Predicting the impact of temperature and relative humidity on Salmonella growth and survival in sliced chard, broccoli and red cabbage.

This study assessed the fate of a Salmonella enterica cocktail (S. Typhimurium, S. Enteritidis, S. Newport, S. Agona and S. Anatum; initial counts 3.5 log CFU/g) in minimally processed sliced chard, broccoli and red cabbage at 16 conditions of different temperature (7, 14, 21 and 37 °C) and relative humidity (RH; 15, 35, 65 and 95%) over six days (144 h). Linear regression was used to estimate the rate change of Salmonella in cut vegetables as a function of temperature and relative humidity (RH). R2 value of 0.85, 0.87, and 0.78 were observed for the rates of change in chard, broccoli, and red cabbage, respectively. The interaction between temperature and RH was significant in all sliced vegetables. Higher temperatures and RH values favored Salmonella growth. As temperature or RH decreased, the rate of S. enterica change varied by vegetable. The models developed here can improve risk management of Salmonella in fresh cut vegetables.

Ozone and photodynamic inactivation of norovirus surrogate bacteriophage MS2 in fresh Brazilian berries and surfaces.

This study assessed the efficacy of ozone (bubble diffusion in water; 6.25 ppm) and photodynamic inactivation (PDT) using curcumin (75 µM) as photosensitizer (LED emission 430-470 nm; 33.6 mW/cm2 irradiance; 16.1, 20.2, and 24.2 J/cm2 light dose) against the Norovirus surrogate bacteriophage MS2 in Brazilian berries (black mulberry and pitanga) and surfaces (glass and stainless steel). Contaminated berries and surfaces were immersed in ozonized water or exposed to PDT-curcumin for different time intervals. Transmission electron microscopy was used to assess the effects of the treatments on MS2 viral particles. The MS2 inactivation by ozone and PDT-curcumin varied with the fruit and the surface tested. Ozone reduced the MS2 titer up to 3.6 log PFU/g in black mulberry and 4.1 log PFU/g in pitanga. On surfaces, the MS2 reduction by ozone reached 3.6 and 4.8 log PFU/cm2 on glass and stainless steel, respectively. PDT-curcumin reduced the MS2 3.2 and 4.8 log PFU/g in black mulberry and pitanga and 2.7 and 3.3 log PFU/cm2 on glass and stainless steel, respectively. MS2 particles were disintegrated by exposure of MS2 to ozone and PDT-curcumin on pitanga. Results can contribute to establishing effective practices for controlling NoV in fruits and surfaces, estimated based on MS2 bacteriophage behavior.

Survival kinetics, membrane integrity and metabolic activity of Salmonella enterica in conventionally and osmotically dehydrated coconut flakes.

Many outbreaks involving Salmonella enterica in dehydrated coconut have been reported. Little is known about the survival of S. enterica in dehydrated coconut flakes at common retail or domestic storage conditions. This study evaluated the behavior of a S. enterica cocktail (S. Enteritidis PT4, S. Typhimurium PT4, S. Bredeney, S. Muenster and S. Agona) in conventionally and osmotically dehydrated coconut flakes under four storage regimes: 25 °C for 120 days, 25 °C for 30 days then 7 °C for 90 days, 7 °C for 30 days then 25 °C for 90 days, and 7 °C for 120 days. S. enterica membrane integrity (using with propidium iodide and bis-1,3-dibutylbarbutyric acid) and metabolic activity (using 5-cyano-2,3-ditolyl tetrazolium chloride) were assessed by flow cytometry analysis after dehydration and storage at 7 °C or 25 °C for 120 days. Lower S. enterica inactivation rates (kmax 0.02 to 0.04 1/days) were observed in conventionally dehydrated coconut flakes compared to osmotically dehydrated coconut flakes (kmax 0.16 to 0.20 1/days). Changes in storage temperature did not affect the behavior of S. enterica in conventionally or osmotically dehydrated coconut flakes. Results show that S. enterica inactivation in conventionally dehydrated coconut flakes could be described by log-linear with tail models. S. enterica inactivation in osmotically dehydrated coconut flakes could be described by log-linear with shoulder and tail models. Subpopulations of S. enterica cells with damaged membranes and without metabolic activity were larger in conventionally (32.1% and 90.9%, respectively) than osmotically dehydrated coconut (18.5% and 82.2%, respectively) flakes at the beginning of the storage. Subpopulations of S. enterica cells with damaged membrane decreased by 9.4-14.4%, while cells with membrane potential and intact membrane increased by 23.7 and 24.2% in conventionally dehydrated coconut flakes after 120 days of storage at 7 °C or 25 °C, respectively. Subpopulations of S. enterica with damaged membranes did not change significantly in osmotically dehydrated coconut flakes. Our findings suggest that S. enterica populations decline during storage occurs due in part to membrane integrity losses. These data can contribute to the development of risk management strategies for S. enterica in dehydrated coconut flakes.

Genomic investigation of antimicrobial resistance determinants and virulence factors in Salmonella enterica serovars isolated from contaminated food and human stool samples in Brazil.

This study investigated the antimicrobial resistance determinants, virulence factors and identified serovars in 37 Salmonella enterica strains isolated from human stool and contaminated foods linked to outbreaks that occurred in Brazil over 7 years using whole genome sequencing (WGS). Phylogenetic analysis of selected serovars (S. Typhimurium, S. Infantis, S. London, and S. Johannesburg) was performed. Ten distinct serovars were identified and, 51% of the tested strains (n = 19) showed disagreement with the previous conventional serotyping. The antimicrobial resistance (AMR) determinants or plasmids varied among the strains. Resistome analysis revealed the presence of resistance genes to aminoglycosides [aac (6')-laa, aph (3″)-lb, aph (6)-ld, aadA1 and aadA2], sulfonamides (sul1), trimethoprin (dfrA8), fosfomycin (fosA7) and tetracyclines (tetA, tetB, tetC), as well as point mutations in parC (T57S) and gyrA (S83F). Plasmidome showed the presence of IncHI2, IncHI2A, IncFIB (S), IncFII (S), IncI1 and p0111 plasmids. Eight Salmonella pathogenicity islands and up to 102 stress and/or virulence genes were identified in the evaluated genomes. Virulence genes of K88 fimbrial adhesin were first reported in S. enterica (S. Pomona, S. Bredeney and S. Mbandaka strains). pilW gene was first identified in S. Pomona. Phylogenetic analysis showed that some serovars circulated in Brazil for decades, primarily within the poultry production chain. Findings highlighted the virulence and AMR determinants in strains that may lead to recurring food outbreaks.

Exploiting the use of agro-industrial residues from fruit and vegetables as alternative microalgae culture medium.

There is a need for searching new microalgae species, and the most suitable strategy to increase the cost-effectiveness of a microalgae culture system is to use resources of low costs, such as residues. This study aimed to evaluate the cultivation of microalgae isolated from the Brazilian Northeast region (Lagerheimia longiseta, Monoraphidium contortum, and Scenedesmus quadricauda) in an alternative medium of low cost (biocompost of discarded fruits and vegetables) with a view to possible applications in the food industry. Microalgae cultivated in the conventional synthetic medium was used as control. The cultivation of microalgae in the alternative medium allowed suitable cell growth, and improved the antioxidant activity and the levels of monounsaturated fatty acid and polyunsaturated fatty acid compared to the synthetic medium. The cultivation of S. quadricauda and L. longiseta species in the alternative medium resulted in increased protein content and/or total phenolic content, and improved health indices (lower levels of atherogenic, thrombogenic, and hypercholesterolemic saturated fatty acids indices, and higher levels of desired fatty acids index) compared to cultivation in synthetic medium. The cultivation of M. contortum in the alternative medium contributed to the production of higher lipid content, mainly saturated fatty acid (palmitic acid), which contributed negatively to the health indices. This study proved that S. quadricauda and L. longiseta microalga species from freshwaters have significant potential for distinct applications in functional food industries, and the biocompost of discarded fruits and vegetables is a suitable medium for microalgae cultivation.

Physiological alterations involved in inactivation of autochthonous spoilage bacteria in orange juice caused by Citrus essential oils and mild heat.

This study investigated physiological alterations involved in the inactivation of Levilactobacillus (L.) brevis and Leuconostoc (Lc.) mesenteroides in orange juice caused by Citrus lemon essential oil (CLEO) and C. reticulata essential oil (CREO) alone and combined with mild heat treatment (MHT). Damage in DNA, membrane integrity, membrane potential, metabolic and efflux activity of bacterial cells were measured after exposure (6 and 12 min) to CLEO or CREO (0.5 µL/mL) and/or MHT (54 °C) using flow cytometry. Limonene was the major constituent in CLEO (66.4%) and CREO (89.4%). The size of the damaged cell subpopulations increased (p < 0.05) after longer exposure time and varied with the tested essential oil and/or bacterial isolate. After exposure to CLEO and CREO alone, the cell subpopulations with damage in measured physiological functions were in a range of 19.6-66.8% and 23.8-75.9%, respectively. Exposure to CREO resulted in larger Lc. mesenteroides cell subpopulations (35.4-68.7%) with damaged DNA, permeabilized and depolarized membrane and compromised metabolic or efflux activity compared to L. brevis (23.8-58.0%). In contrast, exposure to CLEO led to higher damaged L. brevis cell subpopulations (35.1-77%) compared to Lc. mesenteroides (25.3-36.6%). Exposure to combined treatments (CLEO or CREO and MHT) affected the measured physiological functions in almost the entire L. brevis and Lc. mesenteroides cell population (up to 99%), although the damage extension on each isolate varied with tested essential oil. Results show that inactivation of L. brevis and Lc. mesenteroides cells caused by CLEO and CREO alone and combined with MHT in orange juice involves a multi-target action, which causes DNA damage, altered permeability and depolarization of membrane and compromised metabolic and efflux activities.

Control of Autochthonous Spoilage Lactic Acid Bacteria in Apple and Orange Juices by Sensorially Accepted Doses of Citrus Spp. Essential Oils Combined with Mild Heat Treatments.

This study assessed the compromised acceptance threshold (CAT) and rejection threshold (RT) of Citrus lemon (CLEO) and Citrus reticulata essential oil (CREO) in apple and orange juices. The efficacy of CLEO and CREO concentrations below the RT were evaluated alone and combined with mild heat treatment (MHT) (54 °C, up to 12 min) to inactivate the autochthonous spoilage bacteria Lactobacillus brevis, Lactobacillus plantarum, and Leuconostoc mesenteroides in apple and orange juices. The CAT of CLEO and CREO varied from 0.15 to 0.17 µL/mL in orange and apple juices. The RT of CLEO was approximately 0.58 µL/mL in apple and orange juices, and the RT of CREO was 0.68 µL/mL in both juices. When CLEO and CREO were assayed alone, the highest concentration (0.50 µL/mL) decreased counts of all strains approximately 2 log10 CFU/mL after 12 min of exposure to 54 °C. All concentrations of CLEO or CREO in combination with MHT acted synergistically against L. brevis, L. plantarum, and L. mesenteroides. Decreases in counts varied with the strain, CLEO and CREO concentrations, juice type, and exposure time to the combined treatment. CREO was more effective than CLEO in combination with MHT against the strains in apple and orange juices. Effective combinations of CLEO or CREO with MHT to control the autochthonous spoilage bacteria did not compromise the quality parameters (°Brix, pH, and titratable acidity) that characterize unsweetened juices. These results indicate CLEO or CREO at concentrations below the sensory RT in combination with MHT as a feasible technology to control autochthonous spoilage bacteria in fresh fruit juices. PRACTICAL APPLICATION: The present study provides novel information concerning the efficacy of sensorially accepted doses of CLEO and CREO combined with MHT against autochthonous spoilers in fruit juice. The valuable synergistic effects that can be observed when combining CLEO and CREO with MHT reveal a feasible preservation technology and alternative to traditional treatments that are successful because they help reduce treatment intensity, thereby avoiding adverse effects on the sensory, physicochemical, and nutritional properties of these products.