Assessment of Viability of Listeria monocytogenes by Flow Cytometry.

In food industry, Listeria monocytogenes contamination can occur accidentally despite the quality control of raw materials and factory. Decontamination processes or inhibitory effects of ingredients/additives in food products are set up to ensure compliance with hygiene and microbiological criteria. These actions represent stresses for the pathogenic agent, causing fluctuations in its physiological states. Moreover, during these environmental stresses, Listeria monocytogenes can enter in a viable but nonculturable (VBNC) state which is not detected by plate counting but by flow cytometry. This technique coupled with cell staining by fluorescent dyes offers the possibility to assess different physiological states based on different cellular parameters: enzymatic activity, transmembrane integrity, membrane potential, and respiratory activity. In this chapter, we present a method to assess the viability of foodborne pathogens using a double-staining principle based on the assessment of membrane integrity and intracellular esterase activity.

Blackthorn fruit peel polyphenol extracts and photodynamic effect under blue light against Listeria monocytogenes.

Photodynamic inactivation is an emerging antimicrobial treatment that can be enhanced by employing exogenous photosensitizers to eradicate foodborne pathogens. This study investigated a novel combinatory strategy to eradicate Listeria monocytogenes using blackthorn fruit peel (BFP) and blue light (BL). Extracts of BFP were characterized in terms of polyphenolic content, individual constituents, and antioxidant and antimicrobial activity. The concentration of phenolic compounds and antioxidant activity were both found to be determinants of antimicrobial activity. It was further speculated that flavonols, predominantly quercetin and rutin, were responsible for the activity of BFP against L. monocytogenes. A combination of BFP and BL resulted in a rapid inactivation of the pathogen by up to 4 log CFU/mL at 58.5 J/cm2, corresponding to 15 min BL illumination. Flow cytometry analysis revealed that the bacterial cells lost activity and suffered extensive membrane damage, exceeding 90% of the population. After photosensitizing L. monocytogenes with the BFP constituents quercetin and rutin, a 1.3-log reduction was observed. When applied together, these compounds could inflict the same damaging effect on cells as they did individually when effects were added. Therefore, the results indicate that BFP represents a natural source of (pro-)photosensitizers, which act additively to create inactivation effects. This study may help identify more effective plant-based photosensitizers to control L. monocytogenes in food-related applications.

The Probiotic Enterococcus Lactis SF68 as a Potential Food Fermentation Microorganism for Safe Food Production.

Due to the reports describing virulent and multidrug resistant enterococci, their use has become a topic of controversy despite most of them being safe and commonly used in traditionally fermented foods worldwide. We have characterized Enterococcus lactis SF68, a probiotic strain approved by the European Food Safety Authority (EFSA) for use in food and feed, and find that it has a remarkable potential in food fermentations. Genome analysis revealed the potential of SF68 to metabolize a multitude of carbohydrates, including lactose and sucrose, which was substantiated experimentally. Bacteriocin biosynthesis clusters were identified and SF68 was found to display a strong inhibitory effect against Listeria monocytogenes. Fermentation-wise, E. lactis SF68 was remarkably like Lactococcus lactis and displayed a clear mixed-acid shift on slowly fermented sugars. SF68 could produce the butter aroma compounds, acetoin and diacetyl, the production of which was enhanced under aerated conditions in a strain deficient in lactate dehydrogenase activity. Overall, E. lactis SF68 was found to be versatile, with a broad carbohydrate utilization capacity, a capacity for producing bacteriocins, and an ability to grow at elevated temperatures. This is key to eliminating pathogenic and spoilage microorganisms that are frequently associated with fermented foods.

Interspecies Interaction of Bacteria Listeria monocytogenes, Yersinia pseudotuberculosis, and Marine Saprotrophs during Long-Term Culturing in Biofilms.

All bacterial strains studied retained the viability and ability to form both mono- and polycultural biofilms under conditions of long-term culturing in artificial seawater at 6°C and without addition of nutrients. Bacillus sp. and Pseudomonas japonica presumably stimulated the growth and reproduction of the pathogenic bacteria Listeria monocytogenes and Yersinia pseudotuberculosis. Preserved cell viability in a monoculture biofilm for a long period without adding a food source can indicate allolysis. At the same time, in a polycultural biofilm, the metabolites secreted by saprotrophic strains can stimulate the growth of L. monocytogenes and Y. pseudotuberculosis.

Antimicrobial resistance induction potential of grapefruit seed extract on multi-species biofilm of E. coli in food industry.

Biofilm formation in natural environments involving complex multi-structural arrangements hinders challenges in antimicrobial resistance. This study investigated the antimicrobial resistance potential of grapefruit seed extract (GSE) by examining the formation of mono-, dual-, and multi-species biofilms. We also explored the counterintuitive effect in response to GSE at various concentrations, including minimum inhibitory concentration (MIC) and sub-MIC (1/2 and 1/4 MIC). The results of the swimming and swarming motility tests revealed increased motility at the sub-MIC of GSE. The crystal violet assay demonstrated increased biofilm formation in multi-species biofilms, highlighting the synergistic effect of Escherichia coli, Salmonella Typhimurium, and Listeria monocytogenes. At the MIC concentration of GSE, field emission scanning electron microscopy (FE-SEM) revealed cell morphology damage, while sub-MIC increased biofilm formation and architectural complexity. Multi-species biofilms demonstrated greater biofilm-forming ability and antimicrobial resistance than mono-species biofilms, indicating synergistic interactions and enhanced resilience. These findings highlight the importance of understanding biofilm dynamics and antimicrobial resistance to ensure environmental safety.

Effect of Rheum ribes L. pulp enriched with eugenol or thymol on survival of foodborne pathogens and quality parameters of chicken breast fillets.

The aim of this study was to characterize the pulp of Rheum ribes L. and to determine the effect of the pulp enriched with eugenol (1 %) or thymol (1 %) on the microbiological and physico-chemical quality of chicken breast fillets. Chicken breast fillets, inoculated with Listeria monocytogenes, Salmonella enterica subsp. enterica serovar Typhimurium, and Escherichia coli O157:H7 (~6.0 log10), were marinated for 24 h in a mixture prepared from a combination of Rheum ribes L. pulp with eugenol or thymol. The quality parameters were analyzed for 15 days at +4 °C. The Rheum ribes L. pulp was found to have high antioxidant activity, high total phenolic content and contained 22 different phenolic substances, among which rutin ranked first. The pulp contained high levels of p-xylene and o-xylene as volatile substances and citric acid as an organic acid. The combination of Pulp + Eugenol + Thymol (PET) reduced the number of pathogens in chicken breast fillets by 2.03 to 3.50 log10 on day 0 and by 2.25 to 4.21 log10 on day 15, compared to the control group (P < 0.05). The marinating treatment significantly lowered the pH values of fillet samples on the first day of the study, compared to the control group (P < 0.05). During storage, TVB-N levels showed slower increase in the treatment groups compared to the control group (P < 0.05). In addition, the marinating process led to significant changes in physicochemical parameters such as water holding capacity, color, texture, cooking loss, and drip loss compared to the control group (P < 0.05). In conclusion, the results of this study showed that the pulp of Rheum ribes L., which has a high antioxidant capacity and contains various bioactive compounds. Furthermore, S. Typhimurium, E. coli O157:H7 and L. monocytogenes were inhibited considerably by marinating Rheum ribes L. pulp with a combination of eugenol and thymol.

Growth of Listeria monocytogenes in fresh vegetables and vegetable salad products: An update on influencing intrinsic and extrinsic factors.

The ability of foodborne pathogens to grow in food products increases the associated food safety risks. Listeria monocytogenes (Lm) is a highly adaptable pathogen that can survive and grow under a wide range of environmental circumstances, including otherwise inhibitory conditions, such as restrictive cold temperatures. It can also survive long periods under adverse environmental conditions. This review examines the experimental evidence available for the survival and growth of Lm on fresh vegetables and ready-to-eat vegetable salads. Published data indicate that, depending on certain intrinsic (e.g., nutrient composition) and extrinsic factors (e.g., storage temperature, packaging atmosphere), Lm can survive on and in a wide variety of vegetables and fresh-cut minimally processed vegetable salads. Studies have shown that temperature, modified atmosphere packaging, relative humidity, pH, water activity, background microbiota of vegetables, microbial strain peculiarities, and nutrient type and availability can significantly impact the fate of Lm in vegetables and vegetable salads. The influence of these factors can either promote its growth or decline. For example, some studies have shown that background microbiota inhibit the growth of Lm in vegetables and minimally processed vegetable salads, but others have reported a promoting, neutral, or insignificant effect on the growth of Lm. A review of relevant literature also indicated that the impact of most influencing factors is related to or interacts with other intrinsic or extrinsic factors. This literature synthesis contributes to the body of knowledge on possible strategies for improving food safety measures to minimize the risk of Lm-associated foodborne outbreaks involving vegetables and vegetable salads.

Formation and recovery of Listeria monocytogenes in viable but nonculturable state under different temperatures combined with low nutrition and high NaCl concentration.

The viable but nonculturable (VBNC) state occurs when bacteria lose their ability to grow and multiply on conventional media when stressed by adverse environmental factors, but they remain active and can revive under certain conditions, posing a food safety risk. In this study, the VBNC state of Listeria monocytogenes was induced with different temperatures combined with low nutrient conditions; the VBNC state of L. monocytogenes was confirmed in conjunction with the housekeeping gene abcZ using a molecular biology assay (PMA-qPCR) to calculate the viable bacterial count; The resuscitation conditions for the VBNC state of L. monocytogenes were investigated utilizing various nutrients in the culture medium and pasteurized milk. Four strains of L. monocytogenes reached the VBNC stage after 14, 21, 21, and 35 days at 20°C with 20% (or 30%) NaCl. Resuscitation studies indicate that Trypticase Soy Broth (TSB) combined with Tween 80 and sodium pyruvate is more effective for resuscitation. The Chinese national standard technology GB 4789.30-2016 was used to inoculate lettuce, chicken, and pasteurized milk with L. monocytogenes ATCC 19115 VBNC state. This research has significant implications for commercial food processing, long-term storage, disinfection, disease prevention, and control.

The combined effect of pH and NaCl on the susceptibility of Listeria monocytogenes to rhamnolipids.

The use of natural and sustainable additives, that are less aggressive to the environment, is a trend in the food industry. Rhamnolipids (RL) biosurfactants have shown potential for controlling food pathogens however, due to the presence of free carboxyl groups, the pH and ionic strength may influence the properties of such surfactants. In this study, we describe the antimicrobial activity of RL under different pH values and NaCl concentrations, towards both planktonic and biofilms of Listeria monocytogenes. RL were effective at pH 5.0 and the addition of 5 % NaCl improved the bactericidal efficacy for planktonic and sessile cells. The effect of NaCl was more pronounced at pH above 6 showing a significant increase in RL antimicrobial activity. At pH 7.0 planktonic population was eradicated by RL only when salt was present whereas biofilm viability was decreased by 5 log with MBIC varying from > 2500.0 mg/L (RL) to 39.0 mg/L (RL + 5 % NaCl). Larger vesicular and lamellar RL self-assembly structures were predominant when NaCl was present, suggesting their association with the antimicrobial activity observed. The pH and ionic strength of the medium are important parameters to be considered for the development of RL-based strategies to control L. monocytogenes.

Unrevealing the potentialities in food formulations of a low-branched dextran from Leuconostoc mesenteroides.

The search for novel exopolysaccharides (EPS) with targeted functionalities is currently a topic of great interest. This study aimed to investigate the chemical characteristics and technological properties of a novel EPS (named EPS_O) from Leuconostoc mesenteroides. EPS_O was a high-molecular-weight dextran (>6.68 × 105 g/mol) characterized by high water-holding capacity (785 ± 73%) and high water solubility index (about 99%). EPS_O in water (<30 mg/mL) formed viscous solutions, whereas at concentrations >30 mg/mL, it formed weak gels. Notably, lower concentrations (4-5 mg/mL) exhibited antimicrobial activity against various foodborne pathogens, antibiofilm activity against Listeria monocytogenes, and radical-scavenging activity. These properties are significant for maintaining food quality and promoting health. Based on these findings, EPS_O presents itself as a promising food ingredient that could elevate food quality and confer health benefits to consumers.

Antibacterial activities of Adina rubella extract enhanced by fermentation and its application in packaging films.

Food spoilage caused by pathogens pose great threat to food safety and human health. Plastarch-based packaging films with antibacterial activities provide an effective way to control foodborne pathogens. In this study, microbial fermentation dominated by yeast was used for the first time to increase the antibacterial activity of Adina rubella extract (ARE). The best antimicrobial effect of ARE was observed by fermentation for 9 days. The minimum inhibitory concentration of ARE against Listeria monocytogenes was 3.125 mg/mL. ARE destroyed the structure of the cell wall, increased the permeability of the cell membrane, led to the leakage of nucleic acids, and induced the change of ROS level, which caused cell death of Listeria monocytogenes. ARE-based biodegradable films were prepared and their performance in pork packaging application was evaluated. The films showed effective antimicrobial properties and showed great potential for the development of safe and sustainable food packaging films.

Effect of ripening time on the content of bioactive peptides and fatty acids profile of Artisanal Coalho cheese.

The present study aimed to investigate the influence of ripening on the physicochemical, microbiological aspects, and fatty acid profile of Artisanal Coalho Cheeses and to detect if there are peptides with bioactive potential in their composition. Artisanal Coalho Cheese samples were kindly provided by a dairy farm located in Brazil in the Rio Grande do Norte state. A completely randomized design was adopted, with four maturation periods (0, 30, 45, and 60 days). Physicochemical traits (pH, total solids, moisture, non-fat solids, fat in total solids, protein, ash, fatty acid profile) and microbiological characterization (Salmonella sp, Listeria monocytogenes, total and thermotolerant coliforms, Staphylococcus aureus) were analyzed on cheese samples. Additionally, assays were performed for antioxidant and antihypertensive bioactivity through ACE and antimicrobial inhibition of the peptides extracted from the samples. There was a linear increase in total solids and ash content and a decrease in moisture content with increasing maturation time. The matured cheese samples had a lower pH than fresh Artisanal Coalho Cheese. Twenty-seven fatty acids were identified in the cheeses: 15 saturated, 07 monounsaturated, and 05 polyunsaturated, with a linear reduction of essential fatty acids (n6 and n3) during maturation. The microbiological quality of the cheeses was satisfactory, with an absence of undesirable bacteria in 92% of the cheese samples. Water-soluble peptide fractions from all periods tested showed antioxidant and antihypertensive potential with ACE control, and the maturation process potentiated these capacities, with a decline in these activities observed at 60 days. The antimicrobial activity against Gram-positive and Gram-negative bacteria increased with maturation, reaching better results until 60 days. The maturation process on wooden planks in the periods of 30, 45, and 60 days allows the production of Artisanal Coalho Cheese of an innovative character, safe to consumers from the microbiological point of view, with differentiated physicochemical and functional characteristics and good quality of lipid fraction compared to fresh cheese, enabling the addition of value to the dairy chain.

Which domestic refrigerator temperatures in Europe? - Focus on shelf-life studies regarding Listeria monocytogenes (Lm) in ready-to-eat (RTE) foods.

Listeria monocytogenes (Lm) is a pathogenic bacteria able to grow at refrigerated temperatures, widely distributed in the environment. This bacteria is susceptible to contaminate various food products of which refrigerated ready-to-eat foods (RTEF) may pose a risk for public health. In Europe, food business operators (FBOs) shall ensure that foodstuffs comply with the relevant microbiological criteria set out in the Regulation (EC) N°2073/2005. Food safety criteria for Lm are defined in RTEF throughout their shelf-life. FBOs should implement studies to demonstrate that the concentration of Lm does not exceed 100 CFU/g at the end of the shelf-life, taking into account foreseeable conditions of distributions, storage and use, including the use by consumers. However, this last part of the cold chain for food products is the most difficult to capture and control. For this purpose, the European Union Reference Laboratory for Lm (EURL Lm) launched an inquiry to its National Reference Laboratory network and reviewed the scientific literature from 2002 to 2020. The outcomes were integrated in the technical guidance document of the EURL Lm to assess shelf-life of RTEF which resulted in the recommendation to use 10 °C as the reference temperature to simulate the reasonably foreseen storage conditions in domestic refrigerators.

The role of Listeria monocytogenes PstA in ß-lactam resistance requires the cytochrome bd oxidase activity.

c-di-AMP is an essential second messenger that binds and regulates several proteins of different functions within bacterial cells. Among those, PstA is a structurally conserved c-di-AMP-binding protein, but its function is largely unknown. PstA is structurally similar to PII signal transduction proteins, although it specifically binds c-di-AMP rather than other PII ligands such as ATP and α-ketoglutarate. In Listeria monocytogenes, we found that PstA increases ß-lactam susceptibility at normal and low c-di-AMP levels, but increases ß-lactam resistance upon c-di-AMP accumulation. Examining a PstA mutant defective for c-di-AMP binding, we found the apo form of PstA to be toxic for ß-lactam resistance, and the c-di-AMP-bound form to be beneficial. Intriguingly, a role for PstA in ß-lactam resistance is only prominent in aerobic cultures, and largely diminished under hypoxic conditions, suggesting that PstA function is linked to aerobic metabolism. However, PstA does not control aerobic growth rate, and has a modest influence on the tricarboxylic acid cycle and membrane potential-an indicator of cellular respiration. The regulatory role of PstA in ß-lactam resistance is unrelated to reactive oxygen species or oxidative stress. Interestingly, during aerobic growth, PstA function requires the cytochrome bd oxidase (CydAB), a component of the respiratory electron transport chain. The requirement for CydAB might be related to its function in maintaining a membrane potential, or redox stress response activities. Altogether, we propose a model in which apo-PstA diminishes ß-lactam resistance by interacting with an effector protein, and this activity can be countered by c-di-AMP binding or a by-product of redox stress. IMPORTANCE: PstA is a structurally conserved c-di-AMP-binding protein that is broadly present among Firmicutes bacteria. Furthermore, PstA binds c-di-AMP at high affinity and specificity, indicating an important role in the c-di-AMP signaling network. However, the molecular function of PstA remains elusive. Our findings reveal contrasting roles of PstA in ß-lactam resistance depending on c-di-AMP-binding status. We also define physiological conditions for PstA function during aerobic growth. Future efforts can exploit these conditions to identify PstA interaction partners under ß-lactam stress.

Zein-based nisin-loaded electrospun nanofibers as active packaging mats for control of Listeria monocytogenes on peach.

Zein-based nanofibers (NFs) functionalized with nisin (NS), reinforced with montmorillonite nanoclay (nMMT) were fabricated by uniaxial electrospinning (ES) for the first time to preserve yellow peach. Spinnability/viscosity/conductivity optimizations generated porous (95.09%), bead-free, ultrathin (119 nm) NFs of low hydrophobicity (26.05°). Glutaraldehyde (GTA) crosslinking fostered positive outcomes of tensile strength (1.23 MPa), elongation (5.0%), hydrophobicity (99.46°), surface area (201.38 m2.g-1), pore size (2.88 nm), thermal stability (Tmax = 342 °C), antioxidant/cytotoxic activities in optimized NFs that released NS sustainably according to Korsmeyer-Peppas model indicating a Fickian diffusion mechanism with R2 = 0.9587. The novel NFs inhibited growth of Listeria monocytogenes/aerobic mesophilic populations in peach after 4 days of abusive storage, evincing their robustness in food contact applications. Simultaneously, quality parameters (moisture/texture/browning/total soluble solids/pH) and peach physical appearance were maintained for up to 8 days, endorsing the practical value of zein-based NFs as a non-thermal postharvest intervention for prolonging fruits storage life.

Disinfectant efficacy of glabridin against dried and biofilm cells of Listeria monocytogenes and the impact of residual organic matter.

Glabridin is an antimicrobial compound which can be extracted from plants, such as liquorice (Glycyrrhiza glabra) roots. Although its activity against foodborne pathogens and spoilage microorganisms has already been reported, the investigation of potential applications as a surface disinfectant is still largely unexplored. Hence, this study evaluated the disinfectant efficacy of glabridin against Listeria monocytogenes. The activity of glabridin was first tested in vitro in a nutrient-rich medium against eight strains of L. monocytogenes, including food isolates and the model strain EGDe. The tested strains showed similar susceptibility with minimal inhibitory and bactericidal concentrations of 12.5 µg/mL and 25 µg/mL, respectively. Subsequently, L. monocytogenes L6, FBR17 and EGDe were selected to assess the efficacy of glabridin against dried cells (according to the European standard EN 13697:2015 + A1:2019) and biofilm cells on stainless steel surfaces. Moreover, the impact of food residual organic matter was investigated using skim milk, cantaloupe and smoked salmon solution as soiling components. Our results showed that applying 200 µg/mL of glabridin resulted in a substantial reduction (>3 log10) of dried and biofilm cells of L. monocytogenes in standard conditions (i.e. low level of residual organic matter). Cantaloupe soiling components slightly reduced the activity of glabridin, while the efficacy of glabridin when tested with salmon and skim milk residuals was substantially affected. Comparative analysis using standardized protein contents provided evidence that the type of food matrices and type of proteins may impact the activity of glabridin as a disinfectant. Overall, this study showed low strain variability for the activity of glabridin against L. monocytogenes and shed light on the possible application of this natural antimicrobial compound as a surface disinfectant.

Modelling the colony growth dynamics of Listeria monocytogenes single cells after exposure to peracetic acid and acidic conditions.

Studies of classical microbiology rely on the average behaviour of large cell populations without considering that clonal bacterial populations may bifurcate into phenotypic distinct sub-populations by random switching mechanisms.Listeria monocytogenes exposure to sublethal stresses may induce different physiological states that co-exist (i.e., sublethal injury or dormancy) and present variable resuscitation capacity. Exposures to peracetic acid (PAA; 10-30 ppm; for 3 h), acetic acid and hydrochloric acid (AA and HCl; pH 3.0-2.5; for 5 h) at 20 °C were used to induce different physiological states in L. monocytogenes, Scott A strain. After stress exposure, colony growth of single cells was monitored, on Tryptic Soy Agar supplemented with 0.6 % Yeast Extract, using time-lapse microscopy, at 37 °C. Images were acquired every 5 min and were analyzed using BaSCA framework. Most of the obtained growth curves of the colonies were fitted to the model of Baranyi and Roberts for the estimation of lag time (λ) and maximum specific growth rate (µmax), except the ones obtained after exposure to AA pH 2.7 and 2.5 that were fitted to the Trilinear model. The data of λ and µmax that followed a multivariate normal distribution were used to predict growth variability using Monte Carlo simulations. Outgrowth kinetics after treatment with AA (pH 2.7 and 2.5; for 5 h at 20 °C), PAA (30 ppm; for 3 h at 20 °C) revealed that these stress conditions increase the skewness of the variability distributions to the right, meaning that the variability in lag times increases in favour of longer outgrowth. Exposures to AA pH 2.5 and 30 ppm PAA resulted in two distinct subpopulations per generation with different growth dynamics. This switching mechanism may have evolved as a survival strategy for L. monocytogenes cells, maximizing the chances of survival. Simulation of microbial growth showed that heterogeneity in growth dynamics is increased when cells are recovering from exposure to sublethal stresses (i.e. PAA and acidic conditions) that may induce injury or dormancy.

Application of rosemary oil nano-emulsion as antimicrobial and antioxidant natural alternative in pasteurized cream and Karish cheese.

Essential oils possess significant antimicrobial and antioxidant properties and are increasingly used as natural substitutes for food preservation. Therefore, this study investigated the potential application of rosemary essential oil (REO) and REO nano-emulsion in the dairy plant. The antimicrobial effects of REO and REO nano-emulsion were determined by an agar well diffusion assay after chemical profiling by Gas Chromatography-Mass Spectrometry (GC-MS). The REO nano-emulsion was characterized by a Transmission Electron Microscope (TEM). The REO chemical profile revealed the presence of 42 chemical compounds, including 1, 8-cineole (9.72 %), and α-pinene (5.46 %) as major active components. REO nano-emulsion demonstrated significant antimicrobial activity compared to REO (P < 0.05) with a MIC value of 0.0001 mg/ml against Listeria monocytogenes and Aspergillus flavus and 0.001 mg/ml against Pseudomonas aeruginosa and Bacillus cereus. REO nano-emulsion enhanced the oxidative stability of pasteurized fresh cream, revealing a non-significant difference compared with that inoculated with butylated hydroxy anisol (BHA; synthetic antioxidant) (P˃ 0.05). Fortified cream and Karish cheese with REO nano-emulsion were evaluated organoleptically, and the results showed higher grades of overall acceptability when compared to control samples with a statistically significant difference (P < 0.05). Viability studies were estimated using the previously mentioned microorganisms in fortified fresh cream and Karish cheese with REO nano-emulsion. Results of the fortified cream showed a complete reduction of L. monocytogenes, A. flavus, and B. cereus on days 5, 7, and 10, respectively, and a 96.93 % reduction of P. aeruginosa by the end of the storage period. Regarding Karish cheese viability studies, C. albicans, A. flavus, and P. aeruginosa exhibited complete reduction on days 10, 10, and 15 of storage, respectively. In conclusion, REO nano-emulsion was recommended as a natural, safe, and effective antimicrobial and antioxidant additive in the dairy industry.

Influence of slope, material, and temperature on Listeria monocytogenes and Pseudomonas aeruginosa mono- and dual-species biofilms.

Understanding factors influencing Listeria monocytogenes biofilms aid in developing more effective elimination/prevention strategies. This study examined the effect of temperature (4 °C, 21 °C, 30 °C), materials (stainless steel 316 L with 2B and 2 R finishes, glass, and polypropylene), and slope (0°/horizontal or 90°/vertical) on mono- and dual-species biofilms using two L. monocytogenes strains and one Pseudomonas aeruginosa strain. All biofilms were grown in 10% TSB for 24 h and analyzed using culture-based methods. Additionally, the architecture of monospecies biofilms was studied using fluorescence microscopy. Overall, P. aeruginosa showed higher biofilm formation potential (6.2 log CFU/cm2) than L. monocytogenes (4.0 log CFU/cm2). Temperature greatly influenced P. aeruginosa and varied for L. monocytogenes. The slope predominantly influenced L. monocytogenes monospecies biofilms, with cell counts increasing by up to 2 log CFU/cm2. Surface material had little impact on biofilm formation. The study highlights the varying effects of different parameters on multispecies biofilms and the importance of surface geometry.

Surface material does not affect L. monocytogenes or P. aeruginosa biofilmsL. monocytogenes produces more biofilm on horizontal surfaces than on vertical surfacesIn dual-species biofilm, L. monocytogenes shows enhanced biofilm productionIncreased temperature often results in enhanced biofilm formation.

Curcumin-based residue-free and reusable photodynamic inactivation system for liquid foods and its application in freshly squeezed orange juice.

To enhance curcumin's application in photodynamic inactivation (PDI) of liquid foods, a supramolecular complex of biotin-modified ß-cyclodextrin and curcumin (Biotin-CD@Cur) was synthesized. This complex significantly improves curcumin's solubility, stability, and PDI efficiency. Following PDI, Biotin-CD@Cur can be magnetically separated from the liquid matrix using streptavidin-coated magnetic beads (SA-MBs). Leveraging the reversible binding between streptavidin and biotin, Biotin-CD@Cur and SA-MBs fully dissociate in ultrapure water at 70 °C, enabling reuse. Antibacterial tests in freshly squeezed orange juice demonstrated that a low dose of 1.5 J/cm2 from a 420 nm LED array and 10 µg/mL of Biotin-CD@Cur achieved log reductions of 3.287 ± 0.015 for Staphylococcus aureus and 2.961 ± 0.011 for Listeria monocytogenes, while preserving the juice's flavor and nutritional contents. The PDI system remained effective for at least four cycles. Ultra-performance liquid chromatography and atomic absorption spectroscopy confirmed no residues of system components in the juice after magnetic separation.