Stevia-based sweeteners as a promising alternative to table sugar: The effect on Listeria monocytogenes and Salmonella Typhimurium growth dynamics.

Sugar is commonly substituted with stevia-based products in food industry and in our daily-life. This substitution results in a change in food product characteristic formula and properties that may affect the growth dynamics of food pathogenic and spoilage bacteria. This work studies the effect of table sugar (TS), laboratory sucrose (LS), commercial stevia (St) and steviol glycosides (SG) on the growth dynamics of Salmonella Typhimurium and Listeria monocytogenes. Experiments were carried out in general and minimal culture media at 3 equivalent concentration levels in terms of sweetness intensity (TS and LS at 3, 9 and 15% (w/v); St at 0.3, 0.9 and 1.5% (w/v); and SG at 0.01, 0.03 and 0.05% (w/v)). Incubation temperatures were: 4, 8 and 20°C for general media, and for minimal media 20°C. To decipher the role of these sweeteners, their concentration evolution in minimal media was determined via HPLC analysis. The results revealed slow maximum specific growth rates (µmax) of S. Typhimurium in general media with increasing concentrations of TS and LS at 20°C; and reduced maximum cell population (Nmax) at 8°C. The growth of L. monocytogenes in general culture media remains invariable independently of the sweetener added, except at 4°C. At this critical temperature, the presence of TS, LS and St seems to facilitate the growth of L. monocytogenes, presenting higher µmax values in comparison to SG and the control. Varying bacterial response to changes in media formulation suggests that further research is required, focusing on revealing the microbial dynamics in structured media, as well as in real food products.

Recent trends in non-invasive in situ techniques to monitor bacterial colonies in solid (model) food.

Planktonic cells typically found in liquid systems, are routinely used for building predictive models or assessing the efficacy of food preserving technologies. However, freely suspended cells often show different susceptibility to environmental hurdles than colony cells in solid matrices. Limited oxygen, water and nutrient availability, metabolite accumulation and physical constraints due to cell immobilization in the matrix, are main factors affecting cell growth. Moreover, intra- and inter-colony interactions, as a consequence of the initial microbial load in solid systems, may affect microbial physiology. Predictive food microbiology approaches are moving toward a more realistic resemblance to food products, performing studies in structured solid systems instead of liquids. Since structured systems promote microbial cells to become immobilized and grow as colonies, it is essential to study the colony behavior, not only for food safety assurance systems, but also for understanding cell physiology and optimizing food production processes in solid matrices. Traditionally, microbial dynamics in solid systems have been assessed with a macroscopic approach by applying invasive analytical techniques; for instance, viable plate counting, which yield information about overall population. In the last years, this approach is being substituted by more mechanistically inspired ones at mesoscopic (colony) and microscopic (cell) levels. Therefore, non-invasive and in situ monitoring is mandatory for a deeper insight into bacterial colony dynamics. Several methodologies that enable high-throughput data collection have been developed, such as microscopy-based techniques coupled with image analysis and OD-based measurements in microplate readers. This research paper provides an overview of non-invasive in situ techniques to monitor bacterial colonies in solid (model) food and emphasizes their advantages and inconveniences in terms of accuracy, performance and output information.

Effectiveness of chitosan against mature biofilms formed by food related bacteria.

Chitosan has proven antimicrobial properties against planktonic cell growth. Little is known, however, about its effects on already established biofilms. Oriented for application in food industry disinfection, the effectiveness of both medium molecular weight (MMW) chitosan and its enzymatically hydrolyzed product was tested against mature biofilms of four pathogenic strains, Listeria monocytogenes, Bacillus cereus, Staphylococcus aureus and Salmonella enterica, and a food spoilage species, Pseudomonas fluorescens. Unexpectedly, log reductions were in some cases higher for biofilm than for planktonic cells. One hour exposure to MMW chitosan (1% w/v) caused a 6 log viable cell reduction on L. monocytogenes monospecies mature biofilms and reduced significantly (3-5 log reductions) the attached population of the other organisms tested, except S. aureus. Pronase-treated chitosan was more effective than MMW chitosan on all tested microorganisms, also with the exception of S. aureus, offering best results (8 log units) against the attached cells of B. cereus. These treatments open a new possibility to fight against mature biofilms in the food industry.