Evaluation of Foodborne Pathogen Die-off in Back-Sweetened Wine and Apple Cider Models.

ABSTRACT: Wine and alcoholic apple cider are commonly back-sweetened with unpasteurized juice to produce fresh, natural, and palatable sweetened alcoholic beverages. Foodborne pathogens may be introduced from unpasteurized juice into alcoholic beverages through this back-sweetening process. Although foodborne pathogens generally do not survive under low pH conditions or a high alcohol environment, the die-off of these pathogens has not been established to ensure the microbiological safety of the products. To establish the holding conditions that would provide the required 5-log pathogen reduction requirements for these back-sweetened beverages, we evaluated the survival of three common foodborne pathogens, E. coli O157:H7, Salmonella enterica, and Listeria monocytogenes, in modified white grape juice and apple juice models. White grape juice and apple juice were modified with hydrochloric acid and sodium hydroxide and with ethanol to achieve conditions that are similar to back-sweetened white wine and alcoholic apple cider in regard to pH and ethanol content. Foodborne pathogen cocktails were inoculated separately into modified juice models, and their survival in the juice models was recorded over a 96-h period. Our results show that a combination of low pH and high ethanol content resulted in faster pathogen die-off compared with higher pH and lower ethanol conditions. The holding times required for different combinations of pH and ethanol concentration for each juice model to achieve a 5-log reduction were reported. This research provides data to validate pathogen die-off to comply with juice hazard analysis and critical control point 5-log pathogen inactivation requirements for back-sweetened wine and alcoholic apple cider.

Genome rearrangements can make and break small RNA genes.

Small RNAs (sRNAs) are short, transcribed regulatory elements that are typically encoded in the intergenic regions (IGRs) of bacterial genomes. Several sRNAs, first recognized in Escherichia coli, are conserved among enteric bacteria, but because of the regulatory roles of sRNAs, differences in sRNA repertoires might be responsible for features that differentiate closely related species. We scanned the E. coli MG1655 and Salmonella enterica Typhimurium genomes for nonsyntenic IGRs as a potential source of uncharacterized, species-specific sRNAs and found that genome rearrangements have reconfigured several IGRs causing the disruption and formation of sRNAs. Within an IGR that is present in E. coli but was disrupted in Salmonella by a translocation event is an sRNA that is associated with the FNR/CRP global regulators and influences E. coli biofilm formation. A Salmonella-specific sRNA evolved de novo through point mutations that generated a σ(70) promoter sequence in an IGR that arose through genome rearrangement events. The differences in the sRNA pools among bacterial species have previously been ascribed to duplication, deletion, or horizontal acquisition. Here, we show that genomic rearrangements also contribute to this process by either disrupting sRNA-containing IGRs or creating IGRs in which novel sRNAs may evolve.

Mesoaciditoga lauensis gen. nov., sp. nov., a moderately thermoacidophilic member of the order Thermotogales from a deep-sea hydrothermal vent.

A novel moderately thermophilic, heterotrophic bacterium was isolated from a deep-sea hydrothermal vent deposit from the Mariner field along the Eastern Lau Spreading Center of the south-western Pacific Ocean. Cells were short motile rods (about 0.4×0.8 µm) that occurred singly or in pairs and were surrounded by a sheath-like membrane or 'toga'. The cells grew between 45 and 65 °C (optimum 57-60 °C) and at pH 4.1-6.0 (optimum pH 5.5-5.7) and grew optimally at 3 % (w/v) NaCl. The isolate grew on a range of carbon and proteinaceous substrates and reduced sulfur. The G+C content of the DNA was about 45 mol%. Phylogenetic analysis of the 16S rRNA gene sequence placed the new isolate as a deeply diverging lineage within the order Thermotogales. Based on the physiological, morphological and phylogenetic data, the isolate represents a novel species of a new genus with the proposed name Mesoaciditoga lauensis gen. nov., sp. nov. The type strain of Mesoaciditoga lauensis is cd-1655R(T) ( = DSM 25116(T) = OCM 1212(T)).