Pathogenic potential of the surviving Salmonella Enteritidis on strawberries after disinfection treatments based on ultraviolet-C light and peracetic acid.

Fresh fruits, especially strawberries, are usually consumed raw without any step to ensure their food safety. Salmonella enterica is one of the most important etiologic agents for foodborne diseases throughout the world and its ability to respond to some stress responses makes it even more dangerous. In the present investigation, we study the survival of S. Enteritidis (CECT-4300) on strawberries after 2-min of various disinfection steps (NaClO (200 ppm), peracetic acid (PAA; 40 ppm), water-assisted UV-C (WUV-C), and the combination WUV-C and 40 ppm of PAA (WUV-C + PAA)) and after 5 days of cold storage (4 °C). Moreover, the pathogenic potential of the surviving bacteria, such as the ability to survive throughout the gastrointestinal tract (GI) and subsequently the capability to adhere to and invade Caco-2 cells, was tested at each sampling point. After 2-min of washing procedures, reductions of S. Enteritidis on strawberries were ≥1.2 log, with no significant differences among treatments. However, the use of WUV-C + PAA treatment achieved the highest reductions in washing water, in which S. Enteritidis was not detected (

Evaluation of a sanitizing washing step with different chemical disinfectants for the strawberry processing industry.

Strawberries are often consumed fresh or only receive minimal processing, inducing a significant health risk to the consumer if contamination occurs anywhere from farm to fork. Outbreaks of foodborne illness associated with strawberries often involve a broad range of microbiological agents, from viruses (human norovirus) to bacteria (Salmonella spp. and Listeria monocytogenes). The addition of sanitizers to water washes is one of the most commonly studied strategies to remove or inactivate pathogens on berries as well as avoid cross contamination due to reuse of process wash water. The risk posed with the safety issues of by-products from chlorine disinfection in the fruit industry has led to a search for alternative sanitizers. We evaluated the applicability of different chemical sanitizers (peracetic acid (PA), hydrogen peroxide (H2O2), citric acid (CA), lactic acid (LA) and acetic acid (AA)) for the inactivation of S. enterica, L. monocytogenes and murine norovirus (MNV-1) on strawberries. A control treatment with chlorine (NaClO) (100 ppm) was included. For each sanitizer, different doses (40, 80 and 120 ppm for PA and 1, 2.5 and 5% for H2O2, LA, AA and CA) and time (2 and 5 min) were studied in order to optimize the decontamination washing step. The best concentrations were 80 ppm for PA, 5% for H2O2 and 2.5% for organic acids (LA, AA and CA) after 2 min treatment. Results indicate that the sanitizers selected may be a feasible alternative to chlorine (100 ppm) for removing selected pathogenic microorganisms (P > 0.05), with reductions about ≥2 log for bacterial strains and ≥ 1.7 log for MNV-1. As the washing water may also increase the microbial counts by cross-contamination, we observed that no pathogenic bacteria were found in wash water after 5% H2O2 and 80 ppm PA after 2 min treatment. On the other hand, we also reported reductions about total aerobic mesophyll (TAM) (0.0-1.4 log CFU/g) and molds and yeasts (M&Y) (0.3-1.8 log CFU/g) with all alternative sanitizers tested. Strawberries treated did not shown significant differences about physio-chemical parameters compared to the untreated samples (initial). For this study, the optimal sanitizer selected was PA, due to the low concentration and cost needed and its microbiocidal effect in wash water and fruit. Notwithstanding the results obtained, the effect of PA in combination with other non-thermal technologies such as water-assisted ultraviolet (UV-C) light should be studied in future research to improve the disinfection of strawberries.

Microbial interaction between Salmonella enterica and main postharvest fungal pathogens on strawberry fruit.

The microbial interaction between Salmonella enterica and the main postharvest fungal pathogens of strawberries was evaluated. Inoculation of fungal suspension was done 2 (D2) and 1 (D1) day(s) before and at the same time (D0) as S. enterica. Fruits were stored at 20 °C and 4 °C. At both temperatures, Botrytis cinerea and Rhizopus stolonifer caused a decrease in S. enterica population. Treatments where the mould was inoculated (D2, D1 and D0) achieved a significant logarithmic reduction (P < 0.05) of S. enterica populations after 48 h (20 °C) and 14 days (4 °C) compared to fungal-uninoculated fruits (CK). Regarding temperature, average reductions were significantly higher at 4 °C (3.38 log10 CFU/wound) than at 20 °C (1.16 log10 CFU/wound) (P < 0.05). Average reductions comprising all treatments were 1.91 and 0.41 log10 CFU/wound for B. cinerea and R. stolonifer at 20 °C, and 3.39 and 3.37 log10 CFU/wound for B. cinerea and R. stolonifer at 4 °C. A linear log10 model was fitted in order to predict the inactivation rate (kmax, log10 CFU/h) of S. enterica. Inactivation rates were higher at 20 °C for D2 treatments than at 4 °C throughout the running time. The main inactivation rate was obtained for B. cinerea at 20 °C (0.160 ±â€¯0.027/h), which was found to have stronger inhibitory activity against S. enterica than R. stolonifer. Univariate analysis ANOVA was carried out to evaluate the effect of different external variables on the inhibition of S. enterica. Results found that single effects were significant (P < 0.05) except for the pH. The inhibitory effect caused by the action of moulds in conjunction with some environmental factors could indicate the potential interactions between strawberry fungal pathogens and S. enterica.

Occurrence of selected viral and bacterial pathogens and microbiological quality of fresh and frozen strawberries sold in Spain.

Strawberry production and exports have been increasing in Spain in recent decades. However, little information is available about their microbiological quality. Due to the growing concern about the microbial safety of these fruits, the objective of this investigation was to study the microbiological quality and the prevalence of the main foodborne pathogens on strawberries sold in Spain. Fresh (n = 152) and frozen (n = 31) samples were obtained from marketplaces and fields in 2017 and 2018. The samples were assayed for total aerobic mesophilic microorganisms (TAM), moulds and yeasts (M&Y), total coliforms (TC), Escherichia coli, Salmonella spp., Listeria monocytogenes as well as Norovirus (NoV) GI and GII. The microbiological counts ranged from <1.70 (detection limit, dl) - 5.89 log10 CFU/g (mean 3.78 log10 CFU/g) for TAM; 2.10-5.86 log10 CFU/g (mean 3.80 log10 CFU/g) for M&Y; and <0.70 (dl) - 4.91 log10 CFU/g (mean 2.15 log10 CFU/g) for TC in fresh strawberries. In frozen strawberries, the counts were <1.70 (dl) - 3.66 log10 CFU/g (mean 2.30 log10 CFU/g) for TAM; <1.70 (dl) - 2.76 log10 CFU/g (mean 1.82 log10 CFU/g) for M&Y; and <0.70(dl) - 1.74 log10 CFU/g (mean 0.77 log10 CFU/g) for TC. All the samples in this study tested negative for Salmonella spp., L. monocytogenes. E. coli and NoV GI and GII genome. A global overview of all the data was executed using Principal Component Analysis (PCA), and the results showed that the scores and loadings according to principal components 1 (PC1) and 2 (PC2) accounted for 75.9% of the total variance, allowing a distinction between fresh and frozen samples. The presence of moulds was significantly higher in the supermarket samples whereas the presence of total coliforms was significantly higher in the field samples (p < 0.05). Although pathogenic microorganisms were not found, preventative measures and prerequisites in the strawberry production chain must be considered in order to avoid possible foodborne diseases related to the microbiological quality of the fruit.

Biopreservative methods to control the growth of foodborne pathogens on fresh-cut lettuce.

Fruits and vegetables can become contaminated by foodborne pathogens such as Escherichia coli O157:H7, Salmonella and Listeria monocytogenes, and it has been demonstrated that current industrial sanitizing treatments do not eliminate the pathogens when present. Chemical control is widely used, but biological control appears to be a better solution, mainly using the native microbiota present on fresh produce. The first objective of this study was to isolate native microbiota from whole and fresh-cut produce and to determine whether these bacteria were antagonistic toward foodborne pathogens. A total of 112 putative antagonist isolates were screened for their ability to inhibit the growth of Salmonella enterica on lettuce disks. Five different genera reduced S. enterica growth more than 1-log unit at 20°C at the end of 3 days. When tested against L. monocytogenes 230/3, only Pseudomonas sp. strain M309 (M309) was able to reduce pathogen counts by more than 1-log unit. Therefore, M309 strain was selected to be tested on lettuce disks at 10°C against S. enterica, E. coli O157:H7 and L. monocytogenes. M309 strain was only able to reduce S. enterica and E. coli O157:H7 populations. The second objective was to test different biopreservative methods including M309 strain, Pseudomonas graminis CPA-7 (CPA-7), bacteriophages (Listex P100 and Salmonelex) and nisin at conditions simulating commercial applications against Salmonella and L. monocytogenes on fresh-cut lettuce. The addition of the biopreservative agents did not result in a significant reduction of Salmonella population. However, CPA-7 strain together with nisin reduced L. monocytogenes numbers after 6 days of storage at 10°C. The cocktail of Salmonella and L. monocytogenes was not markedly inactivated by their respective bacteriophage solutions. This study highlighted the potential of biocontrol, but the combination with other technologies may be required to improve their application on fresh-cut lettuce.