Anti-Salmonella Activity of Thymus serpyllum Essential Oil in Sous Vide Cook-Chill Rabbit Meat.

Food is generally prepared and vacuum-sealed in a water bath, then heated to a precise temperature and circulated in a sous vide machine. Due to its affordability and ease of use, this cooking method is becoming increasingly popular in homes and food service businesses. However, suggestions from manufacturers and chefs for long-term, low-temperature sous vide cooking raise questions about food safety in the media. In this study, heat treatment with different times and wild thyme essential oil (EO) in sous vide-processed rabbit longissimus dorsi muscle were found to inactivate Salmonella enterica. The rabbit meat samples were vacuum-packed in control groups, in the second group the rabbit meat samples were injected with S. enterica, and in the third group were meat samples infected with S. enterica with Thymus serpylum EO additive. The vacuum-packed samples were cooked sous vide for the prescribed time at 55, 60, and 65 °C. At 5, 15, 30, and 60 min, the quantities of S. enterica, total bacterial counts, and coliform bacteria were measured in groups of sous vide rabbit meat. Microbiological analyses of rabbit meat samples on days 1 and 7 were evaluated. In this study, total viable counts, coliforms bacteria, and number of Salmonella spp. were identified. After incubation, isolates from different groups of microorganisms were identified by the mass spectrometry technique. For each day measured, the test group exposed to a temperature of 55 °C for 5 min had a greater number of total microbiota. The most isolated microorganisms by MALDI-TOF MS Biotyper from the control and treated groups were Lactococcus garvieae and in the treated groups also S. enterica. Based on our analysis of sous vide rabbit meat samples, we discovered that adding 1% of thyme essential oil to the mixture reduced the amount of Salmonella cells and increased the overall and coliform bacterial counts. The microbiological quality of sous vide rabbit meat that was kept for seven days was positively impacted by the addition of thyme essential oil.

Thymus zygis, Valuable Antimicrobial (In Vitro and In Situ) and Antibiofilm Agent with Potential Antiproliferative Effects.

With the growing issues of food spoilage, microbial resistance, and high mortality caused by cancer, the aim of this study was to evaluate T. zygis essential oil (TZEO) as a potential solution for these challenges. Here, we first performed GC/MS analysis which showed that the tested TZEO belongs to the linalool chemotype since the abundance of linalool was found to be 38.0%. Antioxidant activity assays showed the superiority of TZEO in neutralizing the ABTS radical cation compared to the DPPH radical. The TZEO was able to neutralize 50% of ABTS•+ at the concentration of 53.03 ± 1.34 µg/mL. Antimicrobial assessment performed by employing disc diffusion and minimal inhibitory concentration assays revealed TZEO as a potent antimicrobial agent with the highest inhibition activity towards tested gram-negative strains. The most sensitive on the treatment with TZEO was Enterobacter aerogenes showing an MIC 50 value of 0.147 ± 0.006 mg/mL and a MIC 90 value of 0.158 ± 0.024 mg/mL. Additionally, an in situ analysis showed great effects of TZEO in inhibiting gram-negative E. coli, P. putida, and E. aerogenes growing on bananas and cucumbers. Treatment with the TZEO vapor phase in the concentration of 500 µg/mL was able to reduce the growth of these bacteria on the food models to the extent > 90%, except for E. coli growth on the cucumber, which was reduced to the extent of 83.87 ± 4.76%. Furthermore, a test on the antibiofilm activity of the tested essential oil revealed its biofilm prevention effects against Salmonella enterica which forms biofilms on plastic and stainless-steel surfaces. Performed tests on the TZEO effects towards cell viability showed no effects on the normal MRC-5 cell line. However, the results of MTT assay of TZEO effects on three cancer cell lines (MDA-MB-231, HCT-116, and K562) suggest that TZEO exerted the strongest effects on the inhibition of the viability of MDA-MB-231 cells, especially after long-term treatment in the highest concentration applied with reducing the viability of the cells to 57%. Additionally, results of NBT and Griess assays suggest that TZEO could be a convenient candidate for future testing for developing novel antitumor therapies.

Dried Herbs as an Easy-to-Use and Cost-Effective Alternative to Essential Oils to Extend the Shelf Life of Sheep Lump Cheese.

The objective of this research was to assess the effectiveness of three specific dried herbs (rosemary, thyme, and oregano) in combating microbial spoilage in sheep lump cheese. This was achieved by comparing them with a control group and cheeses treated with corresponding 1% essential oils (Rosmarinus officinalis, Origanum vulgare, Thymus vulgaris). All cheese samples were vacuum-sealed and stored at 4 °C for 15 days. Analysis of total viable counts of viable bacteria (TVC), coliform bacteria (CB), lactic acid bacteria (LAB), and microscopic filamentous fungi (MFF) was conducted on days 0, 5, 10, and 15. The results revealed that, at the end of the storage period, dried oregano-treated samples exhibited the lowest TVC count (5.80 log CFU/g), while dried rosemary-treated samples showed the lowest CB count (3.27 log CFU/g). Moreover, the lowest MFF count (2.40 log CFU/g) was observed in oregano essential oil-treated samples. Additionally, dried oregano-treated samples displayed the highest LAB count (4.49 log CFU/g) at the experiment's conclusion. Furthermore, microorganism identification from sheep cheese was performed using MALDI-TOF MS Biotyper technology, revealing that the most frequently isolated bacteria were Citrobacter braakii and Hafnia alvei (Enterobacteriaceae family), along with Lacticaseibacillus paracasei (Lactobacillaceae family). In summary, all the natural substances examined exhibited inhibitory effects against the studied microorganisms, with oregano essential oil and dried oregano demonstrating the strongest inhibitory effects. This supports their potential use as cost-effective natural preservatives to extend the shelf life of sheep lump cheese.

Kombucha and Water Kefir Grains Microbiomes' Symbiotic Contribution to Postbiotics Enhancement.

Wild artisanal cultures, such as a symbiotic culture of bacteria and yeasts (SCOBY) and water kefir grains (WKG), represent a complex microorganism consortia that is composed of yeasts and lactic and acetic acid bacteria, with large strains of diversity and abundance. The fermented products (FPs) obtained by the microbiome's contribution can be included in functional products due to their meta-biotics (pre-, pro-, post-, and paraprobiotics) as a result of complex and synergistic associations as well as due to the metabolic functionality. In this study, consortia of both SCOBY and WKG were involved in the co-fermentation of a newly formulated substrate that was further analysed, aiming at increasing the postbiotic composition of the FPs. Plackett-Burman (PBD) and Response Surface Methodology (RSM) techniques were employed for the experimental designs to select and optimise several parameters that have an influence on the lyophilised starter cultures of SCOBY and WKG activity as a multiple inoculum. Tea concentration (1-3%), sugar concentration (5-10%), raisins concentration (3-6%), SCOBY lyophilised culture concentration (0.2-0.5%), WKG lyophilised culture concentration (0.2-0.5%), and fermentation time (5-7 days) were considered the independent variables for mathematical analysis and fermentation conditions' optimisation. Antimicrobial activity against Bacillus subtilis MIUG B1, Staphylococcus aureus ATCC 25923, Escherichia coli ATCC 25922, and Aspergillus niger MIUG M5, antioxidant capacity (DPPH), pH and the total acidity (TA) were evaluated as responses. The rich postbiotic bioactive composition of the FP obtained in optimised biotechnological conditions highlighted the usefulness of the artisanal co-cultures, through their symbiotic metabolic interactions for the improvement of bioactive potential.

Effect of Selected Factors Influencing Biogenic Amines Degradation by Bacillus subtilis Isolated from Food.

Modern food technology research has researched possible approaches to reducing the concentration of biogenic amines in food and thereby enhance and guarantee food safety. Applying adjunct cultures that can metabolise biogenic amines is a potential approach to reach the latter mentioned goal. Therefore, this study aims to study the crucial factors that could determine the decrease in biogenic amines concentration (histamine, tyramine, phenylethylamine, putrescine and cadaverine) in foodstuffs using Bacillus subtilis DEPE IB1 isolated from gouda-type cheese. The combined effects of cultivation temperature (8 °C, 23 °C and 30 °C) and the initial pH of the medium (5.0, 6.0, 7.0 and 8.0) under aerobic and also anaerobic conditions resulted in the decrease of the tested biogenic amines concentration during the cultivation time (another factor tested). Bacillus subtilis was cultivated (in vitro) in a medium supplemented with biogenic amines, and their degradation was detected using the high-performance liquid chromatography equipped with UV-detector. The course of biogenic amines degradation by Bacillus subtilis DEPE IB1 was significantly influenced by cultivation temperature and also the initial pH of the medium (p < 0.05). At the end of the cultivation, the concentration of all of the monitored biogenic amines was significantly reduced by 65-85% (p < 0.05). Therefore, this strain could be used for preventive purposes and contributes to food safety enhance.

Chemical Composition and Biological Activities of Eucalyptus globulus Essential Oil.

Eucalyptus globulus essential oil (EGEO) is considered as a potential source of bioactive compounds with significant biological activity. The aim of this study was to analyze the chemical composition of EGEO, in vitro and in situ antimicrobial activity, antibiofilm activity, antioxidant activity, and insecticidal activity. The chemical composition was identified using gas chromatography (GC) and gas chromatography/mass spectrometry (GC/MS). The main components of EGEO were 1,8-cineole (63.1%), p-cimene (7.7%), a-pinene (7.3%), and a-limonene (6.9%). Up to 99.2% of monoterpenes were present. The antioxidant potential of essential oil and results indicate that 10 µL of this sample can neutralize 55.44 ± 0.99% of ABTS•+, which is equivalent to 3.22 ± 0.01 TEAC. Antimicrobial activity was determined via two methods: disk diffusion and minimum inhibitory concentration. The best antimicrobial activity was shown against C. albicans (14.00 ± 1.00 mm) and microscopic fungi (11.00 ± 0.00 mm-12.33 ± 0.58 mm). The minimum inhibitory concentration showed the best results against C. tropicalis (MIC 50 2.93 µL/mL, MIC 90 3.17 µL/mL). The antibiofilm activity of EGEO against biofilm-forming P. flourescens was also confirmed in this study. The antimicrobial activity in situ, i.e., in the vapor phase, was significantly stronger than in the contact application. Insecticidal activity was also tested and at concentrations of 100%, 50%, and 25%; the EGEO killed 100% of O. lavaterae individuals. EGEO was comprehensively investigated in this study and information regarding the biological activities and chemical composition of the essential oil of Eucalyptus globulus was expanded.

Biological Activity of Cupressus sempervirens Essential Oil.

The aim of this study was to evaluate the antioxidant, antibiofilm, antimicrobial (in situ and in vitro), insecticidal, and antiproliferative activity of Cupressus sempervirens essential oil (CSEO) obtained from the plant leaf. The identification of the constituents contained in CSEO was also intended by using GC and GC/MS analysis. The chemical composition revealed that this sample was dominated by monoterpene hydrocarbons α-pinene, and δ-3-carene. Free radical scavenging ability, performed by using DPPH and ABTS assays, was evaluated as strong. Higher antibacterial efficacy was demonstrated for the agar diffusion method compared to the disk diffusion method. The antifungal activity of CSEO was moderate. When the minimum inhibitory concentrations of filamentous microscopic fungi were determined, we observed the efficacy depending on the concentration used, except for B. cinerea where the efficacy of lower concentration was more pronounced. The vapor phase effect was more pronounced at lower concentrations in most cases. Antibiofilm effect against Salmonella enterica was demonstrated. The relatively strong insecticidal activity was demonstrated with an LC50 value of 21.07% and an LC90 value of 78.21%, making CSEO potentially adequate in the control of agricultural insect pests. Results of cell viability testing showed no effects on the normal MRC-5 cell line, and antiproliferative effects towards MDA-MB-231, HCT-116, JEG-3, and K562 cells, whereas K562 cells were the most sensitive. Based on our results, CSEO could be a suitable alternative against different types of microorganisms as well as suitable for the control of biofilms. Due to its insecticidal properties, it could be used in the control of agricultural insect pests.

Salvia sclarea Essential Oil Chemical Composition and Biological Activities.

Salvia sclarea essential oil (SSEO) has a long tradition in the food, cosmetic, and perfume industries. The present study aimed to analyze the chemical composition of SSEO, its antioxidant activity, antimicrobial activity in vitro and in situ, antibiofilm, and insecticidal activity. Besides that, in this study, we have evaluated the antimicrobial activity of SSEO constituent (E)-caryophyllene and standard antibiotic meropenem. Identification of volatile constituents was performed by using gas chromatography (GC) and gas chromatography/mass spectrometry (GC/MS) techniques. Results obtained indicated that the main constituents of SSEO were linalool acetate (49.1%) and linalool (20.6%), followed by (E)-caryophyllene (5.1%), p-cimene (4.9%), a-terpineol (4.9%), and geranyl acetate (4.4%). Antioxidant activity was determined as low by the means of neutralization of the DDPH radical and ABTS radical cation. The SSEO was able to neutralize the DPPH radical to an extent of 11.76 ± 1.34%, while its ability to decolorize the ABTS radical cation was determined at 29.70 ± 1.45%. Preliminary results of antimicrobial activity were obtained with the disc diffusion method, while further results were obtained by broth microdilution and the vapor phase method. Overall, the results of antimicrobial testing of SSEO, (E)-caryophyllene, and meropenem, were moderate. However, the lowest MIC values, determined in the range of 0.22-0.75 µg/mL for MIC50 and 0.39-0.89 µg/mL for MIC90, were observed for (E)-caryophyllene. The antimicrobial activity of the vapor phase of SSEO (towards microorganisms growing on potato) was significantly stronger than that of the contact application. Biofilm analysis using the MALDI TOF MS Biotyper showed changes in the protein profile of Pseudomonas fluorescens that showed the efficiency of SSEO in inhibiting biofilm formation on stainless-steel and plastic surfaces. The insecticidal potential of SSEO against Oxycarenus lavatera was also demonstrated, and results show that the highest concentration was the most effective, showing insecticidal activity of 66.66%. The results obtained in this study indicate the potential application of SSEO as a biofilm control agent, in the shelf-life extension and storage of potatoes, and as an insecticidal agent.

An In-Depth Study on the Chemical Composition and Biological Effects of Pelargonium graveolens Essential Oil.

The essential oil of Pelargonium graveolens (PGEO) is identified in the literature as a rich source of bioactive compounds with a high level of biological activity. This study aimed to examine the chemical profile of PGEO as well as its antioxidant, antibacterial, antibiofilm, and insecticidal properties. Its chemical composition was analyzed using gas chromatography-mass spectrometry (GC-MS), achieving comprehensive identification of 99.2% of volatile compounds. The predominant identified compounds were ß-citronellol (29.7%) and geraniol (14.6%). PGEO's antioxidant potential was determined by means of DPPH radical and ABTS radical cation neutralization. The results indicate a higher capacity of PGEO to neutralize the ABTS radical cation, with an IC50 value of 0.26 ± 0.02 mg/mL. Two techniques were used to assess antimicrobial activity: minimum inhibitory concentration (MIC) and disk diffusion. Antimicrobial evaluation using the disk diffusion method revealed that Salmonella enterica (14.33 ± 0.58 mm), which forms biofilms, and Priestia megaterium (14.67 ± 0.58 mm) were most susceptible to exposure to PGEO. The MIC assay demonstrated the highest performance of this EO against biofilm-forming S. enterica (MIC 50 0.57 ± 0.006; MIC 90 0.169 ± 0.08 mg/mL). In contrast to contact application, the assessment of the in situ vapor phase antibacterial activity of PGEO revealed significantly more potent effects. An analysis of antibiofilm activity using MALDI-TOF MS demonstrated PGEO's capacity to disrupt the biofilm homeostasis of S. enterica growing on plastic and stainless steel. Additionally, insecticidal evaluations indicated that treatment with PGEO at doses of 100% and 50% resulted in the complete mortality of all Harmonia axyridis individuals.

Determination of Antioxidant, Antimicrobial Activity, Heavy Metals and Elements Content of Seaweed Extracts.

The aim of the research was to determine the antioxidant and antimicrobial activity, determination of chemical elements and heavy metals in seaweed extracts of wakame, arame, dulse, laminaria, kombu, and hijiki. Antioxidant activity was determined by DPPH method and the activity ranged from 0.00 to 2641.34 TEAC. The highest antioxidant activity was observed in kombu (2641.34 TEAC) and arame (2457.5 TEAC). Antimicrobial activity was analyzed by disk diffusion method and MIC method. Three G+ bacteria (Staphylococcus aureus, Enterococcus faecalis, Bacillus subtilis), three G- bacteria (Salmonella enterica, Pseudomonas aeruginosa, Yersinia enterocolitica), and four yeasts (Candida tropicalis, C. krusei, C. glabrata, C. albicans) were used as model organisms. The size of inhibition zones ranged from 0.00 to 8.67 mm. The minimum inhibitory concentrations of the selected seaweeds ranged from MIC50 98.46 (MIC90 100.25) to MIC50 3.43 µL/mL (MIC90 5.26 µL/mL). The content of selected elements was determined in seaweed samples by ICP-OES. The chemical composition of the algae showed differences between species and the presence of heavy metals. Arsenic, cadmium, and aluminum were confirmed. All seaweed samples contained arsenic ranging from 6.6546 to 76.48 mg/kg. Further investigation of seaweeds is needed to identify the active substances present in the algae that are responsible for antioxidant and antimicrobial activity. This study was carried out to evaluate the antimicrobial and antioxidant activity of extracts from five commonly consumed seaweeds for their ability to inhibit selected microorganisms and to determine the health risk due to heavy metals content. Our study contributes to the evidence that seaweeds have antimicrobial and antioxidant activity and seaweed extracts have for pharmacological applications.

Assessment of Chemical Composition and Anti-Penicillium Activity of Vapours of Essential Oils from Abies Alba and Two Melaleuca Species in Food Model Systems.

The possibilities of the practical utilization of essential oils (EOs) from various plant species in the food industry have attracted the attention of the scientific community. Following our previous studies, the antifungal activities of three further commercial EOs, Melaleuca armillaris subsp. armillaris (rosalina; REO), Melaleuca quinquenervia (niaouli; NEO), and Abies alba (fir; FEO), were evaluated in the present research in respect to their chemical profiles, over four different concentrations, 62.5 µL/L, 125 µL/L, 250 µL/L, and 500 µL/L. The findings revealed that the major compounds of REO, NEO, and FEO were linalool (47.5%), 1,8-cineole (40.8%), and α-pinene (25.2%), respectively. In vitro antifungal determinations showed that the inhibition zones of a Penicillium spp. mycelial growth ranged from no inhibitory effectiveness (00.00 ± 00.00 mm) to 16.00 ± 1.00 mm, indicating a very strong antifungal activity which was detected against P. citrinum after the highest REO concentration exposure. Furthermore, the in situ antifungal efficacy of all EOs investigated was shown to be dose-dependent. In this sense, we have found that the highest concentration (500 µL/L) of REO, NEO, and FEO significantly reduced (p < 0.05) the growth of all Penicillium strains inoculated on the bread, carrot, and potato models. These results indicate that the investigated EOs may be promising innovative agents in order to extend the shelf life of different types of food products, such as bread, carrot and potato.

Microbiological and Physicochemical Composition of Various Types of Homemade Kombucha Beverages Using Alternative Kinds of Sugars.

Kombucha is a beverage made by fermenting sweetened tea with a symbiotic culture of yeast and bacteria. Literature data indicate that the kombucha beverage shows many health-promoting properties such as detoxification, chemo-preventive, antioxidant, antimicrobial, antifungal, and general strengthening. The research conducted focuses on the analysis of polyphenolic compounds formed in the fermentation process using ultra-efficient liquid chromatography, as well as on checking the antimicrobial properties of kombucha against pathogenic bacteria and yeasts found in food. Analysis of the composition of the tea mushroom (SCOBY) microflora using the MALDI TOF MS Biotyper mass spectrometer showed 8 species of bacteria and 7 species of yeasts. In vitro studies confirm the bactericidal and bacteriostatic properties of fermented kombucha beverages, with white and green tea beverages showing the highest antibacterial activity. The bacteria Staphylococcus aureus and yeast Candida albicans were the most sensitive to the effects of kombucha tea beverages. UPLC chromatographic analysis confirmed the presence of 17 bioactive compounds in kombucha beverages that can affect human health. The analyses conducted were aimed at indicating the best recipe and conditions to prepare a kombucha beverage, which allowed the selection of the version with the best health-promoting properties. Fermented kombucha teas contain many elements such as aluminium, calcium, iron, potassium, magnesium, sodium, phosphorus, and sulphur.

Assessment of Ocimum basilicum Essential Oil Anti-Insect Activity and Antimicrobial Protection in Fruit and Vegetable Quality.

Basil (Ocimum basilicum) is a commonly used herb; it also contains essential oils and other valuable compounds. The basil oil obtained has a pleasant aroma, but also a broad spectrum of biological activity. This work reports on the chemical composition, antioxidant, antimicrobial and anti-insect activity in vitro and in situ of Ocimum basilicum essential oil (OBEO) obtained by steam distillation of fresh flowering plants. Gas chromatography-mass spectrometry, DPPH, agar and disc diffusion and vapor phase methods were used to analyze the OBEO properties. The analysis of the chemical composition of OBEO showed that its main components were methyl chavicol (88.6%), 1,8-cineole (4.2%) and α-trans-bergamotene (1.7%). A strong antioxidant effect was demonstrated at the level of 77.3%. The analysis of antimicrobial properties showed that OBEO exerts variable strength of inhibiting activity against various groups of microorganisms. The growth inhibition zones ranged from 9.67 to 15.33 mm in Gram-positive (G+) and Gram-negative (G-) bacteria and from 5.33 to 7.33 mm in yeast. The lowest measured minimal inhibition concentration (MIC) was 3.21 µL/mL against Gram-negative Azotobacter chrococcum and Gram-positive Micrococcus luteus. The antimicrobial activity of in situ vapor phase of OBEO was also confirmed on apples, pears, potatoes and kohlrabi. The highest insecticidal activity against Pyrrhocorisapterus, observed at the concentration of 100%, caused the death of 80% of individuals. Due to its broad spectrum of activity, OBEO seems an ideal candidate for preserving fruit and vegetables.

Technological and Sensory Quality and Microbiological Safety of RIR Chicken Breast Meat Marinated with Fermented Milk Products.

The aim of the study was to determine the effect of marinating with fermented milk products (buttermilk and sour milk) on the physical characteristics, microbiological quality, and sensory acceptability of Rhode Island Red (RIR) hen meat after the first year of laying use. The hen breast meat was marinated with fermented dairy products, buttermilk and sour milk, by the immersion method for 12 h at 4 °C. The assessed features included the quality of raw and roasted marinated and non-marinated meat in terms of physical characteristics (marinade absorption, water absorption, pH, L*, a*, b* colour, shear strength, texture profile analysis (TPA) test), microbiological parameters, and sensory characteristics. Bacteria were identified by the mass spectrometry method (MALDI-TOF MS Biotyper). Marinating meat with fermented dairy products lightened the colour, decreased the value of shear force, reduced hardness and chewiness, and limited the growth of aerobic bacteria and Pseudomonas spp. Additionally, after heat treatment, the number of identified aerobic bacteria families in the marinated in buttermilk and marinated in sour milk groups was smaller than in the non-marinated muscle group. The sensory evaluation showed a beneficial effect of marinating with buttermilk and sour milk on the tenderness, juiciness, and colour of roasted meat.

Combined Effect of Vacuum Packaging, Fennel and Savory Essential Oil Treatment on the Quality of Chicken Thighs.

The aim of the present work was to evaluate the microbiological quality of chicken thighs after treatment by fennel (Foeniculum vulgare) and savory (Satureja hortensis) essential oil, stored under vacuum packaging (VP) at 4 ± 0.5 °C for a period of 16 days. The following treatments of chicken thighs were used: Air-packaging control samples (APCS), vacuum-packaging control samples (VPC), vacuum-packaging (VP) control samples with rapeseed oil (VPRO), VP (vacuum-packaging) with fennel essential oil at concentrations 0.2% v/w (VP + F), and VP with savory essential oil at concentration 0.2% v/w (VP + S). The quality assessment of APCS, VPC, VPRO, VP + F and VP + S products was established by microbiological analysis. The microbiological parameters as the total viable counts of bacteria of the Enterobacteriaceae family, lactic acid bacteria (LAB), and Pseudomonas spp. were detected. Bacterial species were identified with the MALDI-TOF MS Biotyper. The combination of essential oils and vacuum packaging had a significant effect (p < 0.05) on the reduction of total viable counts (TVC) compared with control group without vacuum packaging and the untreated control group. Though 15 genera and 46 species were isolated with scores higher than 2.3 from the chicken samples.

Effect of addition of wild garlic (Allium ursinum) on the quality of kefirs from sheep's milk.

BACKGROUND: Sheep’s milk has a high content of total solids, which qualifies it as a very good raw material for the production of fermented milk drinks. Currently, there are commercially produced kefirs and yogurts from sheep’s milk in the countries of the Mediterranean region. The growing interest in the consumption of these products is justified not only by their taste merits, but also because of their health-promoting proper- ties. The aim of the present study was to determine the effect of the addition of 1% of lyophilized wild garlic powder on the properties of kefirs from sheep’s milk. METHODS: Sheep’s milk was pasteurized (85°C, 30 min), cooled down, enriched with 1% of freeze-dried wild garlic powder, inoculated with a Commercial VITAL kefir culture and fermented for 16 hours (26°C). The influence of wild garlic on acidity (pH, °SH), syneresis (%), texture (TPA test), colour (L*a*b*) and the sensory profile of kefirs was conducted. RESULTS: Wild garlic could be used as a taste and flavour modifier in the production of kefir from sheep’s milk. The addition of 1% of freeze-dried wild garlic slowed down the fermentation of kefir, changed colour and reduced syneresis. CONCLUSIONS: Wild garlic could be used as a valuable supplement and a modifier of taste and flavour in kefir from sheep’s milk.

The antioxidant and antimicrobial activity of essential oils against Pseudomonas spp. isolated from fish.

Natural products of plant origin, which include essential oils (EO) could be used as a growth inhibitor of pathogenic and spoilage microflora in food. The objective of this study was to determine the antibacterial and antioxidant activity of 21 EO against 10 Pseudomonas species isolated from freshwater fish. The chemical composition of EO was determined by gas chromatography/mass spectrometry. The disc diffusion method and detection of minimum inhibitory concentration (MIC) were used for the determination of the antimicrobial activity. All the EO tested exhibited antimicrobial activity, however, Cinnamomum zeylanicum EO was the most effective against Pseudomonas spp. both according to the disc diffusion and MIC methods. The EOs of Cymbopogon nardus, Origanum vulgare, Foeniculum vulgare and Thymus serpyllum showed the highest antioxidant activity of 93.86 µg, 83.47 µg, 76.74 µg and 74.28 µg TEAC/mL. Application of EO could be an effective tool for inhibition of growth of Pseudomonas spp. on fish.

Identification of lactic acid bacteria isolated from wine using real-time PCR.

Different lactic acid bacteria strains have been shown to cause wine spoilage, including the generation of substances undesirable for the health of wine consumers. The aim of this study was to investigate the occurrence of selected species of heterofermentative lactobacilli, specifically Lactobacillus brevis, Lactobacillus hilgardii, and Lactobacillus plantarum in six different Slovak red wines following the fermentation process. In order to identify the dominant Lactobacillus strain using quantitative (real time) polymerized chain reaction (qPCR) method, pure lyophilized bacterial cultures from the Czech Collection of Microorganisms were used. Six different red wine samples following malolactic fermentation were obtained from selected wineries. After collection, the samples were subjected to a classic plate dilution method for enumeration of lactobacilli cells. Real-time PCR was performed after DNA extraction from pure bacterial strains and wine samples. We used SYBR® Green master mix reagents for measuring the fluorescence in qPCR. The number of lactobacilli ranged from 3.60 to 5.02 log CFU mL(-1). Specific lactobacilli strains were confirmed by qPCR in all wine samples. The number of lactobacilli ranged from 10(3) to 10(6) CFU mL(-1). A melting curve with different melting temperatures (T(m)) of DNA amplicons was obtained after PCR for the comparison of T(m) of control and experimental portions, revealing that the most common species in wine samples was Lactobacillus plantarum with a T(m) of 84.64°C.

Construction and fed-batch cultivation of Candida famata with enhanced riboflavin production.

Riboflavin (vitamin B2) is an essential nutrition component serving as a precursor of coenzymes FMN and FAD that are involved mostly in reactions of oxidative metabolism. Riboflavin is produced in commercial scale and is used in feed and food industries, and in medicine. The yeast Candida famata (Candida flareri) belongs to the group of so called "flavinogenic yeasts" which overproduce riboflavin under iron limitation. Three genes SEF1, RIB1 and RIB7 coding for a putative transcription factor, GTP cyclohydrolase II and riboflavin synthase, respectively were simultaneously overexpressed in the background of a non-reverting riboflavin producing mutant AF-4, obtained earlier in our laboratory using methods of classical selection (Dmytruk et al. (2011), Metabolic Engineering 13, 82-88). Cultivation conditions of the constructed strain were optimized for shake-flasks and bioreactor cultivations. The constructed strain accumulated up to 16.4g/L of riboflavin in optimized medium in a 7L laboratory bioreactor during fed-batch fermentation.

The StepOne real-time polymerase chain reaction detection of Salmonella sp., Salmonella enterica ser. typhimurium and enteritidis in milk and meat.

The aim of this study was to follow contamination of ready to eat milk and meat products with Salmonella spp. by using the StepOne real-time polymerase chain reaction (PCR). Classical microbiological methods for detection of foodborne bacteria involve the use of pre-enrichment and/or specific enrichment, following isolation of bacteria in solid media and the final confirmation by biochemical and/or serological tests. We used the PrepSEQ Rapid Spin Sample Preparation Kit for isolation of DNA and MicroSEQ® Salmonella spp. Detection Kit for pursuance of the real-time PCR (Applied Biosystems). In samples without incubation we detected strain of Salmonella sp. in 5 out of 25 samples (swabs), as well as in the internal positive control (IPC), which was positive in all samples. This StepOne real-time PCR assay is extremely useful for any laboratory equipped by real-time PCR. It is a fast, reproducible, simple, specific and sensitive way to detect nucleic acids, which could be used in clinical diagnostic tests in the future. Our results indicated that real-time PCR assay developed in this study could sensitively detect Salmonella spp. in ready-to-eat food. This could prevent infection caused by Salmonella, and also could benefit food manufacturing companies by extending their product's shelf-life as well as saving the cost of warehousing their food products while awaiting pathogen testing results.