The impact of extreme weather events on Salmonella internalization in lettuce and green onion

Salmonella internalization is an important issue in raw vegetable consumption because washing usually cannot remove or inactivate the internalized pathogens effectively. In this study, the impact of extreme weather events, drought and heavy rains, caused by climate change on the internalization of Salmonella Typhimurium was investigated. Two leafy green fresh produce, iceberg lettuce and green onion were chosen. Rhizosphere soil inoculation was conducted to mimic the contamination routes via soil and then root uptake. Most internalized S. Typhimurium were found in lettuce leaves and in the root portions of green onion under all three irrigation conditions (optimal, drought, storm). In general, high concentration of soil inoculation facilitated the internalization level in both lettuce and green onion. Under extreme weather conditions, the internalization of S. Typhimurium in lettuce occurred when the soil was contaminated with a high level of bacteria (8–9 log colony forming unit (CFU)/g soil) and under these conditions, the internalization level was higher than the lettuce grown at the optimal water condition, except with 8 log CFU/g contamination (storm). Under drought, the results showed high variation, but the level of internalization of S. Typhimurium in lettuce increased by 16 times (1.21 log CFU/g) and 27 times (1.43 log CFU/g) compared to the optimally irrigated group when the soil was contaminated with 8 log and 9 log CFU/g soil, respectively. Ten-fold increased internalization was observed in the over-irrigated lettuce leaves when the soil was contaminated with 9 log CFU/g soil. The green onion samples showed ~ 4 log CFU/g green onion of S. Typhimurium internalization when exposed to high level of contamination (> 7 log CFU/g soil), which is a much higher internalization rate than the lettuce (average 2–3 log CFU/g). However, from the green onion experiments, no apparent patterns of water stress that affect on the levels on the Salmonella internalization were observed.     

Understanding E. coli internalization in lettuce leaves for optimization of irradiation treatment

Irradiation penetrates food tissues and effectively reduces the number of food microorganisms in fresh produce, but the efficacy of the process against internalized bacteria is unknown. The objective of this study was to understand the mechanisms of pathogen colonization of plants relative to lettuce leaf structures so that radiation treatment of fresh leafy vegetables can be optimized. Leaves of iceberg, Boston, green leaf, and red leaf lettuces were cut into pieces, submerged in a cocktail mixture of two isolates of Escherichia coli (Rifampicin resistant), and subjected to a vacuum perfusion process to force the bacterial cells into the intercellular spaces in the leaves. Sixty bags containing 20 g of lettuce each were tested. The inoculated leaves were gamma irradiated (Lanthanum-140, 0.16 kGy/h) at 0.25–1.0-kGy (surface dose values), with increments of 0.25 kGy at 15 °C. Microbial analysis was performed right after irradiation, including non-irradiated leaf pieces (controls). A dose uniformity ratio (max/min dose) of 2.8 was set to confirm the effect of non-uniform dose distribution. Calculated D₁₀-values varied between 48 and 62% based on the dose distribution from the entrance dose. However, despite the subtle differences in composition and structure among the four lettuce varieties, the D₁₀-values were not significantly different. Irradiation up to 1.0-kGy resulted in 3–4-log reduction of internalized E. coli on the lettuce leaves. The SEM images suggest that the contamination sites of pathogens in leafy vegetables are mainly localized on crevices and into the stomata. This study shows that irradiation effectively reduces viable E. coli cells internalized in lettuce, and decontamination is not influenced by lettuce variety. Ionizing irradiation effectively reduced the population of internalized pathogen in a dose-dependent manner and could be used as an effective killing step to mitigate the risk of foodborne disease outbreaks.     

Salmonella surveillance and control at post-harvest in the Belgian pork meat chain

Salmonella remains the primary cause of reported bacterial food borne disease outbreaks in Belgium. Pork and pork products are recognized as one of the major sources of human salmonellosis. In contrast with the primary production and slaughterhouse phases of the pork meat production chain, only a few studies have focussed on the post-harvest stages. The goal of this study was to evaluate Salmonella and Escherichia coli contamination at the Belgian post-harvest stages. E. coli counts were estimated in order to evaluate the levels of faecal contamination. The results of bacteriological analysis from seven cutting plants, four meat-mincing plants and the four largest Belgian retailers were collected from official and self-monitoring controls. The prevalence of Salmonella in the cutting plants and meat-mincing plants ranged from 0% to 50%. The most frequently isolated serotype was Salmonella typhimurium. The prevalence in minced meat at retail level ranged from 0.3% to 4.3%. The levels of Salmonella contamination estimated from semi-quantitative analysis of data relating to carcasses, cuts of meat and minced meat were equal to −3.40 ± 2.04 log CFU/cm², −2.64 ± 1.76 log CFU/g and −2.35 ± 1.09 log CFU/g, respectively. The E. coli results in meat cuts and minced meat ranged from 0.21 ± 0.50 to 1.23 ± 0.89 log CFU/g and from 1.33 ± 0.58 to 2.78 ± 0.43 log CFU/g, respectively. The results showed that faecal contamination still needs to be reduced, especially in specific individual plants.     

Efficacy of adding detergents to sanitizer solutions for inactivation of Escherichia coli O157:H7 on Romaine lettuce

Numerous Escherichia coli O157:H7 outbreaks have been linked to consumption of fresh lettuce. The development of effective and easily implemented wash treatment could reduce such incidents. The purpose of this study was to evaluate the addition of food-grade detergents to sanitizer solutions for inactivation of E. coli O157:H7 on Romaine lettuce. Freshly-cut leaves of Romaine lettuce were dip-inoculated to achieve a final cell concentration of 7.8 ± 0.2 log CFU/g, air-dried for 2 h, and stored overnight at 4 °C. Leaves were then washed for 2 min in an experimental short chain fatty acid formulation (SCFA) or in one of the following solutions with or without 0.2% dodecylbenzenesulfonic acid or 0.2% sodium 2-ethyl hexyl sulfate: 1) deionized water; 2) 100 ppm chlorine dioxide; 3) 100 ppm chlorine; and 4) 200 ppm chlorine. Following wash treatment, samples were blended in neutralizing buffer (1:3) and surface plated on the selective media CT-SMAC. The efficacy of wash treatments, with or without the detergents, in inactivating E. coli O157:H7 cells on lettuce leaves were not significantly different. The most effective wash solution was SCFA, which was capable of reducing E. coli O157:H7 populations by more than 5 log CFU/g. The rest of the wash treatments resulted in a population reduction of less than 1 log CFU/g. The effectiveness of SCFA surpasses that of other sanitizer treatments tested in this study and requires further research to optimize treatments to preserve lettuce quality. Conventional detergents did not enhance the efficacy of any of the wash treatments tested during this study.     

Preharvest internalization of Escherichia coli O157:H7 into lettuce leaves, as affected by insect and physical damage

Environmental pests may serve as reservoirs and vectors of zoonotic pathogens to leafy greens; however, it is unknown whether insect pests feeding on plant tissues could redistribute these pathogens present on the surface of leaves to internal sites. This study sought to differentiate the degree of tissue internalization of Escherichia coli O157:H7 when applied at different populations on the surface of lettuce and spinach leaves, and to ascertain whether lettuce-infesting insects or physical injury could influence the fate of either surface or internalized populations of this enteric pathogen. No internalization of E. coli O157:H7 occurred when lettuce leaves were inoculated with 4.4 log CFU per leaf, but it did occur when inoculated with 6.4 log CFU per leaf. Internalization was statistically greater when spinach leaves were inoculated on the abaxial (underside) than when inoculated on the adaxial (topside) side, and when the enteric pathogen was spread after surface inoculation. Brief exposure (～18 h) of lettuce leaves to insects (5 cabbage loopers, 10 thrips, or 10 aphids) prior to inoculation with E. coli O157:H7 resulted in significantly reduced internalized populations of the pathogen within these leaves after approximately 2 weeks, as compared with leaves not exposed to insects. Surface-contaminated leaves physically injured through file abrasions also had significantly reduced populations of both total and internalized E. coli O157:H7 as compared with nonabraded leaves 2 weeks after pathogen exposure.     

Simple quantitative analysis of Escherichia coli K-12 internalized in baby spinach using Fourier Transform Infrared spectroscopy

Bacterial contamination continues to be a serious concern for food safety. Although washing fresh produce helps in reducing pathogen levels, pathogen internalization often limits the effectiveness of washing. When pathogens internalize in leafy vegetables, the method of identification and quantitative measurement would be called into question. This study was aimed to use Fourier Transform Infrared (FTIR) spectroscopy integrated with an attenuated total reflectance kit for quantification of Escherichia coli K-12 internalized in baby spinach. The bacteria were inoculated into vascular and intracellar tissues of spinach leaves by syringe injection and the distribution of internalized E. coli K-12 cells was confirmed under scanning electron microscopy (SEM). FTIR measurement following the preparation of bacterial suspension from spinach leaves with high speed pulverizing enabled to detect the absorbance peaks in the amide II region between 1590 and 1490 cm⁻¹ as a fingerprint for the microbes. It was found that the estimated concentrations of E. coli K-12 agreed well with the concentrations determined by plate counting with R² values of 0.98 and 0.97 in peptone water and spinach extracts, respectively. The results demonstrated that FTIR can identify and quantify E. coli K-12 in baby spinach extracts at a limit of detection of approximately 100 CFU/mL in 5 min. The developed method is expected to be suitable for the analysis of pathogenic E. coli strains and other bacterial species in fresh vegetables.     

Internalization of E. coli O157:H7 and Salmonella spp. in plants: A review

Extensive research efforts have been devoted in the last decade to providing insight into understanding the factors that have contributed to the increase in the number of outbreaks of Salmonella spp. and E. coli O157:H7 involving fresh produce, including leafy greens, fruits, and nuts. Many studies have shown that both Salmonella spp. and E. coli O157:H7 can internalize within a variety of tissue types and that there are numerous factors that can influence the extent of internalization. In this review, a compilation of the studies that have been performed to examine internalization, as well as various techniques that have been developed to identify the plant organs and tissues that harbor internalized bacteria are presented. In addition, the various routes of internalization into the plant, survival following internalization, and interactions between internalized bacteria and the plant as well as associated microbes are also addressed.     

Effects of X-ray radiation on Escherichia coli O157:H7, Listeria monocytogenes, Salmonella enterica and Shigella flexneri inoculated on shredded iceberg lettuce

The main goal of this investigation was to study the efficacy of X-ray doses (0.1, 0.2, 0.3, 0.5, 0.75, 1.0, 1.5 and 2.0 kGy) on inoculated Escherichia coli O157: H7, Listeria monocytogenes, Salmonella enterica and Shigella flexneri on shredded iceberg lettuce. The second goal was to study the effect of X-ray on the inherent microflora counts and visual color of shredded iceberg lettuce during storage at 4 °C for 30 days. Treatment with 1.0 kGy X-ray significantly reduced the population of E. coli O157: H7, L. monocytogenes, Salmonella enterica and S. flexneri on shredded iceberg lettuce by 4.4, 4.1, 4.8 and 4.4-log CFU 5 cm⁻², respectively. Furthermore, more than a 5 log CFU reduction of E. coli O157: H7, L. monocytogenes, S. enterica and S. flexneri was achieved with 2.0 kGy X-ray. Treatment with X-ray reduced the initial microflora on iceberg lettuce and kept them significantly (p < 0.05) lower than the control during storage at 4 °C and 90% RH for 30 days. Treatment with X-ray did not significantly (p > 0.05) change the green color of iceberg lettuce leaves. Treatment with X-ray significantly reduced selected pathogens and inherent microorganisms on shredded iceberg lettuce leaves, which could be a good alternative to other technologies for produce (lettuce) industry.     

Combined effect of ultrasound and organic acids to reduce Escherichia coli O157:H7, Salmonella Typhimurium, and Listeria monocytogenes on organic fresh lettuce

This study was performed to compare the effectiveness of individual treatments (ultrasound and organic acids) and their combination on reducing foodborne pathogens on organic fresh lettuce. Lettuce leaves were inoculated with a cocktail of three strains each of Escherichia coli O157:H7, Salmonella Typhimurium, and Listeria monocytogenes and treated with ultrasound (40 kHz) alone, organic acids (0.3, 0.5, 0.7, 1.0, and 2.0% — malic acid, lactic acid, and citric acid) alone and combined with ultrasound and organic acids for 5 min. For all 3 pathogens, the combined treatment of ultrasound and organic acids resulted in additional 0.8 to 1.0 log reduction compared to individual treatments, without causing significant quality change (color and texture) on lettuce during 7 day storage. The maximum reductions of E. coli O157:H7, S. Typhimurium, and L. monocytogenes were 2.75, 3.18, and 2.87 log CFU/g observed after combined treatment with ultrasound and 2% organic acid for 5 min, respectively. Our results suggest that the combined treatment of ultrasound with organic acids was effective at increasing pathogen reduction compared to individual treatments without significantly affecting quality, and demonstrates its potential as a novel method to increase the microbial safety on organic fresh lettuce.     

Composition and availability of soluble and insoluble oxalates in raw and cooked taro (Colocasia esculenta var. Schott) leaves

Taro (Colocasia esculenta var. Schott) is a major staple food crop in parts of Asia and the Pacific Islands and is grown as a minor crop in New Zealand. Soluble, insoluble and total oxalate content of young and older leaves were determined by HPLC following hot water (80 °C) and hot (80 °C) acid (0.2 mol/L HCL) extractions. Young taro leaves contained 589 ± 35.8 mg total oxalates/100 g fresh weight (FW) while older taro leaves contained (443 ± 15.0 mg total oxalates/100 g FW). Soluble oxalates were 74% of the total oxalate content of the young and old leaves.     

Ultraviolet light and Ultrasound as non-thermal treatments for the inactivation of microorganisms in fresh ready-to-eat foods

The effects of two non thermal disinfection processes, Ultraviolet light (UV 254 nm) and Ultrasound (US) on the inactivation of bacteria and color in two freshly cut produces (lettuce and strawberry) were investigated. The main scope of this work was to study the efficacy of UV and US on the decontamination of inoculated lettuce and strawberries with a cocktail of four bacteria, Escherichia coli, Listeria innocua, Salmonella Enteritidis and Staphylococcus aureus. Treatment of lettuce with UV reduced significantly the population of E. coli, L. innocua, S. Enteritidis and S. aureus by 1.75, 1.27, 1.39 and 1.21 log CFU/g, respectively. Furthermore, more than a 2-log CFU/g reduction of E. coli and S. Enteritidis was achieved with US. In strawberries, UV treatment reduced bacteria only by 1–1.4 log CFU/g. The maximum reductions of microorganisms, observed in strawberries after treatment with US, were 3.04, 2.41, 5.52 and 6.12 log CFU/g for E. coli, S. aureus, S. Enteritidis and L. innocua, respectively. Treatment with UV and US, for time periods (up to 45 min) did not significantly (p > 0.05) change the color of lettuce or strawberry. Treatment with UV and US reduced the numbers of selected inoculated bacteria on lettuce and strawberries, which could be good alternatives to other traditional and commonly used technologies such as chlorine and hydrogen peroxide solutions for fresh produce industry. These results suggest that UV and US might be promising, non-thermal and environmental friendly disinfection technologies for freshly cut produce.     

Role of O-antigen on the Escherichia coli O157:H7 cells hydrophobicity, charge and ability to attach to lettuce

Environmental factors encountered during growing and harvesting may contribute to Escherichia coli O157:H7 contamination of lettuce. Limited nutrients and extended exposure to water may cause E. coli O157:H7 to shed its O antigen. Absence of the O157-polysaccharide antigen could affect the cell's physicochemical properties (hydrophobicity and cell charge) and ultimately influence its attachment to surfaces. The objectives of this study were to evaluate the effect of the E. coli O157:H7 O-antigen on the cell's overall hydrophobicity, charge and ability to attach to cut edge and whole leaf iceberg lettuce surfaces. Three strains of E. coli O157:H7 (86-24 wild type; F-12, mutant lacking the O-antigen and pRFBE, plasmid for O157 gene reintroduced) were examined for their hydrophobicity, overall charge and ability to attach to lettuce. Overall, E. coli O157:H7 attached at higher levels to cut surfaces over whole leaf surfaces (P = 0.008) for all strains and treatments. Additionally, the strain lacking the O-antigen (F12) — attached significantly less to lettuce (P = 0.015) than the strains expressing the antigen (WT and pRFBE). Cells lacking the O antigen (strain F-12) were also significantly more hydrophobic than strains 86-24 or pRFBE (P ≤ 0.05). Surface charge differed among the strains tested (P ≤ 0.05); however, it did not appear to influence bacterial attachment to lettuce surfaces. The charge was not fully restored in the pRFBE strain (expression of O-antigen reintroduced), therefore, no conclusions can be made pertaining to the effect of charge on attachment in this study. Results indicate that E. coli O157:H7 cells which lack the O-antigen have greater hydrophobicity and attach at lower concentrations than cells expressing the O-antigen, to iceberg lettuce surfaces.     

Determination of water quality variables, endotoxin concentration, and Enterobacteriaceae concentration and identification in southern High Plains dairy lagoons

The objectives of this study were to determine the concentration of endotoxin, determine 20 water quality variables, and identify and enumerate fungal and bacterial pathogens from United States southern High Plains dairy lagoons and control lakes during summer and winter. Water samples were collected in triplicate from the north, south, east, and west quadrants of each body of water. The mean (± SEM) winter dairy lagoon endotoxin concentration was significantly higher (9,678 ± 1,834 ng/mL) than the summer concentration (3,220 ± 810 ng/mL). The mean endotoxin concentration of the 2 control lakes (summer: 58.1 ± 8.8 ng/mL; winter: 38.6 ± 4.2 ng/mL) was significantly less than that of the dairy lagoons. Two hundred-one Salmonella enterica spp. isolates were identified, 7 serovars were recovered from the dairy lagoons, and 259 Salmonella ssp. were identified from 5 other dairy locations (milk barn, ditch effluent, settling basin, feed alley pad flush, and center pivots). Twenty-eight Salmonella spp. were identified from center pivot water. Escherichia coli O157:H7 pathogens were isolated from other dairy locations but not from lagoons. Neither Salmonella spp. nor E. coli O157:H7 were identified from control lakes. Enterobacteriaceae opportunistic pathogens were isolated from both dairies and control lakes. Important mesophilic and thermophilic catabolic (to manure biosolids) fungal isolates were identified from dairy effluent locations, but no thermophilic fungal isolates were cultured from the control lakes. Adequate curing of green forage following center pivot irrigation is important to kill lagoon water enteric pathogens, even though the lagoon water is mixed with fresh water. Recirculating lagoon water to flush the feed alley pad, where cows stand while eating, to remove manure and using lagoon water to abate dairy dust in loafing pens and unimproved dairy roads is inconsistent with good environmental practice management.     

Salmonella on feces,hides and carcasses in beef slaughter facilities in Venezuela

This study determined Salmonella prevalence at different stages during the slaughtering in three beef slaughter plants (A, B and C) located in the western region of Venezuela (Zulia and Lara states). Each facility was visited three times at monthly intervals, from the months October through December of 2006. Samples were collected from hides (n = 80), fecal grabs (n = 80) and carcasses (n = 80) at the phases of pre-evisceration, after-evisceration and pre-cooler at three sampling sites on the animals (rump, flank and brisket). Salmonella prevalence was higher on hides (36.3%) than on feces (13.8%) (P < 0.05). Differences among slaughter plants for overall Salmonella prevalence were observed (P = 0.001; A: 3.5%, B: 11.1%, C: 4.4%). From the isolated strains, Salmonella enterica subspecies enterica ser. Saintpaul, Salmonella ser. Javiana and Salmonella ser. Weltevreden were identified. Cattle feces and hides might be considered as important sources of Salmonella for carcass contamination at different slaughter stages. The presence of potentially pathogenic Salmonella serotypes at the slaughtering stages is an evidence of the circulation of this pathogen in the food environment; its presence could increase consumers' risks of infection if proper food handling and preparation techniques are not followed. These data should serve as a baseline for future comparisons in Salmonella prevalence on beef carcasses to be used by the government and industry in order to establish preventive measures and to better address the risks of Salmonella contamination.     

Assessment of in vitro antioxidant capacity and polyphenolic composition of selected medicinal herbs from Leguminosae family in Peninsular Malaysia

Determination of total phenolic (TPC), flavonoid and anthocyanin contents, and various antioxidant activities (2,2-diphenyl-1-picrylhydrazyl radical scavenging, ferric reducing power, ferrous ion chelating and lipid peroxidation inhibition) of leaves and flowers of Bauhinia kockiana, Caesalpinia pulcherrima and Cassia surattensis were performed in this study. The B. kockiana flower was found to possess the highest TPC (8280 ± 498 mg GAE/100 g), free radical scavenging activity (ascorbic acid equivalents 14,600 ± 2360 mg AA/100 g) and reducing ability (72.4 ± 8.7 mg GAE/g). Rutin and chlorogenic acid were detected in the plants, where the C. pulcherrima leaf contained the highest amount of rutin (669 ± 26 mg/100 g), while minute amounts of chlorogenic acid were detected in C. surattensis leaf (9.13 ± 0.44 mg/100 g). The C. pulcherrima leaf displayed the highest ferrous ion chelating and lipid peroxidation inhibition activities. Positive correlation was observed between TPC and various antioxidant activities.     

Microbiological quality of fresh lettuce from organic and conventional production

Previously there was no available information on the levels of indicator bacteria and the prevalence of pathogens in fresh lettuce grown in organic and conventional farms in Spain. A total of 72 lettuce samples (18 farms for 4 repetitions each) for each type of the agriculture were examined in order to assess the bacteriological quality of the lettuces, in particular the prevalence of selected pathogens. The lettuce samples were analyzed for the presence of aerobic mesophilic, psychrotrophic microorganisms, yeasts and moulds, Enterobacteriaceae, mesophilic lactic acid bacteria, Pseudomonas spp. and presumptive Escherichia coli, Salmonella spp. and Listeria monocytogenes. The mean aerobic mesophilic counts (AM) were 6.35 ± 0.69 log₁₀ cfu g⁻¹ and 5.67 ± 0.80 log₁₀ cfu g⁻¹ from organic and conventional lettuce, respectively. The mean counts of psychrotrophic microorganisms were 5.82 ± 1.01 log₁₀ cfu g⁻¹ and 5.41 ± 0.92 log₁₀ cfu g⁻¹ from organic and conventional lettuce, respectively. Yeasts and moulds (YM) mean counts were 4.74 ± 0.83 log₁₀ cfu g⁻¹ and 4.21 ± 0.96 log₁₀ cfu g⁻¹ from organic and conventional lettuce, respectively. Lactic acid bacteria (LAB) were present in low numbers and the mean counts were 2.41 ± 1.10 log₁₀ cfu g⁻¹ and 1.99 ± 0.91 log₁₀ cfu g⁻¹ from organic and conventional lettuce, respectively. Pseudomonas spp. mean counts were 5.49 ± 1.37 log₁₀ cfu g⁻¹ and 4.98 ± 1.26 log₁₀ cfu g⁻¹ in organic and conventional lettuce, respectively. The mean counts for Enterobacteriaceae were 5.16 ± 1.01 log₁₀ cfu g⁻¹ and 3.80 ± 1.53 log₁₀ cfu g⁻¹ in organic and conventional lettuce, respectively. E. coli was detected in 22.2% (16 samples) of organic lettuce and in 12.5% (9 samples) of conventional lettuce. None of the lettuce samples was positive for E. coli O157:H7, L. monocytogenes and Salmonella spp. From the samples analyzed by principal component analysis (PCA) a pattern with two different groups (conventional and organic) can be observed, being the highest difference between both kinds of samples the Enterobacteriaceae count.     

Folate contents of some selected Fijian foods using tri-enzyme extraction method

Folic acid and total folate contents of 18 common foods in the Fijian diet were assayed. Foods were purchased from the central and eastern parts of Viti Levu in Fiji. Tri-enzyme treatment was performed to release bound folates using protease and α-amylase, with chicken pancreas as the conjugase. The highest total folate content was recorded for egg yolk (Gallus domesticus) at 256 μg/100 g, followed by long beans (Vigna sesquipedalis) which contained 130 μg/100 g of total folate (fresh weight basis). The local leafy vegetable called Bele (Abelmoschus manihot) and the Drumstick leaves (Moringa oleifera) available in Fiji also had high total folate contents, above 100 μg/100 g (fresh weight basis). For the 18 foods studied, the content of folic acid ranged from 3 to 189 μg/100 g and the total folate content was in the range of 3–256 μg/100 g, indicating a very wide range of folate content in the foods studied.     

Use of high-concentration-short-time chlorine dioxide gas treatments for the inactivation of Salmonella enterica spp. inoculated onto Roma tomatoes

Salmonella outbreaks have been recently linked to the consumption of fresh tomatoes. Thus, there is a need to develop systems that reduce the risk of microbial contamination to increase product shelf-life and keep fresh fruit attributes. The objectives of this study were to evaluate high-concentration-short-time chlorine dioxide gas treatments effects on Salmonella-inoculated Roma tomatoes and determine the optimal treatment conditions for microbial inactivation and shelf-life extension. Effects of ClO₂ concentration (2, 5, 8 and 10 mg/l) and exposure time (10, 30, 60, 120 and 180 s) on inoculated Roma tomatoes were studied. Salmonella enterica strains, serotype Montevideo, Javiana and Baildon, were used to experimentally inoculate the food product. After ClO₂ treatments, tomatoes were stored at room temperature for 28 days. Inherent microbial population, change in tomato color, and chlorine dioxide gas residuals were evaluated. ANOVA analysis showed that both ClO₂ concentration and treatment time were significant (p < 0.01) for Salmonella inactivation. Surviving Salmonella populations of 3.09, 2.17 and 1.16 log CFU/cm² were obtained treating tomatoes with 8 mg/l ClO₂ for 60 s, 10 mg/l ClO₂ for 120 s, and 10 mg/l for 180 s, respectively (initial Salmonella population: 6.03 ± 0.11 log CFU/cm²). The selected treatments significantly reduced background microflora (p < 0.05), while fruit color and residual contents were not significantly different (p > 0.05), as compared to the control. Results suggest the potential for high-concentration-short-time treatments ClO₂ gas as an effective pathogen inactivation technology for large-scale produce packing operations.     

Inactivation mechanism of Salmonella Typhimurium on the surface of lettuce and physicochemical quality assessment of samples treated by micro-plasma discharged water

The potential of micro-plasma discharged water (m-PDW) treatment on reduction of Salmonella Typhimurium ATCC 14028 from the surface of lettuce leaves was evaluated. The surface of iceberg lettuce and red leaf lettuce pieces (3 cm × 3 cm) was inoculated with a S. Typhimurium suspension. The measured S. Typhimurium reductions were 3.0-log CFU/piece and 2.6-log CFU/piece for iceberg lettuce and red leaf lettuce, respectively for a 3-min treatment. The overall reductions measured for the levels of kaempferol and quercetin in both types of lettuce were within the experimental error for a 3-min plasma treatment. There was no significant change in color (p > 0.05) for plasma operations of the same duration. Leaching of water contributed to a greater extent towards physiochemical changes such as color constraints, kaempferol and quercetin content occurred for the treated lettuce samples, although the overall physicochemical quality changes measured in this study remains in the satisfactory level.     

Characterization of polyphenol oxidase from butter lettuce (Lactuca sativa var. capitata L.)

Polyphenol oxidase (PPO) was isolated from butter lettuce (Lactuca sativa var. capitata L.) grown in Poland and its biochemical characteristic were studied. PPO from butter lettuce showed a higher affinity to 4-methylcatechol than to catechol. The K_M and V_max values were: 3.20 ± 0.01 mM and 4081 ± 8 U/ml min⁻¹ for catechol and 1.00 ± 0.09 mM and 5405 ± 3 U/ml min⁻¹ for 4-methylcatechol. The optimum pHs of the enzyme were found to be 5.5 using catechol and 6.8 using 4-methylcatechol as substrate. The enzyme had a temperature optimum of 35 °C. The enzyme was relatively stable at 30 °C and 40 °C. The times required for 50% inactivation of activity at 50 °C, 60 °C and 70 °C were found to be about 30, 20 and 5 min, respectively. Inhibitors used for investigation in this study were placed in relative order of inhibition: p-hydroxybenzoic acid > glutathione ≈ ascorbic acid > l-cysteine > EDTA > citric acid. The enzyme eluted in the chromatographic separations was analyzed electrophoretically under denaturating conditions. The analysis revealed a single band on the SDS–PAGE which corresponded to a molecular weight of 60 kDa.