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The aim of this study was to isolate and identify yeast species in wine samples from five different countries, and determine the genera of spoilage yeasts. One hundred and thirty four (no. 134) wine samples (86 white and 48 red wine); (30 bottled, 104 tank wine) were used in this study from five countries Slovakia (no. 53), Czech Republic (no. 17), Hungary (no. 6), Spain (no. 32) and Italy (no. 26). Yeasts were cultivated on Malt extract agar supplemented with bromocresol green. After cultivation, yeasts were identified on species level by MALDI-TOF mass spectrometry. From total 285 isolates, we identify 10 genera of yeasts, Saccharomyces (97 isolates, 1 species), Pichia (120 isolates, 5 species), Kregervanrija (6 isolates, 1 species), Zygosaccharomyces (32 isolates, 2 species), Candida (13 isolates, 2 species), Rhodotorula (4 isolates, 1 species), Wickerhamomyces (7 isolates, 1 species) Meyerozyma (4 isolates, 1 species) and genera with only 1 isolale and species: Torulaspora, and Cyberlindnera.

The aim of this study was to investigate the antioxidant and antimicrobial effect of two selected plants: Tussilago farfara L. and Cetraria islandica L. The antioxidant activity was evaluated by DPPH and phosphomolybdenum method, total polyphenol content with Folin – Ciocalteu reagent, flavonoids content by aluminium chloride method. The detection of antimicrobial activity was carried out by disc diffusion method and evaluation of minimal inhibition concentration against five species of Gram-negative bacteria: Escherichia coli CCM 3988, Klebsiella pneumoniae CCM 2318, Salmonella enterica subsp. enterica CCM 3807, Shigella sonei CCM 1373, Yersinia enterocolitica CCM 5671 and five Gram-positive bacteria: Bacillus thuringiensis CCM 19, Clostridium perfringens CCM 4435, Haemophilus influenzae CCM4456, Listeria monocytogenes CCM 4699, Stapylococcus aureus subsp. aureus CCM 2461. Tussilago farfara showed stronger antioxidant activity (DPPH method – 7.67 mg TEAC – Trolox equivalent antioxidant capacity per g of sample; phosphomolybdenum method – 111.67 mg TEAC per g of sample), total polyphenol (11.92 mg GAE – galic acid equivalent per g of sample) and flavonid content (47.62 μg QE – quercetin equivalent per g of sample) with compare to Centraria islandica. Tussilago farfara also showed the higher antimicrobial activity against Gram-negative bacteria confirmed by both methods – disc diffusion and minimal inhibitory concentration.

Quantitative microbiological analysis by dilution plating method of pollen and additional male and female catkins, leaves of Betula verrucosa Ehrh. and its two cultivars: ‘Purpurea’ and ‘Youngii’ relatively of flowering period of Betula has been realized with the aim to provide new knowledge of the microbiological quality of anemophilous pollen for processing and its further application. Qualitative microbiological analysis with MALDI-TOF MS Biotyper was used in the identification of aerobic, anaerobic mesophilic bacteria and coliforms. Mixed microbiota was determined, consisting of aerobic (4.68–4.89 log cfu/g) and anaerobic (3.30–3.48 log cfu/g) mesophilic bacteria, lactobacilli (0–3.48 log cfu/g), coliform bacteria (0–4.57 log cfu/g), fungi and yeast (3.78–3.95 log cfu/g) on the pollen grains, that indicates acceptable quality in comparison with the microbiological quality parameters for bee pollen. Pantoea agglomerans was found associated with pollen of Betula verrucosa Ehrh. Recommendations on the collection of anemophilous pollen were established.

The aim of this study was to determine biological activity of plants with adaptogenic effect: Panax ginseng Mayer., Withania somnifera L., Eleuterococcus senticosus Rupr. et Maxim., Astragallus membranaceus Fisch. and Codonopsis pilosulae Franch. The antioxidant activity was detected by DPPH and phosphomolybdenum method, total polyphenol content with Folin – Ciocalteu reagent, flavonoids content by aluminium chloride method. The detection of antimicrobial activity was carried out by disc diffusion method against three species of Gram-negative bacteria: Escherichia coli CCM 3988, Salmonella enterica subsp. enterica CCM 3807, Yersinia enterocolitica CCM 5671 and two Gram-positive bacteria: Bacillus thuringiensis CCM 19, Stapylococcus aureus subsp. aureus CCM 2461. Results showed that plants with adaptogenic effect are rich for biologically active substances. The highest antioxidant activity by DPPH method was determined in the sample of Eleuterococcus senticosus (3.15 mg TEAC – Trolox equivalent antioxidant capacity per g of sample) and by phosphomolybdenum method in the sample of Codonopsis pilosulae (188.79 mg TEAC per g of sample). In the sample of Panax ginseng was measured the highest content of total polyphenols (8.10 mg GAE – galic acid equivalent per g of sample) and flavonoids (3.41 μg QE – quercetin equivalent per g of sample). All samples also showed strong antimicrobial activity with the best results in Panax ginseng and Withania somnifera in particular for species Yersinia enterocolitica CCM 5671 and Salmonella enterica subsp. enterica CCM 3807. The analyzed species of plant with high value of biological activity can be used more in the future, not only in food, but also in cosmetics and pharmaceutical industries.

Antibiotic resistance of bacteria is very big problem in clinical and veterinary medicine in recent year. This problem is increasing by the exposure of bacteria against antibiotics. The most spread of resistance is resistance against penicillins antibiotics. Ampicillin is common use antibiotic in the both medicine. Therefore, detection of antibiotics and antibiotic resistance are very important for prevention of spread of antibiotics in the environment and resistant bacteria too. Therefore the aim of this study was detection of ampicillin, its sodium salts and alkaline hydrolysis products by MALDI-TOF Mass Spectrometry. For detection of pure ampicillin, its sodium salts and alkaline hydrolysed products MALDI-TOF MS (Matrix Assisted Light Desorption Ionization – Time of Flight Mass Spectrometry) Microflex LT in positive ion mode was used. Determined mass spectra were observed by FlexAnalysis software. The mass spectra and molecular weight of ampicillin, its sodium salts and hydrolysed products were compared with theoretical molecular weight of these compounds. The results showed that its possible to detect ampicillin, its sodium salts and hydrolyzed products by MALDI-TOF MS in positive ion mode with some correction in analytical software. This knowledge is resulting that MALDI-TOF MS detection method can be useful for detection of ampicillin from different kind of environment samples and its possible to detect ampicillin resistance mechanism (enzymatic destruction) indirectly, because resistance against penicillins is enzymatic hydrolysis of beta-lactam core. Also the most important issue is that method is very quickly and cheap.

Medicinal plants are traditionally used for the treatment in human medicine. The antibacterial activity of ethanol extract of four plant species (Achillea millefolium L., Agrimonia eupatoria, Melissa officinalis and Tilia platyphyllos) applied in the traditional medicine in Slovakia were tested. Extracts of certain parts of these plants were tested in vitro against six bacterial species (Bacillus cereus, Enterococcus faecalis, Escherichia coli, Lactobacillus brevis, Lactobacillus hilgardii and Serratia rubidea) strains using the disc diffusion method and microbroth dilution method. Minimum inhibitory concentrations for the extracts against all the microorganisms were determined by serial dilutions. All the extracts demonstrated antibacterial activity against Gram-positive bacteria and partially against Gram-negative bacteria.

The aim of the present work was monitoring chicken thighs microbiological quality after treatment by ethylenediaminetetraacetate (EDTA), coriander (Coriandri aetheroleum) and spearmint (Menthae spicata aetheroleum) essential oil, stored under vacuum packaging, at 4±0.5°C for a period of 16 days. The following treatments of chicken thighs were used: Air-packaging control samples, control vacuum-packaging samples, vacuum-packaging with EDTA solution 1.50% w/w, control samples, vacuum-packaging with Menthae crispae aetheroleum essential oil at concentrations 0.2% v/w and vacuum-packaging with Coriandri aetheroleum essential oil at concentration 0.2% v/w. The quality assessment of all samples was established by microbiological analysis. The microbiological parameters as the total viable count, Enterobacteraceae genera counts, lactic acid bacteria and Pseudomonas spp. were detected. The results of this present study suggest the possibility of application the essential oil of Coriandri aetheroleum and Menthae spicata aetheroleum as natural food preservatives and potential sources of antimicrobial ingredients for food industry.

The aim of this study was to examine the effect of different feed additives on the Enterobacteriaceae genera colonization of chicken (Coob 500) gastrointestinal tract. The Agolin Poultry, AgolinTannin Plus, Biostrong 510+FortiBac and Agolin acid were administered to forth feed mixtures and Biocitro to drinking water in various amounts except of the control group. The addition of 1.5 g Agolin Poultry to 15 kg of feed was included in the first experimental group, the addition of 7.5 g Agolin Tannin Plus to 15 kg of feed in the second experimental group, the addition of 15g Biostrong510+FortiBac to 15 kg of feed the third experimental group, 15 g in the fourth experimental group of Agolin acid to 15 kg of feed and 10 ml fifth experimental group of drinking water. The highest count of Enterobacteriaceae genera counts was found in the control group. The lower count of Enterobacteriaceae genera counts was found in the second experimental group where 7.5 g of Agolin Tannin Plus to 15 kg was added to feed mixture.

The present study was undertaken to provide detailed information about the effect of different feed additives on the enterococci colonization of chicken gastrointestinal tract. The Agolin Poultry, Agolin Tannin Plus, Biostrong 510+FortiBac and Agolin acid were administered to forth feed mixtures and Biocitro to drinking water in various amounts except of the control group. The addition of 1.5 g Agolin Poultry to 15 kg of feed was included in the first experimental group, the addition of 7.5 g Agolin Tannin Plus to 15 kg of feed in the second experimental group, the addition of 15g Biostrong510+FortiBac to 15 kg of feed the third experimental group, 15 g in the fourth experimental group of Agolin acid to 15 kg of feed and 10 ml fifth experimental group of drinking water. The lowest count of enterococci counts was found in the control group. The highest count of enterococci counts was found in the second experimental group where 7.5 g of Agolin Tannin Plus to 15 kg was added to feed mixture. For the identification of eterococci species the molecular method was used.

Clostridium is an anaerobic, endospore forming Gram-positive bacillus genus containing many important pathogenic species. Many naturally occurring compounds present in plants, herbs, and spices have been shown to possess antimicrobial effect against foodborne pathogens. In the present study, the antimicrobial activities of the bee collected pollen samples were investigated. The antimicrobial activities were determined by using agar disc diffusion method against Clostridium genus. Antibacterial activity was assessed on the clostridia: Clostridium butyricum, Clostridium hystoliticum, Clostridium intestinale, Clostridium perfringens and Clostridium ramosum. The results of the disk diffusion method showed very different activity against all tested strains of clostridia. The best antimicrobial activity of bee collected pollen against C. butyricum and C. perfringens were found.

Droseracae spp. is widely used in folk medicine. In the present study, the antimicrobial activities of the four Drosera rotundifolia L. (D8.11, D15.12, 18.10, 8.11) samples were investigated. The antimicrobial activities were determined by using agar disc diffusion method against grampositive bacteria (Bacillus thurigiensis, Staphylococcus aureus, Listeria monocytogenes) and gramnegative bacteria (Yersinia enterocolitica, Salmonella enteritidis). The results of the disk diffusion method showed very different activity against all tested strains of microorganisms. The best antimicrobial activity of ethanolic extract Drosera rotundifolia L. against Salmonella enteritidis was found at Drosera rotundifolia (D8.11).

The aim of this study was to screen 15 essential oils of selected plant species, viz. [i]Lavandula angustifolia[/i], [i]Carum carvi[/i], [i]Pinus mungo var. pulmilio[/i], [i]Mentha piperita[/i], [i]Chamomilla recutita[/i] L.,[i] Pinus sylvestris[/i], [i]Satureia hortensis[/i] L., [i]Origanum vulgare[/i] L., [i]Pimpinella anisum[/i], [i]Rosmarinus officinali[/i]s L., [i]Salvia officinalis[/i] L., [i]Abietis albia etheroleum[/i], [i]Chamomilla recutita[/i] L. [i]Rausch[/i], [i]Thymus vulgaris[/i] L., [i]Origanum vulgare[/i] L. for antifungal activity against five [i]Penicillium[/i] species: [i]Penicillium brevicompactum[/i], [i]Penicillium citrinum[/i], [i]Penicillium crustosum[/i], [i]Penicillium expansum[/i] and [i]Penicillium griseofulvum[/i]. The method used for screening included the disc diffusion method. The study points out the wide spectrum of antifungal activity of essential oils against [i]Penicillium[/i] fungi. There were five essential oils of the 15 mentioned above which showed a hopeful antifungal activity: [i]Pimpinella anisum[/i], [i]Chamomilla recutita[/i] L., [i]Thymus vulgaris[/i], [i]Origanum vulgare[/i] L. The most hopeful antifungal activity and killing effect against all tested penicillia was found to be [i]Origanum vulgare[/i] L. and [i]Pimpinella anisum[/i]. The lowest level of antifungal activity was demonstrated by the oils [i]Pinus mungo var. pulmilio[/i], [i]Salvia officinalis[/i] L., [i]Abietis albia etheroleum[/i], [i]Chamomilla recutita[/i] L.[i] Rausch[/i], [i]Rosmarinus officinalis[/i].

The effect of Salvia officinalis L. essential oil as well as vacuum packaging in extending the shelf life of fresh chicken’s breast meat stored at 4 °C was investigated. In a preliminary experiment Salvia officinalis L. essential oil were used at concentrations 2% v/w while vacuum packaging. Microbiological properties of fresh chicken breast meat were monitored over a 16 days period. For this experiment three groups were used. First group was control with air packaging second was with vacuum packaging condition and was treated with essential oil on the surface of fresh chicken breast meat. From the microbiological indicators in this experiment total count of bacteria and coliform bacteria were observed. The total count of bacteria on the meat after killing animals was 2.97 log cfu.g-1 and number of coliform bacteria was 0.33 log cfu.g-1. The total count of bacteria on the chicken breast meat after 4, 8, 12 and 16 days gradually increased. The same number of coliform bacteria in extending self-life gradually increased. The highest number of both groups of microorganisms was in the control group with air condition and lowest number of both bacterial groups was in the group with salvia essential oil treatment.

The aim of this study was to follow the contamination of food with Listeria monocytogenes by using Step One real time polymerase chain reaction (RT PCR). We used the PrepSEQ Rapid Spin Sample Preparation Kit for isolation of DNA and SensiFAST SYBR Hi-ROX Kit for the real-time PCR performance. In 20 samples of food of animal origin with incubation were detected strains of Listeria monocytogenes in 9 samples (swabs). Eleven samples were negative. Our results indicated that the real-time PCR assay developed in this study could sensitively detect Listeria monocytogenes in food of animal origin without incubation. This could prevent infection caused by Listeria monocytogenes, 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. The rapid real-time PCR-based method performed very well compared to the conventional method. It is a fast, simple, specific and sensitive way to detect nucleic acids, which could be used in clinical diagnostic tests in the future.

The general objective of this study was to examine the effect of bee product on the Enterococci colonization of chickens. Bee products were administered to both feed mixtures in various amounts in addition to the control group. First experimental group was with propolis in feed mixture with the addition of 200 mg propolis per 1 kg of compound and second group was with pollen with the addition of 250 mg pollen per 1 kg of compound. In this experiment, quantitative counts of Enterococci in ceca of 49-day-old chicken (Ross 308) using classical and MALDI TOF MS Biotyper method were investigated. Counts of Enterococci on Slanetz-Bartley agar were monitored. Enterococcus cells, isolated from gastrointestinal tract, were detected using MALDI TOF MS Biotyper. Counts of CFU of Enterococci were compared in experimental and control treatments, respectively. The lowest count was detected in the control experimental group. The highest count was detected in the first experimental group where was 200 mg of propolis added to 1 kg of feed mixture. Using MALDI TOF MS Biotyper, we identified the species range of the genera Enterococcus in the intestinal tract of broiler. Detected species from the genus Enterococcus were: E. avium, E. casseliflavus, E cecorum, E. faecalis, E. faecium, E. gallinarum, E. hirae and E. malodoratus. In the experimental groups (caecal samples) were most frequent species of E. avium E. faecium and E. gallinarum.

The objective of the present research was to develop vacuum packaging incorporated with Rosmarinus officinalis L. essential oil treatment of fresh chicken breast meat. For this purpose, fresh chicken’s meat breast samples were divided into three groups. First group was kept as a control group with air packaging, others one was with vacuum packaging of samples and last one group was treated with rosemary essential oil. All fresh chickens’ breast meat samples were stored at 4°C and microbiological evaluation was conducted at intervals of 0, 4, 8, 12 and 16 days post-storage for lactic acid bacteria and Pseudomonas aeroginosa counts. Rosmarinus officinalis L. essential oil 2%, significantly reduced lactic acid bacteria and Pseudomonas aeroginosa counts in the fresh chickens breast meat samples. The results indicated that 2% essential oil improved the microbiological quality and prolonged the shelf-life of the fresh chicken’s breast meat to sixteen days of retail displayed at 4°C. The results obtained in this study point to the necessity of continuing investigations to determine the dose of rosemary preparations that would inhibit the growth of microflora being the most frequent cause of raw materials and products spoilage.

The general objective of this study was to examine the effect of bee products on the lactobacilli colonization of chickens. Bee products were administered to both feed mixtures in various amounts in addition to the control group. First experimental group was with propolis in feed mixture with the addition of 400 mg propolis per 1 kg of compound and second group was with pollen in feed mixture with the addition of 450 mg pollen per 1 kg of compound. In this experiment, quantitative counts of lactobacilli in ceca of 49-day-old chicken (Ross 308) using classical and FISH method were investigated. Counts of lactobacilli on MRS agar were monitored. To check the reliability of traditional methods of cultivation samples were evaluated by fluorescence in situ hybridization (FISH). Lactobacillus cells, isolated from gastrointestinal tract, were detected after hybridization of fluorescently labeled probe with bacterial cells. Counts of CFU of lactobacilli were compared in experimental and control treatments, respectively. The lowest count was detected in the control experimental group. The highest count was detected in the third experimental group where was 450 mg of pollen added to 1 kg of feed mixture. Using Real-time PCR method, we identified the species range of the genera Lactobacillus in the intestinal tract of broiler. Detected species from the genus Lactobacillus were L. crispatus, L. salivarius and L. acidophilus.

The aim of this study was follow the contamination of ready to eat milk and meat products with Salmonella spp. by using the Step One real-time PCR. Classical microbiological methods for detection of food-borne bacteria involve the use of pre-enrichment and/or specific enrichment, followed by the isolation of the bacteria in solid media and a final confirmation by biochemical and/or serological tests. We used the PrepSEQ Rapid Spin Sample Preparation Kit for isolation of DNA and SensiFAST SYBR Hi-ROX Kit for the real-time PCR performance. In the investigated samples without incubation we could detect strain of Salmonella sp. in five out of twenty three samples (swabs). This Step One real-time PCR assay is extremely useful for any laboratory in possession of a 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 the Step One real-time PCR assay developed in this study could sensitively detect Salmonella spp. in ready to eat food.