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In nature, Listeria may interact competitively and cooperatively with other organisms, resulting in unique spatial organization and functions for cells within the community. This study was undertaken to characterize the biofilm architecture of binary biofilms of Listeria monocytogenes and Lactobacillus species and to assess their effect on the survival of Listeria during exposure to hypochlorite. Three L. monocytogenes strains, ATCC 19115 (Lm5), ATCC 19117 (Lm7), and Coleslaw (LmC), were selected and combined individually with three Lactobacillus strains: L. fermentum (Lf), L. bavaricus (Lb), and L. plantarum (Lp). In binary Lm-Lp biofilms, the Lm cell counts were similar to single-species biofilms (8.5 log CFU/well), and the Lp cell numbers declined by 1.0 log CFU/well. In the presence of Lb, the Lm cell counts were reduced by 1.5 log CFU/well (p < 0.05), whereas the Lf cell counts increased at least by 3.5 log CFU/well. Confocal laser scanning microscopy (CLSM) determined that interspecies interactions significantly affected the spatial organization of three binary biofilms. Biofilm surface-to-volume ratio increased from 0.8 μm2/μm3 for Lm5 in the monoculture to 2.1 μm2/μm3 for Lm5-Lp in the dual-species model (p < 0.05), and was characterized by a thicker structure with a largely increased surface area. Biofilm roughness increased from 0.2 for Lm7 to 1.0 for Lm7-Lb biofilms (p < 0.05), which appeared as interspecific segregation. Biofilm thickness increased from 34.2 μm for LmC to 46.3 μm for LmC–Lf (p < 0.05), which produced flat and compact structures that covered the entire surface available. The biomass of the extracellular matrix was higher in the case of some binary biofilms (p < 0.05); however, this effect was dependent upon the species pair. When treated with hypochlorite, Lm5 in binary biofilms had an approximately 1.5 log CFU/well greater survival than individually. The unique spatial organization and greater protein production may explain the protective effect of Lp after hypochlorite exposure.

Salmonella and enterohemorrhagic Escherichia coli (EHEC) are of serious concern in wheat flour and its related products but little is known on their survival and thermal death kinetics. This study was undertaken to determine their long-term viability and thermal inactivation kinetics in flour. Inoculation was performed using mixtures of EHEC serogroups O45, O121, O145 and Salmonella followed by storage at room temperature (23°C) or 35°C (for Salmonella). Plate counting on tryptic soy agar (TSA) and enrichment were used to assess long-term survival. For thermal studies, wheat flour samples were heated at 55, 60, 65, and 70°C and cell counts of EHEC and Salmonella were determined by plating. The δ-values were calculated using the Weibull model. At room temperature, EHEC serovars and Salmonella were quantifiable for 84 and 112 days, and were detectable for the duration of the experiment after 168 and 365 days, respectively. The δ-values were 2.0, 5.54, and 9.3 days, for EHEC O121, O45, and O145, respectively, and 9.7 days for Salmonella. However, the only significant difference among all values was the δ-value for Salmonella and serogroup O121 (p ≤ 0.05). At 35°C, Salmonella counts declined to unquantifiable levels after a week and were not detected upon enrichment after 98 days. Heat treatment of inoculated wheat flour at 55, 60, 65, and 70°C resulted in δ-value ranges of 20.0–42.9, 4.9–10.0, 2.4–3.2, and 0.2–1.6 min, respectively, for EHEC. The δ-values for Salmonella at those temperatures were 152.2, 40.8, 17.9, and 17.4 min, respectively. The δ-values obtained for Salmonella at each temperature were significantly longer than for EHEC (p ≤ 0.05). Weibull model was a good fit to describe the thermal death kinetics of Salmonella and EHEC O45, O121 and O145 in wheat flour. HIGHLIGHTS -EHEC and Salmonella can survive for extended periods of time in wheat flour.-Long-term storage inactivation curves of EHEC and Salmonella were similar.-EHEC was more sensitive to heat than Salmonella.-Weibull model was a good fit to describe thermal death kinetics of EHEC and Salmonella.-Flour storage at 35°C may be a feasible method for microbial reduction.