieux) were used for biochemical determination. High levels of microbial contamination and occurrence of pathogenic bacteria at then fresh free grazing broiler carcasses reflect the poor hygienic quality of the slaughter conditions in the rural households.

Isolation of microbial pathogens of subclinical mastitis from raw sheep's milk of Epirus (Greece) and their role in its hygiene.

The natural raw milk microflora is a factor that expresses its sensorial characteristics. The microbial charge into the mammary gland of healthy animal is low and the application of right and healthy conditions during milking and cheese making procedure, prevents from contaminating as well as maintains the natural microflora in order to lend the particular characteristics of milk. The purpose of the present project was the study of the Total Viable Count (T.V.C.) and the count of total psychrotropic bacteria of raw sheep milk from Boutsiko and Karamaniko breeds, collected from healthy animals, as well as the isolation, identification and enumeration of pathogenic bacteria related with the hygiene and the quality of raw sheep milk (with a particular interest in bacteria that may cause human infection). During the experiment we examined two hundred forty (240) samples of raw sheep milk. In these samples a) Staphylococcus aureus, Salmonella sp., Escherichia coli, Clostridium perfringens (vegetative cells and spores) and Bacillus sp. were isolated and identified b) the Total Viable Count and the total number of psychrotropic bacteria were also specified. The sampling, the preparation of samples and decimal dilutions were based on international methods. The Total viable count was determined using the standard methods of the American Public Health Association, 2002. The total number of psychrotropic bacteria was determined using APHA 1976, 1978 rules. The identification of the bacteria was carried out according to the Bergey's manual. Microscopic examination of Gram stained cells, catalase, oxidase and biochemical tests were performed when necessary to further identify. From the 240 milk samples tested, only 5% were E. coli positive, with mean counts ranged from 2 × 10(3) to 2.4 × 10(4) cfu/ml. S. aureus was isolated from 24% of the samples and the mean count per ml was ranged from <10 to 3.4 × 10(2). Meanwhile, Bacillus spp. was also detected in 29% samples. Vegetative forms and spores of C. perfringens were detected in 13% and 63% of the samples respectively. However, microbiological analyses revealed the presence of a small number of selected pathogens in milk samples such as Salmonella, which was only detected in 5% of the samples. Listeria sp., Pseudomonas sp. and Vibrio cholerae were never found. From the experimental results, the Total Viable Count from raw sheep milk samples, fulfils the microbiological criteria of EU Legislation in a percentage of approximately 97%.

Occurrence of Clostridium perfringens from different cultivated soils.

The occurrence of Clostridium perfringens was estimated in 750 samples originated from a variety of soils bearing various bulb crops: Brawnica oderacea (vegetable), Olea europaea, Daucus carota (carote), Solanum tuberosum (potato), Phaseolus vulgaris (green haricot), Beta vulgaris var. rapaceum (beetroot), Cucurbita pepo (squash), Allium cepa (onion), Cucumis sativus (cucumber) and Capsicum annum (pepper). All isolated strains were tested for their antimicrobial activities to amoxicillin, penicillin G, kanamycin, tetracycline, streptomycin, erythromycin, chloramphenicol and metronidazole. When considering the type of the bulb production, it was observed increased number of C. perfringens spore densities in the most undersurface bulb soils. Moreover, C. perfringens spore are likely to occur in particularly large numbers in soil contaminated by fecal matter. Additionally, there is a close relationship between the spore amount and nature of organic content. Presence of C. perfringens was associated with acidic soil. Most of our strains showed resistance to the studied antibiotics applied usually for human and veterinary care. A systematic monitoring of the cultivated soil ecosystems must include bacteriological parameters together with chemical indices of organic pollution in order to obtain information adequate for assessing their overall quality.

Physical, chemical and microbiological quality of ice used to cool drinks and foods in Greece and its public health implications.

Ice used for direct human consumption or to preserve foods and cool down drinks can be contaminated with pathogenic microorganisms and may potentially become a vehicle for consumer's infection. To evaluate physical, chemical and microbiological quality of commercial ice and ice used for fish and seafood, 100 ice samples collected at 10 different retail points in the region of Epirus were studied. The following microbiological parameters were determined: Total coliforms, fecal coliforms, Salmonella spp., Shigella spp., Yersinia spp., Escherichia coli, Campylobacter sp., Vibrio cholerae, Aeromonas spp., Pseudomonas aeruginosa and Clostridium perfringens. E. coli was detected in 22% and coliforms were detected in 31% of samples. Samples in which coliforms were detected fail to meet the microbiological criteria specified by the drinking water legislation. Aeromonas spp., Shigella spp., Campylobacter sp. and V. cholerae were not detected. Spore forms of C. perfringens were prevalent at 35% and the psychotropic bacterium's P. aeruginosa and Yersinia spp. were found only at three samples each. The presence of large numbers of coliforms as well as of other pathogenic strains suggested that commercial ice and ice used to make cool drinks or in preservation of fish and seafood may represent a potential hazard to the consumer. In view of the results reported herein, it is highly recommended that national regulatory guidelines should be established for the production of ice as long as regular inspections.

Chemical and microbiological characterization of artisan inoculants used for the fermentation of traditional dairy products in Epirus area (Greece).

The artisan production of a variety of dairy products is a historical tradition since antiquity, which is still practiced nowadays, in the area of Epirus, (northwestern Greece). The common denominator of these products, regardless of the final form of the end product, is the fermentation of raw milk by the use of dried artisan inoculants made of the dehydrated rumen of small ruminants, mainly lambs. The aim of this research project is to study the microbiological parameters as well as certain technological parameters (chymosin activity and lipolytic activity) of these inoculants. The results revealed a wide biodiversity of microorganisms such as Lactobacilli, Lactococci, Leuconostoc, Pediococci, Streptococci, Bifidobacteria, Enterococci, Clostridia and coliforms. Chymosin activity and lipolytic activity were found to be higher in artisan inoculants than in the commercial ones.

Predictive Modeling of Microbial Behavior in Food.

Microorganisms can contaminate food, thus causing food spoilage and health risks when the food is consumed. Foods are not sterile; they have a natural flora and a transient flora reflecting their environment. To ensure food is safe, we must destroy these microorganisms or prevent their growth. Recurring hazards due to lapses in the handling, processing, and distribution of foods cannot be solved by obsolete methods and inadequate proposals. They require positive approach and resolution through the pooling of accumulated knowledge. As the industrial domain evolves rapidly and we are faced with pressures to continually improve both products and processes, a considerable competitive advantage can be gained by the introduction of predictive modeling in the food industry. Research and development capital concerns of the industry have been preserved by investigating the plethora of factors able to react on the final product. The presence of microorganisms in foods is critical for the quality of the food. However, microbial behavior is closely related to the properties of food itself such as water activity, pH, storage conditions, temperature, and relative humidity. The effect of these factors together contributing to permitting growth of microorganisms in foods can be predicted by mathematical modeling issued from quantitative studies on microbial populations. The use of predictive models permits us to evaluate shifts in microbial numbers in foods from harvesting to production, thus having a permanent and objective evaluation of the involving parameters. In this vein, predictive microbiology is the study of the microbial behavior in relation to certain environmental conditions, which assure food quality and safety. Microbial responses are evaluated through developed mathematical models, which must be validated for the specific case. As a result, predictive microbiology modeling is a useful tool to be applied for quantitative risk assessment. Herein, we review the predictive models that have been adapted for improvement of the food industry chain through a built virtual prototype of the final product or a process reflecting real-world conditions. It is then expected that predictive models are, nowadays, a useful and valuable tool in research as well as in industrial food conservation processes.

Lactobacillus paracasei K5 displays adhesion, anti-proliferative activity and apoptotic effects in human colon cancer cells.

Lactobacillus paracasei K5 is a lactic acid bacteria (LAB) strain, isolated recently from feta-type cheese. Its probiotic potential has been demonstrated in a series of established in vitro tests. Moreover, incorporation of L. paracasei K5 as starter culture offered organoleptic and technological advantages to novel fermented food products. In the present study, further investigation of the potential probiotic activity of L. paracasei K5 was performed and its mechanisms of action were investigated. Employing quantitative analysis and confocal, fluorescent microscopy the adhesion properties of the above strain were studied. L. paracasei K5 displayed efficient adherence capacity to Caco-2 colon cancer cells, similarly to the reference strains Lactobacillus casei ATCC 393 and Lactobacillus rhamnosus GG. Moreover, treatment of Caco-2 cells with L. paracasei K5 inhibited cell proliferation in a time-and dose-dependent manner. The anti-proliferative effects appear to be mediated through induction of apoptosis via modulation of expression of specific Bcl-2 family proteins. These results elucidate the mechanisms of action of L. paracasei K5 and enhance its potential probiotic activity.