The influence of pH on the efficacy of oxidation-reduction potential (ORP) to predict chlorine disinfection of surrogate bacteria, Escherichia coli O157:H7 and Listeria monocytogenes in oxidant demand free conditions and fresh produce wash water.

Oxidation-reduction potential (ORP) is commonly used as a rapid measurement of the antimicrobial potential of free chlorine during industrial fresh produce washing. The current study tested the hypothesis that ORP can act as a "single variable" measurement of bacterial (vegetative and endospores) inactivation effectiveness with free chlorine irrespective of the water pH value. This situation has on occasion been assumed but never confirmed nor disproven. Chlorine-dosed pH 6.5 and 8.5 phosphate buffer solutions were inoculated with Escherichia coli (E. coli), Listeria innocua (L. innocua), or Bacillus subtilis (B. subtilis) endospores. ORP, free chlorine (FC), and log reduction were monitored after 5 s (for E. coli and L. innocua) and up to 30 min (for B. subtilis spores) of disinfection. Logistic and exponential models were developed to describe how bacteria reduction varied as a function of ORP at different pH levels. Validation tests were performed in phosphate buffered pH 6.5 and 8.5 cabbage wash water periodically dosed with FC, cabbage extract and a cocktail of Escherichia coli O157:H7 (E. coli O157:H7) and Listeria monocytogenes (L. monocytogenes). The built logistic and exponential models confirmed that at equal ORP values, the inactivation of the surrogate strains was not consistent across pH 6.5 and pH 8.5, with higher reductions at higher pH. This is the opposite of the well-known free chlorine-controlled bacterial inactivation, where the antibacterial effect is higher at lower pH. The validation test results indicated that in the cabbage wash water, the relationship between disinfection efficiency and ORP was consistent with the oxidant demand free systems. The study suggests that ORP cannot serve as a reliable single variable measurement to predict bacterial disinfection in buffered systems. When using ORP to monitor and control the antibacterial effectiveness of the chlorinated wash water, it is crucial to take into account (and control) the pH.

Datasets for the environmental assessment of an apple value chain including fresh fruits, juice and applesauce from an organic low-input production farm.

Due to societal concerns, assess the environmental impacts, address the issues and provide labelling to the consumer are growing issues for the agri-food sector. In this context, provide datasets specific to alternative systems is crucial to be able to take into account the variability between systems then address their issues and label them appropriately. This data paper compiles all the data used to produce the life cycle assessment (LCA) environmental of an organic low-input apple value chain including the cultivation of apples at farm, the transformation of a part into juice and applesauce, the retail and the consumption stages. The raw data have mostly been obtained through interviews of the farmer and complemented by literature. They have been used to build a life cycle inventory (LCI), using Agribalyse 3.0 and Ecoinvent 3.8 as background databases. The dataset also compiles the life cycle impact assessment (LCIA) using the characterization method EF3.0. As discussed in an associated scientific paper, this dataset participates in filling two gaps: integrate the variability between systems in the discussion and link upstream (at farm) and downstream (transformation, retail, consuming) impacts. This is done by (1) covering the entire value chain from cradle to grave when most papers found in literature focusses on one stage (e.g. the cultivation of apples) and (2) applying LCA to a system that present specificities not well covered by LCA literature (e.g. low-input cultivation with no fertilization up to now).

Effect of ultrasonic treatment on enzymes: Decoupling the relation between the ultrasonic driven conformational change and enzyme activity.

Considering the growing interest in enzyme-based extraction technology as a safe and eco-friendly extraction technique, along with the relatively high cost associated with enzymatic applications, it became necessary to explore novel strategies aimed to improve enzyme activity. In this study, the impact of ultrasonic treatment on commercial cellulase and pectinase was investigated. As this effect may be influenced by various ultrasonic and enzyme-related parameters, changes in enzyme conformation were explored under optimal and non-optimal enzyme conditions. The intrinsic fluorescence spectrum was utilized as a tool for monitoring these changes. Additionally, the enzyme's catalytic potential was also assessed under the same conditions. Results indicated that the impact of ultrasonic treatment on enzyme conformation primarily depends on the total ultrasonic energy delivered to the system, rather than other ultrasonic parameters such as power, sample volume, treatment time, or duty cycle. The maximum relative decrease in intrinsic fluorescence intensity of Pectinex® Ultra Clear (PUC) and Pectinex® Ultra SPL (PUS) after ultrasonic treatment was approximately 51% and 55%, respectively, while the decrease induced by thermal denaturation was 25% and 30% respectively. Furthermore, a blue shift in the fluorescence spectrum of both pectinases was observed upon sonication for all process conditions indicating a change in enzyme conformation. However, ultrasonic treatment did not result in a significant change in enzyme activity, suggesting that these conformational adjustments may occur in regions other than the active sites. Moreover, ultrasonicated pectinases and cellulases did not exhibit any improvement in their catalytic potential under either optimal or non-optimal conditions.

Listeria monocytogenes in food businesses: From persistence strategies to intervention/prevention strategies-A review.

In 2023, Listeria monocytogenes persistence remains a problem in the food business. A profound understanding of how this pathogen persists may lead to better aimed intervention/prevention strategies. The lack of a uniform definition of persistence makes the comparison between studies complex. Harborage sites offer protection against adverse environmental conditions and form the ideal habitat for the formation of biofilms, one of the major persistence strategies. A retarded growth rate, disinfectant resistance/tolerance, desiccation resistance/tolerance, and protozoan protection complete the list of persistence strategies for Listeria monocytogenes and can occur on themselves or in combination with biofilms. Based on the discussed persistence strategies, intervention strategies are proposed. By enhancing the focus on four precaution principles (cleaning and disinfection, infrastructure/hygienic design, technical maintenance, and work methodology) as mentioned in Regulation (EC) No. 852/2004, the risk of persistence can be decreased. All of the intervention strategies result in obtaining and maintaining a good general hygiene status throughout the establishment at all levels ranging from separate equipment to the entire building.

Pesticide residues in (treated) wastewater and products of Belgian vegetable- and potato processing companies.

Pesticides are broadly utilized in crop cultivation and could end up in wastewater of vegetable- and potato companies during water-consuming processing steps. To gain insight into the presence of pesticide residues in (waste)water of these industries, water was analysed and monitored from three vegetable- and two potato processing companies in Belgium. During one year samples were collected of water before and after primary/secondary treatment (i.e. influent and effluent) and after tertiary treatment. Next to water, also (processed) carrot and potato products were analysed. Results show that boscalid (maximum: 18.32 µg/L) and terbuthylazine (maximum: 87.99 µg/L) are predominantly present in the vegetable industry and chlorpropham (maximum: 8.80×106 µg/L) and terbuthylazine (maximum: 3.37×105 µg/L) in the potato industry. The conventional treatment techniques seem to be insufficient for the removal of pesticides. Concentrations were even higher in the effluent than in the influent. Also, tertiary treatment techniques as ultra-filtration and reverse osmosis fail to reduce all pesticides below the European potable water limit of 0.1 µg/L. To meet this standard, the development and validation of new removal techniques are essential. Regarding product samples, almost no pesticide residues exceeded the MRL. Chlorpropham concentrations were statistically confirmed to be higher in potatoes and wastewater sampled when stored potatoes are processed.

Encapsulation of Lactobacillus in Low-Methoxyl Pectin-Based Microcapsules Stimulates Biofilm Formation: Enhanced Resistances to Heat Shock and Simulated Gastrointestinal Digestion.

Encapsulation is a common approach to improve the bacterial survival of probiotics. In this study, two new low-methoxyl pectins (CMP-6 and CMP-8) were used as coating materials to produce microcapsules (MCs) for the encapsulation of Lactobacillus acidophilus LMG9433T, Lactobacillus casei LMG6904T, and Lactobacillus rhamnosus LMG25859. A fermentation test showed that encapsulation did not influence the fermentation ability of lactobacilli. The biofilm formation of encapsulated lactobacilli was stimulated when an in situ cultivation was conducted on MCs, which was verified by cryo-SEM observation. The resultant biofilm-forming MCs (BMCs) contained high-density bacterial cells (∼1010 CFU/mL). Compared to planktonic lactobacilli, pectin-based MCs showed significant protection for encapsulated lactobacilli from heat shock and simulated gastric digestion. Especially, benefiting from the biofilm formation, BMCs provided higher protection with enhanced resistance to heat shock, freeze-drying, and gastrointestinal digestion than MCs. Our result highlighted the superior bacterial resistances of biofilm-forming probiotics encapsulated in pectinate microcapsules.

The public health risk posed by Listeria monocytogenes in frozen fruit and vegetables including herbs, blanched during processing.

A multi-country outbreak of Listeria monocytogenes ST6 linked to blanched frozen vegetables (bfV) took place in the EU (2015-2018). Evidence of food-borne outbreaks shows that L. monocytogenes is the most relevant pathogen associated with bfV. The probability of illness per serving of uncooked bfV, for the elderly (65-74 years old) population, is up to 3,600 times greater than cooked bfV and very likely lower than any of the evaluated ready-to-eat food categories. The main factors affecting contamination and growth of L. monocytogenes in bfV during processing are the hygiene of the raw materials and process water; the hygienic conditions of the food processing environment (FPE); and the time/Temperature (t/T) combinations used for storage and processing (e.g. blanching, cooling). Relevant factors after processing are the intrinsic characteristics of the bfV, the t/T combinations used for thawing and storage and subsequent cooking conditions, unless eaten uncooked. Analysis of the possible control options suggests that application of a complete HACCP plan is either not possible or would not further enhance food safety. Instead, specific prerequisite programmes (PRP) and operational PRP activities should be applied such as cleaning and disinfection of the FPE, water control, t/T control and product information and consumer awareness. The occurrence of low levels of L. monocytogenes at the end of the production process (e.g. < 10 CFU/g) would be compatible with the limit of 100 CFU/g at the moment of consumption if any labelling recommendations are strictly followed (i.e. 24 h at 5°C). Under reasonably foreseeable conditions of use (i.e. 48 h at 12°C), L. monocytogenes levels need to be considerably lower (not detected in 25 g). Routine monitoring programmes for L. monocytogenes should be designed following a risk-based approach and regularly revised based on trend analysis, being FPE monitoring a key activity in the frozen vegetable industry.

Effect of mild heat treatment on browning-related parameters in fresh-cut Iceberg lettuce.

Enzymatic browning of Iceberg lettuce was studied by subjecting midrib tissues to a series of mild heat treatments. The effects of wounding and subsequent application of a mild heat treatment were examined by monitoring the browning potential (BP) and the activity of three browning-related enzymes (i.e., phenylalanine ammonia lyase [PAL], polyphenol oxidase [PPO], and peroxidase [POD]) during refrigerated storage up to 10 days. Efficient inhibition of browning was achieved by treatment at 50°C for 60 s. The wound-induced increase of the BP and the activity of PAL and POD was effectively suppressed, maintaining their values at initial levels up to 7 days of storage. PPO activity, on the contrary, remained unchanged after wounding, whether or not followed by heat treatment. BP, PAL activity and POD were found to be strongly correlated, whereas meaningful associations for PPO with the other parameters could not be established. PRACTICAL APPLICATIONS: In an attempt to answer to the growing demand in the fresh-cut produce industry to control browning, heat treatment was investigated as interesting alternative to chemical preservation methods. Efficient control of enzymatic browning in fresh-cut Iceberg lettuce could be achieved by heat treatment at 50°C for 60 s. Experimental data are provided showing the effects of wounding and subsequent heat treatment on visual browning, the BP and the activity of PAL, PPO, and POD during refrigerated storage up to 10 days. Using this data, correlations were found for BP, PAL activity, and POD activity, but not for PPO. Although undesired side effects of heat treatment (e.g., tissue softening) cannot be excluded, the obtained information might be useful for further research, serving as a baseline for wound-induced effects on browning-related parameters in fresh-cut lettuce and possible mechanisms of action of inhibitory treatments.

Occurrence and characterisation of biofilms in drinking water systems of broiler houses.

BACKGROUND: Water quality in the drinking water system (DWS) plays an important role in the general health and performance of broiler chickens. Conditions in the DWS of broilers are ideal for microbial biofilm formation. Since pathogens might reside within these biofilms, they serve as potential source of waterborne transmission of pathogens to livestock and humans. Knowledge about the presence, importance and composition of biofilms in the DWS of broilers is largely missing. In this study, we therefore aim to monitor the occurrence, and chemically and microbiologically characterise biofilms in the DWS of five broiler farms. RESULTS: The bacterial load after disinfection in DWSs was assessed by sampling with a flocked swab followed by enumerations of total aerobic flora (TAC) and Pseudomonas spp. The dominant flora was identified and their biofilm-forming capacity was evaluated. Also, proteins, carbohydrates and uronic acids were quantified to analyse the presence of extracellular polymeric substances of biofilms. Despite disinfection of the water and the DWS, average TAC was 6.03 ± 1.53 log CFU/20cm2. Enumerations for Pseudomonas spp. were on average 0.88 log CFU/20cm2 lower. The most identified dominant species from TAC were Stenotrophomonas maltophilia, Pseudomonas geniculata and Pseudomonas aeruginosa. However at species level, most of the identified microorganisms were farm specific. Almost all the isolates belonging to the three most abundant species were strong biofilm producers. Overall, 92% of all tested microorganisms were able to form biofilm under lab conditions. Furthermore, 63% of the DWS surfaces appeared to be contaminated with microorganisms combined with at least one of the analysed chemical components, which is indicative for the presence of biofilm. CONCLUSIONS: Stenotrophomonas maltophilia, Pseudomonas geniculata and Pseudomonas aeruginosa are considered as opportunistic pathogens and could consequently be a potential risk for animal health. Additionally, the biofilm-forming capacity of these organisms could promote attachment of other pathogens such as Campylobacter spp. and Salmonella spp.

A review on influencing factors on the minimum inhibitory concentration of essential oils.

With growing interest in essential oils as natural preservatives in the food industry, the literature is expanding enormously. To understand the antimicrobial activity of essential oils, the antimicrobial mechanism of individual essential oil (EO) compounds, and their minimum inhibitory concentrations (MICs), are interesting starting points for research. Therefore, and to get insight into the factors influencing their antimicrobial activities, the Web of Science was searched for MICs of EO compounds (1995-2016). Many MICs for individual EO compounds have already been reported in the literature, but there is large variability in these data, even for the MIC of the same compound against the same species. No correlation was found between the tested structural parameters of EO compounds (polarity, water solubility, dissociation constant, molecular weight and molecular complexity) and their MICs against all microorganisms, Gram-negative bacteria, Gram-positive bacteria and fungi. Few clear differences in sensitivity between microorganisms could be found. Based on this review it is clear that different incubation conditions, culture media and the use of emulsifiers/solvents have an influence on the MIC, causing big variance. This review points out the need for a good international standard method to assess the antimicrobial activity of EO compounds for better comparability between studies.

Evaluation of Two Surface Sampling Methods for Microbiological and Chemical Analyses To Assess the Presence of Biofilms in Food Companies.

Biofilms are an important source of contamination in food companies, yet the composition of biofilms in practice is still mostly unknown. The chemical and microbiological characterization of surface samples taken after cleaning and disinfection is very important to distinguish free-living bacteria from the attached bacteria in biofilms. In this study, sampling methods that are potentially useful for both chemical and microbiological analyses of surface samples were evaluated. In the manufacturing facilities of eight Belgian food companies, surfaces were sampled after cleaning and disinfection using two sampling methods: the scraper-flocked swab method and the sponge stick method. Microbiological and chemical analyses were performed on these samples to evaluate the suitability of the sampling methods for the quantification of extracellular polymeric substance components and microorganisms originating from biofilms in these facilities. The scraper-flocked swab method was most suitable for chemical analyses of the samples because the material in these swabs did not interfere with determination of the chemical components. For microbiological enumerations, the sponge stick method was slightly but not significantly more effective than the scraper-flocked swab method. In all but one of the facilities, at least 20% of the sampled surfaces had more than 102 CFU/100 cm2. Proteins were found in 20% of the chemically analyzed surface samples, and carbohydrates and uronic acids were found in 15 and 8% of the samples, respectively. When chemical and microbiological results were combined, 17% of the sampled surfaces were contaminated with both microorganisms and at least one of the analyzed chemical components; thus, these surfaces were characterized as carrying biofilm. Overall, microbiological contamination in the food industry is highly variable by food sector and even within a facility at various sampling points and sampling times.

Risk Factors for Salmonella, Shiga Toxin-Producing Escherichia coli and Campylobacter Occurrence in Primary Production of Leafy Greens and Strawberries.

The microbiological sanitary quality and safety of leafy greens and strawberries were assessed in the primary production in Belgium, Brazil, Egypt, Norway and Spain by enumeration of Escherichia coli and detection of Salmonella, Shiga toxin-producing E. coli (STEC) and Campylobacter. Water samples were more prone to containing pathogens (54 positives out of 950 analyses) than soil (16/1186) and produce on the field (18/977 for leafy greens and 5/402 for strawberries). The prevalence of pathogens also varied markedly according to the sampling region. Flooding of fields increased the risk considerably, with odds ratio (OR) 10.9 for Salmonella and 7.0 for STEC. A significant association between elevated numbers of generic E. coli and detection of pathogens (OR of 2.3 for STEC and 2.7 for Salmonella) was established. Generic E. coli was found to be a suitable index organism for Salmonella and STEC, but to a lesser extent for Campylobacter. Guidelines on frequency of sampling and threshold values for E. coli in irrigation water may differ from region to region.

Effect of Disinfectants on Preventing the Cross-Contamination of Pathogens in Fresh Produce Washing Water.

The potential cross-contamination of pathogens between clean and contaminated produce in the washing tank is highly dependent on the water quality. Process wash water disinfectants are applied to maintain the water quality during processing. The review examines the efficacy of process wash water disinfectants during produce processing with the aim to prevent cross-contamination of pathogens. Process wash water disinfection requires short contact times so microorganisms are rapidly inactivated. Free chlorine, chlorine dioxide, ozone, and peracetic acid were considered suitable disinfectants. A disinfectant's reactivity with the organic matter will determine the disinfectant residual, which is of paramount importance for microbial inactivation and should be monitored in situ. Furthermore, the chemical and worker safety, and the legislative framework will determine the suitability of a disinfection technique. Current research often focuses on produce decontamination and to a lesser extent on preventing cross-contamination. Further research on a sanitizer's efficacy in the washing water is recommended at the laboratory scale, in particular with experimental designs reflecting industrial conditions. Validation on the industrial scale is warranted to better understand the overall effects of a sanitizer.

Decontamination of Pangasius fish (Pangasius hypophthalmus) with chlorine or peracetic acid in the laboratory and in a Vietnamese processing company.

This study evaluated the decontamination of Pangasius fillets in chlorine or peracetic acid treated wash water. First, the decontamination efficacy of the washing step with chlorinated water applied by a Vietnamese processing company during trimming of Pangasius fillets was evaluated and used as the basis for the experiments performed on a laboratory scale. As chlorine was only added at the beginning of the batch and used continuously without renewal for 239min; a rapid increase of the bacterial counts and a fast decrease of chlorine in the wash water were found. This could be explained by the rapid accumulation of organic matter (ca. 400mg O2/L of COD after only 24min). Secondly, for the experiments performed on a laboratory scale, a single batch approach (one batch of wash water for treating a fillet) was used. Chlorine and PAA were evaluated at 10, 20, 50 and 150ppm at contact times of 10, 20 and 240s. Washing with chlorine and PAA wash water resulted in a reduction of Escherichia coli on Pangasius fish which ranged from 0-1.0 and 0.4-1.4logCFU/g, respectively while less to no reduction of total psychrotrophic counts, lactic acid bacteria and coliforms on Pangasius fish was observed. However, in comparison to PAA, chlorine was lost rapidly. As an example, 53-83% of chlorine and 15-17% of PAA were lost after washing for 40s (COD=238.2±66.3mg O2/L). Peracetic acid can therefore be an alternative sanitizer. However, its higher cost will have to be taken into consideration. Where (cheaper) chlorine is used, the processors have to pay close attention to the residual chlorine level, pH and COD level during treatment for optimal efficacy.

Selection criteria for water disinfection techniques in agricultural practices.

This paper comprises a selection tool for water disinfection methods for fresh produce pre- and postharvest practices. A variety of water disinfection technologies is available on the market and no single technology is the best choice for all applications. It can be difficult for end users to choose the technology that is best fit for a specific application. Therefore, the different technologies were characterized in order to identify criteria that influence the suitability of a technology for pre- or postharvest applications. Introduced criteria were divided into three principal components: (i) criteria related to the technology and which relate to the disinfection efficiency, (ii) attention points for the management and proper operation, and (iii) necessities in order to sustain the operation with respect to the environment. The selection criteria may help the end user of the water disinfection technology to obtain a systematic insight into all relevant aspects to be considered for preliminary decision making on which technologies should be put to feasibility testing for water disinfection in pre- and postharvest practices of the fresh produce chain.

Agricultural and management practices and bacterial contamination in greenhouse versus open field lettuce production.

The aim of this study was to gain insight into potential differences in risk factors for microbial contamination in greenhouse versus open field lettuce production. Information was collected on sources, testing, and monitoring and if applicable, treatment of irrigation and harvest rinsing water. These data were combined with results of analysis on the levels of Escherichia coli as a fecal indicator organism and the presence of enteric bacterial pathogens on both lettuce crops and environmental samples. Enterohemorragic Escherichia coli (EHEC) PCR signals (vt1 or vt2 positive and eae positive), Campylobacter spp., and Salmonella spp. isolates were more often obtained from irrigation water sampled from open field farms (21/45, 46.7%) versus from greenhouse production (9/75, 12.0%). The open field production was shown to be more prone to fecal contamination as the number of lettuce samples and irrigation water with elevated E. coli was significantly higher. Farmers comply with generic guidelines on good agricultural practices available at the national level, but monitoring of microbial quality, and if applicable appropriateness of water treatment, or water used for irrigation or at harvest is restricted. These results indicate the need for further elaboration of specific guidelines and control measures for leafy greens with regard to microbial hazards.

Relationships among hygiene indicators and enteric pathogens in irrigation water, soil and lettuce and the impact of climatic conditions on contamination in the lettuce primary production.

Eight Belgian lettuce farms located in the West Flanders were sampled to establish the relationships between levels of indicator bacteria, detection of enteric zoonotic pathogens and the temperature and precipitation during primary production. Pathogenic bacteria (PCR EHEC positives, Salmonella spp. or Campylobacter spp.) and indicator bacteria (total psychrotrophic aerobic plate count (TPAC), total coliforms, Escherichia coli, enterococci) were determined over a period of one and a half year from seedling leaves, peat-soil of the seedling, lettuce crops, field soil and irrigation water. Neither Salmonella isolates nor PCR EHEC signals were detected from lettuce although one out of 92 field soil samples contained Salmonella spp. and five soil samples provided PCR positives for EHEC virulence factors (vt1 or vt2 and eae gene). A low prevalence of Campylobacter (8/88) was noted in lettuce. It was shown that irrigation water is a major risk factor with regard to the bacterial contamination of the fresh produce as the water samples showed on a regular basis E. coli presence (59.2% of samples≥1CFU/100ml) and occasionally detection of pathogens (25%, n=30/120), in particular Campylobacter spp. The highest correlations between indicator bacteria, pathogens, temperature and the amount of precipitation were observed for the water samples in contrast to the soil or lettuce samples where no correlations were observed. The high correlations between E. coli, total coliforms and enterococci in the water implicated redundancy between analyses. Presence of elevated levels of E. coli increased the probability for the presence of pathogens (Campylobacter spp., EHEC and Salmonella spp.), but had a low to moderate predictive value on the actual presence of pathogens. The presence of pathogens and indicator bacteria in the water samples showed a seasonal effect as they tend to be more present during the months with higher temperature.

Organic acid based sanitizers and free chlorine to improve the microbial quality and shelf-life of sugar snaps.

A screening in a sugar snap packaging company showed a converged build-up of aerobic psychrotrophic plate count (APC) (ca. 6.5 log CFU/100mL), yeasts and molds (Y&M), and lactic acid bacteria (LAB) (both ca. 4.5 log CFU/100mL) in the wash water in the absence of water sanitizer, and a low build-up of chemical oxygen demand (30 ± 5 mg O2/L) and turbidity (5.2 ± 1.1 NTU). Decontamination experiments were performed in the lab with Purac FCC 80® (80% L(+) lactic acid), two other commercial water sanitizers based on organic acids (NATRApHASe-ABAV®, and NATRApHASe-FVS®) and chlorine to evaluate their performance in reduction of the sugar snap microbial load as well as their functionality as disinfectant of the wash water to avoid cross-contamination. An additional 1 log reduction of APC on the sugar snaps was achieved with lactic acid in the range 0.8 to 1.6%, ABAV 0.5%, and free chlorine 200mg/L when compared to a water wash, while no significant difference in the numbers of Y&M was obtained when washing in sanitizer compared to water. There was no significant influence of the studied concentration and contact time on decontamination efficiency. Treatment with lactic acid 0.8% resulted in a lower APC contamination on the sugar snaps than on the untreated and water washed samples for 10 days. Chlorine 200mg/L was the only treatment able to maintain the Y&M load lower than the untreated samples throughout the entire storage duration. The use of water sanitizers could not extend the sensorial shelf-life. Microbial loads were not indicative/predictive of visual microbial spoilage (shelf-life limiting factor), whereas maturity and amount of damage at the calyx end of the pods were. The APC wash water contamination (5.2 log CFU/100mL) was reduced significantly by chlorine 20 to 200mg/L (to 1.4 log CFU/100mL), ABAV 0.5 to 1.5% (to 2.7 log CFU/100mL), FVS 0.5% (to 2.7 log CFU/100mL) and lactic acid 0.8 to 1.6% (to 3.4 log CFU/100mL). Only the use of chlorine enabled the reduction of the Y&M wash water contamination significantly (from 3.4 to 1.4 log CFU/100mL). The low physicochemical build-up of the sugar snap wash water during the industrial washing process makes free chlorine attractive as a water disinfectant to prevent bacterial and fungal cross-contamination, whereas the sanitizers based on organic acids are not, due to their weak water disinfection efficiency.

Moderate prevalence of antimicrobial resistance in Escherichia coli isolates from lettuce, irrigation water, and soil.

Fresh produce is known to carry nonpathogenic epiphytic microorganisms. During agricultural production and harvesting, leafy greens can become contaminated with antibiotic-resistant pathogens or commensals from animal and human sources. As lettuce does not undergo any inactivation or preservation treatment during processing, consumers may be exposed directly to all of the (resistant) bacteria present. In this study, we investigated whether lettuce or its production environment (irrigation water, soil) is able to act as a vector or reservoir of antimicrobial-resistant Escherichia coli. Over a 1-year period, eight lettuce farms were visited multiple times and 738 samples, including lettuce seedlings (leaves and soil), soil, irrigation water, and lettuce leaves were collected. From these samples, 473 isolates of Escherichia coli were obtained and tested for resistance to 14 antimicrobials. Fifty-four isolates (11.4%) were resistant to one or more antimicrobials. The highest resistance rate was observed for ampicillin (7%), followed by cephalothin, amoxicillin-clavulanic acid, tetracycline, trimethoprim, and streptomycin, with resistance rates between 4.4 and 3.6%. No resistance to amikacin, ciprofloxacin, gentamicin, or kanamycin was observed. One isolate was resistant to cefotaxime. Among the multiresistant isolates (n = 37), ampicillin and cephalothin showed the highest resistance rates, at 76 and 52%, respectively. E. coli isolates from lettuce showed higher resistance rates than E. coli isolates obtained from soil or irrigation water samples. When the presence of resistance in E. coli isolates from lettuce production sites and their resistance patterns were compared with the profiles of animal-derived E. coli strains, they were found to be the most comparable with what is found in the cattle reservoir. This may suggest that cattle are a potential reservoir of antimicrobial-resistant E. coli strains in plant primary production.

Physicochemical quality and chemical safety of chlorine as a reconditioning agent and wash water disinfectant for fresh-cut lettuce washing.

Chlorine was assessed as a reconditioning agent and wash water disinfectant in the fresh-cut produce industry. Artificial fresh-cut lettuce wash water, made from butterhead lettuce, was used for the experiments. In the reconditioning experiments, chlorine was added to artificial wash water inoculated with Escherichia coli O157 (6 log CFU/ml). Regression models were constructed based on the inactivation data and validated in actual wash water from leafy vegetable processing companies. The model that incorporated chlorine dose and chemical oxygen demand (COD) of the wash water accurately predicted inactivation. Listeria monocytogenes was more resistant to chlorine reconditioning in artificial wash water than Salmonella spp. and Escherichia coli O157. During the washing process with inoculated lettuce (4 log CFU/g), in the absence of chlorine, there was a rapid microbial buildup in the water that accumulated to 5.4 ± 0.4 log CFU/100 ml after 1 h. When maintaining a residual concentration of 1 mg/liter free chlorine, wash water contamination was maintained below 2.7, 2.5, and 2.5 log CFU/100 ml for tap water and artificial process water with COD values of 500 and 1,000 mg O2/liter, respectively. A model was developed to predict water contamination during the dynamic washing process. Only minor amounts of total trihalomethanes were formed in the water during reconditioning. Total trihalomethanes accumulated to larger amounts in the water during the wash water disinfection experiments and reached 124.5 ± 13.4 µg/liter after 1 h of execution of the washing process in water with a COD of 1,000 mg O2/liter. However, no total trihalomethanes were found on the fresh-cut lettuce after rinsing.