The Development of a Digital Twin to Improve the Quality and Safety Issues of Cambodian Pâté: The Application of 915 MHz Microwave Cooking.

Foodborne diseases are common in Cambodia and developing good food hygiene practices is a mandatory goal. Moreover, developing a low-carbon strategy and energy efficiency is also a priority. This study focuses on pâté cooking, a very common food product in Cambodia. In this paper, the authors chose to develop a digital twin dedicated to perfectly predict the temperature for cooking in a 915 MHz single-mode cavity, instead of using a classical and energy-consuming steaming method. The heating strategy is based on a ramp-up heating and a temperature-holding technique (with Tylose® as the model food and Cambodian pâté). The model developed with COMSOL® Multiphysics software can accurately predict both local temperatures and global moisture losses within the pâté sample (RMSE values of 2.83 and 0.58, respectively). The moisture losses of Cambodian pâté at the end of the process was 28.5% d.b (dry basis) after a ramp-up heating activity ranging from 4 to 80 °C for 1880 s and a temperature-holding phase at 80 °C for 30 min. Overall, the accurate prediction of local temperatures within Cambodian pâté is mainly dependent on the external heat-transfer coefficient during the temperature-holding phase, and is specifically discussed in this study. A 3D model can be used, at present, as a digital twin to improve the temperature homogeneity of modulated microwave power inputs in the future.

Detection and genetic analysis of Escherichia coli from Tonle Sap Lake and its tributaries in Cambodia: Spatial distribution, seasonal variation, pathogenicity, and antimicrobial resistance.

As an indicator of fecal contamination, Escherichia coli was monitored in Tonle Sap Lake, Cambodia, and its tributaries during low- and high-water seasons, focusing on the impacts on floating villagers inhabiting boathouses. E. coli concentrations in the floating villages (3.6 × 103 and 5.7 × 103 CFU/100 mL during the low- and high-water seasons, respectively) were significantly higher than those in other lake sites (4.0 × 101 and 7.0 × 100 CFU/100 mL during the low- and high-water seasons, respectively) and rivers (3.3 × 102 and 8.9 × 102 CFU/100 mL during the low- and high-water seasons, respectively), most likely because fecal materials from the boathouses were discharged without treatment. At most of the lake sampling sites remote from the boathouses, the E. coli concentration was lower during the high-water season than that during the low-water season, due to dilution by lake water. E. coli colonies detected during monitoring were isolated for pathotyping, antimicrobial susceptibility testing, beta-lactamase gene detection, and multilocus sequencing typing (MLST). Of the 659 E. coli isolates, 101 (15.3%) were diarrheagenic E. coli (DEC). The prevalence of DEC (52.2%) in the floating villages during the low-water season was higher than that during the high-water season (4.2%) and that in other sites during both seasons (10.6-21.3%). The DEC isolates from the floating villages during the low-water season showed high antimicrobial resistance, including ampicillin (83.4%) and ciprofloxacin (83.4%), and frequently possessed a beta-lactamase gene (blaTEM) (83.4%). MLST analysis indicated that the predominant sequence type (ST) of DEC isolates from the floating villages possibly originated from humans, whereas more diverse STs were detected in isolates from other sites. We revealed the wide presence of diarrheagenic and antimicrobial-resistant E. coli in Tonle Sap Lake and identified a considerable infection risk in floating villages, especially during the low-water season.

Engineering Synthetic Microbial Communities through a Selective Biofilm Cultivation Device for the Production of Fermented Beverages.

Production of Cambodian rice wine involves complex microbial consortia. Indeed, previous studies focused on traditional microbial starters used for this product revealed that three microbial strains with complementary metabolic activities are required for an effective fermentation, i.e., filamentous fungi (Rhizopus oryzae), yeast (Saccharomyces cerevisiae), and lactic acid bacteria (Lactobacillus plantarum). Modulating the ratio between these three key players led to significant differences, not only in terms of ethanol and organic acid production, but also on the profile of volatile compounds, in comparison with natural communities. However, we observed that using an equal ratio of spores/cells of the three microbial strains during inoculation led to flavor profile and ethanol yield close to that obtained through the use of natural communities. Compartmentalization of metabolic tasks through the use of a biofilm cultivation device allows further improvement of the whole fermentation process, notably by increasing the amount of key components of the aroma profile of the fermented beverage (i.e., mainly phenylethyl alcohol, isobutyl alcohol, isoamyl alcohol, and 2-methyl-butanol) and reducing the amount of off-flavor compounds. This study is a step forward in our understanding of interkingdom microbial interactions with strong application potential in food biotechnology.

Prospects for Food Fermentation in South-East Asia, Topics From the Tropical Fermentation and Biotechnology Network at the End of the AsiFood Erasmus+Project.

Fermentation has been used for centuries to produce food in South-East Asia and some foods of this region are famous in the whole world. However, in the twenty first century, issues like food safety and quality must be addressed in a world changing from local business to globalization. In Western countries, the answer to these questions has been made through hygienisation, generalization of the use of starters, specialization of agriculture and use of long-distance transportation. This may have resulted in a loss in the taste and typicity of the products, in an extensive use of antibiotics and other chemicals and eventually, in a loss in the confidence of consumers to the products. The challenges awaiting fermentation in South-East Asia are thus to improve safety and quality in a sustainable system producing tasty and typical fermented products and valorising by-products. At the end of the "AsiFood Erasmus+ project" (www.asifood.org), the goal of this paper is to present and discuss these challenges as addressed by the Tropical Fermentation Network, a group of researchers from universities, research centers and companies in Asia and Europe. This paper presents current actions and prospects on hygienic, environmental, sensorial and nutritional qualities of traditional fermented food including screening of functional bacteria and starters, food safety strategies, research for new antimicrobial compounds, development of more sustainable fermentations and valorisation of by-products. A specificity of this network is also the multidisciplinary approach dealing with microbiology, food, chemical, sensorial, and genetic analyses, biotechnology, food supply chain, consumers and ethnology.

Impact of Microbial Composition of Cambodian Traditional Dried Starters (Dombea) on Flavor Compounds of Rice Wine: Combining Amplicon Sequencing With HP-SPME-GCMS.

Dombae is a traditional ferment starter which has been used for starchy based wine production in Cambodia. However, the production technology of rice wine in Cambodia is not optimized. The current study aimed to investigate the microbiota associated in five ferment starters and the effect of a traditional fermentation process using a metagenomics sequencing analysis and HS-SPME-GCMS for the characterization of the aromatic profiles at the end of fermentation. Most of bacteria identified in this study were lactic acid bacteria including Weissella cibaria, Pediococcus sp. MMZ60A, Lactobacillus fermentum, and Lactobacillus plantarum. Saccharomyces cerevisiae and Saccharomycopsis fibuligera were found to be abundant yeasts while the only amylolytic filamentous fungus was Rhizopus oryzae. A total of 25 aromatic compounds were detected and identified as esters, alcohols, acids, ketones and aldehydes. The alcohol group was dominant in each rice wine. Significant changes were observed at the level of microbial communities during fermentation, suggesting microbial succession for the assimilation of starch and subsequently assimilation of fermentation by-products leading to the production of flavor compounds. At this level, the presence of Weissella, Pediococcus, and Lactobacillus genus was strongly correlated with most of the flavor compounds detected.

Fate of Escherichia coli in dialysis device exposed into sewage influent and activated sludge.

Tracing the fate of pathogens in environmental water, particularly in wastewater, with a suitable methodology is a demanding task. We investigated the fate of Escherichia coli K12 in sewage influent and activated sludge using a novel approach that involves the application of a biologically stable dialysis device. The ion concentrations inside the device could reach that of surrounding solution when it was incubated in phosphate buffered saline for 2 h. E. coli K12 above 107 CFU mL-1 (inoculated in distilled water, influent, activated sludge) were introduced into the device and incubated in influent and activated sludge for 10 days. Without indigenous microorganisms, E. coli K12 could survive even with the limited ions and nutrients concentrations in influent and activated sludge. E. coli K12 abundance in influent and activated sludge were reduced by 60 and 85%, respectively, after just 1 day. The establishment of microbial community in wastewater played an important role in reducing E. coli K12. Bacteriophage propagated in filtered influent or activated sludge when E. coli K12 was introduced, but not in raw influent or activated sludge. The methodology developed in this study can be applied in the actual environmental water to trace the fate of pathogens.

The impact of oregano (Origanum heracleoticum) essential oil and carvacrol on virulence gene transcription by Escherichia coli O157:H7.

The aim of the current study was to determine, via reverse transcription quantitative polymerase chain reaction (RT-qPCR) analysis, the effect of oregano essential oil (Origanum heracleoticum) and carvacrol, its major component, on the expression of virulence-associated genes in enterohaemorrhagic Escherichia coli (EHEC) O157:H7 ATCC strain 35150. Both oregano oil and carvacrol demonstrated their efficacy firstly, by inhibiting the transcription of the ler gene involved in upregulation of the LEE2, LEE3 and LEE4 promoters and of attaching and effacing lesions and secondly by decreasing both Shiga toxin and fliC genes expression. In addition, a decrease in luxS gene transcription involved in quorum sensing was observed. These results were dose dependent and showed a specific effect of O. heracleoticum and carvacrol in downregulating the expression of virulence genes in EHEC O157:H7. These findings suggest that oregano oil and carvacrol have the potential to mitigate the adverse health effects caused by virulence gene expression in EHEC O157:H7, through the use of these substances as natural antibacterial additives in foods or as an alternative to antibiotics.

Antimicrobial activities of commercial essential oils and their components against food-borne pathogens and food spoilage bacteria.

This study was undertaken to determine the in vitro antimicrobial activities of 15 commercial essential oils and their main components in order to pre-select candidates for potential application in highly perishable food preservation. The antibacterial effects against food-borne pathogenic bacteria (Listeria monocytogenes, Salmonella Typhimurium, and enterohemorrhagic Escherichia coli O157:H7) and food spoilage bacteria (Brochothrix thermosphacta and Pseudomonas fluorescens) were tested using paper disk diffusion method, followed by determination of minimum inhibitory (MIC) and bactericidal (MBC) concentrations. Most of the tested essential oils exhibited antimicrobial activity against all tested bacteria, except galangal oil. The essential oils of cinnamon, oregano, and thyme showed strong antimicrobial activities with MIC ≥ 0.125 µL/mL and MBC ≥ 0.25 µL/mL. Among tested bacteria, P. fluorescens was the most resistant to selected essential oils with MICs and MBCs of 1 µL/mL. The results suggest that the activity of the essential oils of cinnamon, oregano, thyme, and clove can be attributed to the existence mostly of cinnamaldehyde, carvacrol, thymol, and eugenol, which appear to possess similar activities against all the tested bacteria. These materials could be served as an important natural alternative to prevent bacterial growth in food products.