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The chemical composition as well as the antioxidant and antimicrobial activities of two EtOH extracts of propolis (PEEs) from Slovenia were determined. EtOH was used as extracting solvent at 70 and 96%, providing the extracts PEE70 and PEE96, respectively. The extraction with 70% EtOH was more efficient than that with 96% EtOH, as the PEE70 was richer in total phenolic compounds than the PEE96. The Slovenian propolis was characterized by different phenolic acids and flavonoids. The PEE96 was slightly richer in three specific compounds, i.e., caffeic acid, ferulic acid, and luteolin, while all other substances detected showed higher contents in the PEE70. The PEE70 showed a stronger reducing power and ability to scavenge free radicals and metal ions than the PEE96. Both PEEs were in the main more effective against Gram-positive bacteria than against fungi and Gram-negative bacteria like Salmonella and Escherichia coli, with the exception of Campylobacter. The PEE96 decreased the intracellular oxidation in Saccharomyces cerevisiae in a dose-dependent manner. The antimicrobial activities and antioxidant properties were related to the total phenolic contents. The two PEEs have the potential for use as natural antimicrobial and antioxidant additives in foods.

Repetitive element sequence-based PCR (rep-PCR) was used to generate DNA fingerprints for Listeria spp. Two primer sets (REP 1R-I REP 2-I and ERIC 1R ERIC 2) used in respectively REP- and ERIC-PCR revealed that bacteria of the genus Listeria possess short repetitive extragenic palindromic elements and enterobacterial repetitive intergenic consensus sequences. Specific band profiles obtained by ERIC-PCR enabled the identification of Listeria species. With both REP- and ERIC-PCR the L. monocytogenes serotypes 1/2a, 1/2b, 1/2c, 3b and 4b could be clearly distinguished from each other. Within the serotype 1/2a, REP-PCR showed a higher discriminative potential than ERIC-PCR and a comparable discriminative potential as RAPD combining 3-4 primers.

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Listeria monocytogenes strains possess short repetitive extragenic palindromic (REP) elements and enterobacterial repetitive intergenic consensus (ERIC) sequences. We used repetitive element sequence-based PCR (rep-PCR) to evaluate the potential of REP and ERIC elements for typing L. monocytogenes strains isolated from humans, animals, and foods. On the basis of rep-PCR fingerprints, L. monocytogenes strains were divided into four major clusters matching origin of isolation. rep-PCR fingerprints of human and animal isolates were different from those of food isolates. Computer evaluation of rep-PCR fingerprints allowed discrimination among the tested serotypes 1/2a, 1/2b, 1/2c, 3b, and 4b within each major cluster. The index of discrimination calculated for 52 epidemiologically unrelated isolates of L. monocytogeneswas 0.98 for REP- and ERIC-PCR. Our results suggest that rep-PCR can provide an alternative method for L. monocytogenes typing.

Reduction or elimination of chemically synthesized additives from foods is a current demand in food industry. A new approach to prevent the proliferation of microorganisms or protect food from oxidation is the use of essential oils or plant extracts as natural additives in foods. We have studied antimicrobial activity of rosemary extracts (Rosmarinus officinalis L.) against different species of Listeria and against different strains of L. monocytogenes. We used two extracts of rosemary, VivOX 20 and VivOX 40 (Vitiva d.d., Slovenia) containing different levels of carnosic acid. We wanted to proof an antimicrobial activity of selected rosemary extracts with two most commonly used methods: disc diffusion method and broth dilution method. With the disc diffusion method we have obtained the inhibition zone and at the lowest concentrations, where no visible bacterial growth was recorded, were assumed as minimal inhibitory concentration values (MIC). We determined MIC values in the ranges from 625 μg extract/ml EtOH to 5000 μg extract/ml EtOH for VivOX 20 and from 312.5 μg extract/ml EtOH do 2500 μg extract/ml EtOH for VivOX 40 in the medium. We have established that the resistance of Listeria species against rosemary extracts depends on: selected extract, selected concentration, various species and strain of Listeria. With broth dilution method we have determined minimal bactericidal concentration (MBC), as the concentration giving 0.1 % bacterial survival. With this method we have tested two strains of L. monocytogenes and in determinate MBC values in the range from 15.63 µg/ml TSB to 98.5 µg/ml TSB for both tested extracts. Results have confirmed our assumption that resistance of Listeria against rosemary extracts depended on the selected strain.

Aims:  We describe a real‐time quantitative multiplex polymerase chain reaction (qmPCR) assay to identify and discriminate between isolates of Campylobacter jejuni and Campylobacter coli. Methods and Results:  Two novel sets of primers and hydrolysis probes were designed to amplify the unique DNA sequences within the hipO, ccoN and cadF genes that are specific to Camp. jejuni and Camp. coli. Using the designed optimized qmPCR assay conditions, the amplification efficiency is in range from 108 to 116%. These qmPCR assays are highly specific for Camp. jejuni and Camp. coli, as seen through testing of 40 Campylobacter strains and 17 non‐Campylobacter strains. In chicken juice and tap water models spiked with known quantities of Camp. jejuni, qmPCR detected 102–103 CFU ml−1 within 4 h. Conclusions:  The qmPCR assays developed in this study provide reliable and simultaneous detection and quantification of Camp. jejuni and Camp. coli, with good amplification reaction parameters. Significance and Impact of the Study:  Following further validation, the qmPCR assay reported here has the potential to be applied to various sample types as an alternative and rapid methodology.

The aim of this study was the assessment of bioaerosols in different indoor environments. The study was performed using volumetric culture plate air sampling to determine the concentration of viable bioaerosols. The results revealed that 60.9% of air samples from public places and all air samples collected in food processing plants had unacceptable levels of micro-organisms. This was based on a suggested standard which proposes that concentrations of viable micro-organisms should be no higher than 300 CFU · m-3. More detailed study of the bioaerosols collected showed that the main parameters of interest for indoor air quality were fungi and further characterization allowed identification of the genera present in a particular place. Acceptable indoor air quality, by the above standard, was found at a university faculty, in a fast food restaurant, a cultural centre, a health centre and a hospital, while it was not acceptable in meat-, mustard-, olive- and infant food-processing plants because the concentrations of moulds were higher than 150 CFU· m -3 and the predominant genus identified was Penicillium.