Natural fruits, flowers, honey, and honeybees harbor Helicobacter pylori-positive yeasts.

BACKGROUND: For controlling Helicobacter pylori infection in humans, its environmental reservoir should be determined. In this study, yeast isolates from an isolated village in Iran were studied for the intracellular occurrence of H. pylori. MATERIALS AND METHODS: In this study, yeasts were isolated from 29 samples, including oral swabs from villagers (n = 7), flowers and fruits (n = 6), honey and honeybees (n = 12) and miscellaneous samples (4). Yeasts were classified into 12 RFLP groups and identified by amplification of 26S rDNA and sequencing. DNA extracted from the yeast cells was examined for the presence of H. pylori using PCR. RESULTS: Of the 29 yeasts, 27 were members of different genera of Ascomycete. H. pylori was detected in 5 of 9 Candida (55.5%), 4 of 5 Komagataella (80%), 3 of 4 Pichia (100%), 2 of 2 Cytobasidia (100%), 2 of 2 Hansenia (100%), 1 of 1 Meyerozyma (100%) and 2 of 3 not sequenced (66.6%) yeasts. Distribution of 19 of 29 (65.5%) H. pylori-positive yeasts within 4 groups was as follows: 1 of 7(14.3%) in oral swabs, 5 of 6 (83.3%) in flowers and fruits, 10 of 12 (83.3%) in honey and the bee group and 3 of 4 (75%) in miscellaneous. CONCLUSIONS: Different genera of osmotolerant yeasts from flowers, fruits, honey, and honeybees contained H. pylori in their vacuole. High frequency of H. pylori-positive yeasts in these samples might be related to their high sugar content. Insects such as honeybees that facilitate transfer and easy access of these yeasts to nectars serve as the main reservoirs of these yeasts, playing an important role in their protection and dispersal. Accordingly, H. pylori inside these yeasts can be carried by honeybees to different sugar- and nutrient-rich environments. Sugar-rich environments and honeybees play an important role in distribution of H. pylori-positive yeasts in nature.

Effect of different sanitizers against Zygosaccharomyces rouxii.

Microbial contamination is a problem in food industry. The effects produced by the metabolic activity of yeasts are diverse. If they are found in high numbers it might affect the organoleptic quality of the product. Zygosaccharomyces is the most frequent contaminant genus in sweetened juices that are not properly processed and stored. The aim of this work was to compare the action of five commercial sanitizers against isolates of Z. rouxii and to determine appropriate concentrations and time of action for the inactivation. Peracetic acid, monochloramine, iodophor and quaternary ammonium compounds were evaluated on different surfaces (stainless steel, plastic and glass) at room temperature for 30 and 60 s. It was possible to achieve a 99.99 of death Efficiency corresponding to 4 log10 of reduction for a contact time of 60 s for 0.5% (v/v) of peracetic acid, monochloramines and 0.5% (w/v) of iodophors, 1% for compound with a base of 8% (v/v) of benzyl chloride and 7.5% (v/v) of glutaraldehyde and 5% for sodium hypochlorite (55 g/L active compound). Stainless steel was successfully sanitized with all the compounds tested, and then it would be the most appropriate for the food industry because this material is also highly resistant to abrasion, to impact and to different chemical treatments. In view of the great variety of sanitizing products, the selection of the best sanitizer will depend on efficiency, safety and cost.

Discrimination of sweet wines partially fermented by two osmo-ethanol-tolerant yeasts by gas chromatographic analysis and electronic nose.

Some special sweet wines are obtained by partial fermentation of musts from off-vine dried grapes containing large amounts of sugars. This process is very slow and subject to serious stop problems that can be avoided by using osmo-ethanol-tolerant yeasts. Musts containing 371g/l of sugars were partially fermented with selected Saccharomyces cerevisiae strains, X4 and X5, to 12% (v/v) and the wines obtained with X5 exhibited a higher volatile acidity but lower concentrations of higher alcohols, carbonyl compounds and polyols than those obtained with X4. A principal component analysis (PCA) of the data provided by an electronic nose (E-nose) afforded discrimination between fermented and unfermented musts, but not between wines obtained with X4 or X5. The PCA applied to the major volatile compounds and polyols shows similar results, but a clear discrimination between wines is obtained by removing the polyols glycerol and 2,3-butanediol from the PCA.

Culture medium optimization for osmotolerant yeasts by use of a parallel fermenter system and rapid microbiological testing.

In the present study, a culture medium for qualitative detection of osmotolerant yeasts, named OM, was developed. For the development, culture media with different concentrations of glucose, fructose, potassium chloride and glycerin were analyzed in a Biolumix™ test incubator. Selectivity for osmotolerant yeasts was guaranteed by a water activity (aw)-value of 0.91. The best results regarding fast growth of Zygosaccharomyces rouxii (WH 1002) were achieved in a culture medium consisting of 45% glucose, 5% fructose and 0.5% yeast extract and in a medium with 30% glucose, 10% glycerin, 5% potassium chloride and 0.5% yeast extract. Substances to stimulate yeast fermentation rates were analyzed in a RAMOS® parallel fermenter system, enabling online measurement of the carbon dioxide transfer rate (CTR) in shaking flasks. Significant increases of the CTR was achieved by adding especially 0.1-0.2% ammonium salts ((NH4)2HPO4, (NH4)2SO4 or NH4NO3), 0.5% meat peptone and 1% malt extract. Detection times and the CTR of 23 food-borne yeast strains of the genera Zygosaccharomyces, Torulaspora, Schizosaccharomyces, Candida and Wickerhamomyces were analyzed in OM bouillon in comparison to the selective culture media YEG50, MYG50 and DG18 in the parallel fermenter system. The OM culture medium enabled the detection of 102CFU/g within a time period of 2-3days, depending on the analyzed yeast species. Compared with YEG50 and MYG50 the detection times could be reduced. As an example, W. anomalus (WH 1021) was detected after 124h in YEG50, 95.5h in MYG50 and 55h in OM bouillon. Compared to YEG50 the maximum CO2 transfer rates for Z. rouxii (WH 1001), T. delbrueckii (DSM 70526), S. pombe (DSM 70576) and W. anomalus (WH 1016) increased by a factor ≥2.6. Furthermore, enrichment cultures of inoculated high-sugar products in OM culture medium were analyzed in the Biolumix™ system. The results proved that detection times of 3days for Z. rouxii and T. delbrueckii can be realized by using OM in combination with the automated test system even if low initial counts (101CFU/g) are present in the products. In conclusion, the presented data suggest that the OM culture medium is appropriate for the enrichment of osmotolerant yeasts from high-sugar food products.

Characterization of Osmotolerant Yeasts and Yeast-Like Molds from Apple Orchards and Apple Juice Processing Plants in China and Investigation of Their Spoilage Potential.

Yeasts and yeast-like fungal isolates were recovered from apple orchards and apple juice processing plants located in the Shaanxi province of China. The strains were evaluated for osmotolerance by growing them in 50% (w/v) glucose. Of the strains tested, 66 were positive for osmotolerance and were subsequently identified by 26S or 5.8S-ITS ribosomal RNA (rRNA) gene sequencing. Physiological tests and RAPD-PCR analysis were performed to reveal the polymorphism of isolates belonging to the same species. Further, the spoilage potential of the 66 isolates was determining by evaluating their growth in 50% to 70% (w/v) glucose and measuring gas generation in 50% (w/v) glucose. Thirteen osmotolerant isolates representing 9 species were obtained from 10 apple orchards and 53 target isolates representing 19 species were recovered from 2 apple juice processing plants. In total, members of 14 genera and 23 species of osmotolerant isolates including yeast-like molds were recovered from all sources. The commonly recovered osmotolerant isolates belonged to Kluyveromyces marxianus, Hanseniaspora uvarum, Saccharomyces cerevisiae, Zygosaccharomyces rouxii, Candida tropicalis, and Pichia kudriavzevii. The polymorphism of isolates belonging to the same species was limited to 1 to 3 biotypes. The majority of species were capable of growing within a range of glucose concentration, similar to sugar concentrations found in apple juice products with a lag phase from 96 to 192 h. Overall, Z. rouxii was particularly the most tolerant to high glucose concentration with the shortest lag phase of 48 h in 70% (w/v) glucose and the fastest gas generation rate in 50% (w/v) glucose.