Efficiency of zinc in alleviating cadmium toxicity in hydroponically grown lettuce (Lactuca sativa L. cv. Ferdos).

BACKGROUND: A study on photosynthetic and enzyme activity changes and mineral content in lettuce under cadmium stress has been conducted in a greenhouse, utilizing the modulated effect of zinc (Zn) application in the nutrient solution on lettuce. Zn is a micronutrient that plays an essential role in various critical plant processes. Accordingly, three concentrations of Zn (0.022, 5, and 10 mg L- 1) were applied to hydroponically grown lettuce (Lactuca sativa L. cv. Ferdos) under three concentrations of Cd toxicity (0, 2.5, and 5 mg L- 1). RESULTS: The results showed that along with increasing concentrations of zinc in the nutrient solution, growth traits such as plant performance, chlorophyll index (SPAD), minimum fluorescence (F0), leaf zinc content (Zn), leaf and root iron (Fe) content, manganese (Mn), copper (Cu), and cadmium increased as well. The maximum amounts of chlorophyll a (33.9 mg g- 1FW), chlorophyll b (17.3 mg g- 1FW), carotenoids (10.7 mg g- 1FW), maximum fluorescence (Fm) (7.1), and variable fluorescence (Fv) (3.47) were observed in the treatment with Zn without Cd. Along with an increase in Cd concentration in the nutrient solution, the maximum amounts of leaf proline (5.93 mmol g- 1FW), malondialdehyde (MDA) (0.96 µm g- 1FW), hydrogen peroxide (H2O2) (22.1 µm g- 1FW), and superoxide dismutase (SOD) (90.3 Unit mg- 1 protein) were recorded in lettuce treated with 5 mg L- 1 of Cd without Zn. Additionally, the maximum activity of leaf guaiacol peroxidase (6.46 Unit mg- 1 protein) was obtained with the application of Cd at a 5 mg L- 1 concentration. CONCLUSIONS: In general, an increase in Zn concentration in the nutrient solution decreased the absorption and toxicity of Cd in lettuce leaves, as demonstrated in most of the measured traits. These findings suggest that supplementing hydroponic nutrient solutions with zinc can mitigate the detrimental effects of cadmium toxicity on lettuce growth and physiological processes, offering a promising strategy to enhance crop productivity and food safety in cadmium-contaminated environments.

Quinoa greens as a novel plant food: a review of its nutritional composition, functional activities, and food applications.

Quinoa (Chenopodium quinoa Willd) is widely regarded as a versatile pseudo-cereal native to the Andes Mountains in South America. It has gained global recognition as a superfood due to its rich nutritional profile. While quinoa grains are well-known, there is an undiscovered potential in quinoa greens, such as sprouts, leaves, and microgreens. These verdant parts of quinoa are rich in a diverse array of essential nutrients and bioactive compounds, including proteins, amino acids, bioactive proteins, peptides, polyphenols, and flavonoids. They have powerful antioxidant properties, combat cancer, and help prevent diabetes. Quinoa greens offer comparable or even superior benefits when compared to other sprouts and leafy greens, yet they have not gained widespread recognition. Limited research exists on the nutritional composition and biological activities of quinoa greens, underscoring the necessity for thorough systematic reviews in this field. This review paper aims to highlight the nutritional value, bioactivity, and health potential of quinoa greens, as well as explore their possibilities within the food sector. The goal is to generate interest within the research community and promote further exploration and wider utilization of quinoa greens in diets. This focus may lead to new opportunities for enhancing health and well-being through innovative dietary approaches.

It takes two to attach - endo-1,3-ß-d-glucanase as a potential receptor of mannose-independent, FimH-dependent Salmonella Typhimurium binding to spinach leaves.

Currently, fresh, unprocessed food has become a relevant element of the chain of transmission of enteropathogenic infections. To survive on a plant surface and further spread the infections, pathogens like Salmonella have to attach stably to the leaf surface. Adhesion, driven by various virulence factors, including the most abundant fim operon encoding type 1 fimbriae, is usually an initial step of infection, preventing physical removal of the pathogen. Adhesion properties of Salmonella's type 1 fimbriae and its FimH adhesin were investigated intensively in the past. However, there is a lack of knowledge regarding its role in interaction with plant cells. Understanding the mechanisms and structures involved in such interaction may facilitate efforts to decrease the risk of contamination and increase fresh food safety. Here, we applied Salmonella genome site-directed mutagenesis, adhesion assays, protein-protein interactions, and biophysics methods based on surface plasmon resonance to unravel the role of FimH adhesin in interaction with spinach leaves. We show that FimH is at least partially responsible for Salmonella binding to spinach leaves, and this interaction occurs in a mannose-independent manner. Importantly, we identified a potential FimH receptor as endo-1,3-ß-d-Glucanase and found that this interaction is strong and specific, with a dissociation constant in the nanomolar range. This research advances our comprehension of Salmonella's interactions with plant surfaces, offering insights that can aid in minimizing contamination risks and improving the safety of fresh, unprocessed foods.

Fates of attached E. coli o157:h7 on intact leaf surfaces revealed leafy green susceptibility.

Leafy greens, especially lettuce, are repeatedly linked to foodborne outbreaks. This paper studied the susceptibility of different leafy greens to human pathogens. Five commonly consumed leafy greens, including romaine lettuce, green-leaf lettuce, baby spinach, kale, and collard, were selected by their outbreak frequencies. The behavior of E. coli O157:H7 87-23 on intact leaf surfaces and in their lysates was investigated. Bacterial attachment was positively correlated with leaf surface roughness and affected by the epicuticular wax composition. At room temperature, E. coli O157:H7 had the best growth potentials on romaine and green-leaf lettuce surfaces. The bacterial growth was positively correlated with stomata size and affected by epicuticular wax compositions. At 37 °C, E. coli O157:H7 87-23 was largely inhibited by spinach and collard lysates, and it became undetectable in kale lysate after 24 h of incubation. Kale and collard lysates also delayed or partially inhibited the bacterial growth in TSB and lettuce lysate at 37 °C, and they sharply reduced the E. coli O157:H7 population on green leaf lettuce at 4 °C. In summary, the susceptibility of leafy greens to E. coli O157:H7 is determined by a produce-specific combination of physiochemical properties and temperature.

Novel RP-HPLC method for estimation of a newly developed combination of tizanidine and etoricoxib in rat plasma: Eight criteria for greens evaluation.

A novel environmentally friendly reversed-phase high-performance liquid chromatography (RP-HPLC) method has been effectively validated for simultaneously measuring a prospective conjunction of tizanidine (TIZ) and etoricoxib (ETC), the combined medicine, in rat plasma. The technique employs diclofenac potassium as the internal standard, guaranteeing dependable and precise outcomes. This study aimed to assess the impact of the suggested combination therapy on treating inflammation resulting from rheumatoid arthritis (RA) in a rat model. The procedure was performed using an Agilent series 1200 model HPLC apparatus. The chromatographic conditions consist of isocratic elution mode, C18 column with dimensions of 150 mm × 4.6 mm × 5 µm, flow rate of 1.5 mL/min, wavelength of 230 nm, temperature of 50°C, and injection volume of 10 µL. The elution was performed using a mobile phase consisting of a phosphate buffer with a pH of 3.5 and acetonitrile in a ratio of 80:20 v/v. Calibration curves were conducted for TIZ and ETC within the 1-50 µg/mL range, demonstrating linear trends with R2 values over 0.999. The effectiveness and eco-friendliness of the proposed method were evaluated using eight separate environmentally conscious metrics. The addition of TIZ and ETC to arthritic rodents amplified these effects significantly. Furthermore, TIZ and ETC significantly reduced serum levels in arthritic rodents, and safety investigations revealed normal complete blood count, liver, and renal functions. TIZ and ETC appear to have antiarthritic, anti-inflammatory, and safe combinations, making them viable future treatment options for RA that are also safe and efficacious. Following validation by United States Food and Drug Administration (US-FDA) rules, all goods met the criteria.

Predictive Population Dynamics of Escherichia coli O157:H7 and Salmonella enterica on Plants: a Mechanistic Mathematical Model Based on Weather Parameters and Bacterial State.

Weather affects key aspects of bacterial behavior on plants but has not been extensively investigated as a tool to assess risk of crop contamination with human foodborne pathogens. A novel mechanistic model informed by weather factors and bacterial state was developed to predict population dynamics on leafy vegetables and tested against published data tracking Escherichia coli O157:H7 (EcO157) and Salmonella enterica populations on lettuce and cilantro plants. The model utilizes temperature, radiation, and dew point depression to characterize pathogen growth and decay rates. Additionally, the model incorporates the population level effect of bacterial physiological state dynamics in the phyllosphere in terms of the duration and frequency of specific weather parameters. The model accurately predicted EcO157 and S. enterica population sizes on lettuce and cilantro leaves in the laboratory under various conditions of temperature, relative humidity, light intensity, and cycles of leaf wetness and dryness. Importantly, the model successfully predicted EcO157 population dynamics on 4-week-old romaine lettuce plants under variable weather conditions in nearly all field trials. Prediction of initial EcO157 population decay rates after inoculation of 6-week-old romaine plants in the same field study was better than that of long-term survival. This suggests that future augmentation of the model should consider plant age and species morphology by including additional physical parameters. Our results highlight the potential of a comprehensive weather-based model in predicting contamination risk in the field. Such a modeling approach would additionally be valuable for timing field sampling in quality control to ensure the microbial safety of produce. IMPORTANCE Fruits and vegetables are important sources of foodborne disease. Novel approaches to improve the microbial safety of produce are greatly lacking. Given that bacterial behavior on plant surfaces is highly dependent on weather factors, risk assessment informed by meteorological data may be an effective tool to integrate into strategies to prevent crop contamination. A mathematical model was developed to predict the population trends of pathogenic E. coli and S. enterica, two major causal agents of foodborne disease associated with produce, on leaves. Our model is based on weather parameters and rates of switching between the active (growing) and inactive (nongrowing) bacterial state resulting from prevailing environmental conditions on leaf surfaces. We demonstrate that the model has the ability to accurately predict dynamics of enteric pathogens on leaves and, notably, sizes of populations of pathogenic E. coli over time after inoculation onto the leaves of young lettuce plants in the field.

Melissa officinalis essential oil as an antimicrobial agent against Listeria monocytogenes in watermelon juice.

The burden of foodborne illness has a negative effect on public health, but also in countries' economy. Melissa officinalis is an aromatic plant known for its biological properties, including antioxidant and antimicrobial effects. This work highlighted M. officinalis essential oil's antioxidant potential and antimicrobial activity against L. monocytogenes, presenting a bactericidal action and being able to inhibit some virulence attributes, such as biofilm formation. The pre-exposure of the bacterium to subinhibitory levels of essential oil (0.125 µL/mL) did not induce high tolerance to stresses (such as high temperature, low pH, osmotic stress and desiccation) or cross-resistance with antibiotics, while not modifying the invasion ability to Caco-2 cells. When applied in food model media (lettuce, chicken and milk) and watermelon juice, the essential oil showed to have antimicrobial activity in a lettuce leaf model medium, further diminishing L. monocytogenes contamination and inhibiting the natural microbiota present in watermelon juice. M. officinalis essential oil shows potential to be used as control of L. monocytogenes in watermelon juice, while increasing the food's microbial shelf life.

Weather factors, soil microbiome, and bacteria-fungi interactions as drivers of the epiphytic phyllosphere communities of romaine lettuce.

Lettuce is associated with seasonal outbreaks of Shiga toxin-producing Escherichia coli (STEC) infections. Little is known about how various biotic and abiotic factors affect the lettuce microbiome, which in turn impacts STEC colonization. We characterized the lettuce phyllosphere and surface soil bacterial, fungal, and oomycete communities at harvest in late-spring and -fall in California using metagenomics. Harvest season and field type, but not cultivar, significantly influenced the microbiome composition of leaves and surface soil near plants. Phyllosphere and soil microbiome compositions were correlated with specific weather factors. The relative abundance of Enterobacteriaceae, but not E. coli, was enriched on leaves (5.2%) compared to soil (0.4%) and correlated positively with minimum air temperature and wind speed. Co-occurrence networks revealed seasonal trends in fungi-bacteria interactions on leaves. These associations represented 39%-44% of the correlations between species. All significant E. coli co-occurrences with fungi were positive, while all negative associations were with bacteria. A large proportion of the leaf bacterial species was shared with those in soil, indicating microbiome transmission from the soil surface to the canopy. Our findings provide new insight into factors that shape lettuce microbial communities and the microbial context of foodborne pathogen immigration events in the lettuce phyllosphere.

Growth models for Salmonella, E. coli O157:H7 and L. monocytogenes give different predictions for pathogen growth in cut leafy greens transportation, but are consistent in identifying higher risk conditions.

Leafy greens are frequently implicated in foodborne disease outbreaks and cut-leafy greens are a food that requires time and temperature control for safety. Predictive microbiology uses mathematical models to predict the growth of bacteria based on environmental conditions. The objective of our study was to compare published square root growth models for Salmonella (n = 6), pathogenic E. coli (n = 6) and Listeria monocytogenes (n = 4) using real world transport temperature data. Data from trucks transporting fresh-cut leafy greens during cross-country shipments were used as temperature inputs to the models. Bacterial growth was computed using the temperatures from each probe in every truck over the duration of transit, which resulted in 12-18 growth predictions per truck for each model. Each model generally gave significantly different predictions than other models for the same organism. The exception was for the two Salmonella models predicting the least growth and the two Salmonella models predicting the most growth which gave predictions that were not significantly different. Although different models tended to give different predictions, their ability to rank risk by truck was generally consistent across models. While absolute risk might be dependent upon choice of model, relative risk is independent of model choice.

Quantitative risk assessment-epidemic curve prediction model for leafy green outbreak investigation.

The objective of this study was to leverage quantitative risk assessment to investigate possible root cause(s) of foodborne illness outbreaks related to Shiga toxin-producing Escherichia coli O157:H7 (STEC O157) infections in leafy greens in the United States. To this end, we developed the FDA leafy green quantitative risk assessment epidemic curve prediction model (FDA-LG QRA-EC) that simulated the lettuce supply chain. The model was used to predict the number of reported illnesses and the epidemic curve associated with lettuce contaminated with STEC O157 for a wide range of scenarios representing various contamination conditions and facility processing/sanitation practices. Model predictions were generated for fresh-cut and whole lettuce, quantifying the differing impacts of facility processing and home preparation on predicted illnesses. Our model revealed that the timespan (i.e., number of days with at least one reported illness) and the peak (i.e., day with the most predicted number of reported illnesses) of the epidemic curve of a STEC O157-lettuce outbreak were not strongly influenced by facility processing/sanitation practices and were indications of contamination pattern among incoming lettuce batches received by the facility or distribution center. Through comparisons with observed number of illnesses from recent STEC O157-lettuce outbreaks, the model identified contamination conditions on incoming lettuce heads that could result in an outbreak of similar size, which can be used to narrow down potential root cause hypotheses.

Phage Therapy Against Pathogenic Escherichia coli (O104:H4, O157:H7, and O26) Strains in Irrigation Water During Garden Cress (Lepidium sativum Linn.) Vegetation.

Fresh produce outbreaks have increased worldwide. Foodborne pathogens are transmitted mostly by contaminated water, and elimination is harder after the transmission. To eliminate pathogens in fresh produce, chemical prevention methods, including chlorine, can be used. However, the usage of chemicals poses a risk to human health, as well as environmental health. Therefore, alternative prevention methods that can be applied in the field should be investigated. This study aims to investigate an alternative method to prevent the pathogenic Escherichia coli strain O26 and Shiga toxin-producing strains O104:H4 and O157:H7 on freshly consumed garden cresses. In this study, garden cresses were treated with bacteriophages after becoming contaminated with pathogenic E. coli strains during growth. After 30 days, the leaves were collected and tested for the presence of E. coli. Its adherence on the leaf surface was investigated with scanning electron microscope (SEM). Although there were significant reductions in both total and biofilm-forming E. coli counts in pathogenic E. coli strain O26 and Shiga toxin-producing strains O104:H4 and O157:H7, which is also confirmed with the SEM images, the counts were not lowered to levels permitted by the EU. Therefore, results showed that phage therapy against pathogenic E. coli strains may be used as a biocontrol agent in combination with additional control measure.

A Validated Preharvest Sampling Simulation Shows that Sampling Plans with a Larger Number of Randomly Located Samples Perform Better than Typical Sampling Plans in Detecting Representative Point-Source and Widespread Hazards in Leafy Green Fields.

Commercial leafy greens customers often require a negative preharvest pathogen test, typically by compositing 60 produce sample grabs of 150 to 375 g total mass from lots of various acreages. This study developed a preharvest sampling Monte Carlo simulation, validated it against literature and experimental trials, and used it to suggest improvements to sampling plans. The simulation was validated by outputting six simulated ranges of positive samples that contained the experimental number of positive samples (range, 2 to 139 positives) recovered from six field trials with point source, systematic, and sporadic contamination. We then evaluated the relative performance between simple random, stratified random, or systematic sampling in a 1-acre field to detect point sources of contamination present at 0.3% to 1.7% prevalence. Randomized sampling was optimal because of lower variability in probability of acceptance. Optimized sampling was applied to detect an industry-relevant point source [3 log(CFU/g) over 0.3% of the field] and widespread contamination [-1 to -4 log(CFU/g) over the whole field] by taking 60 to 1,200 sample grabs of 3 g. More samples increased the power of detecting point source contamination, as the median probability of acceptance decreased from 85% with 60 samples to 5% with 1,200 samples. Sampling plans with larger total composite sample mass increased power to detect low-level, widespread contamination, as the median probability of acceptance with -3 log(CFU/g) contamination decreased from 85% with a 150-g total mass to 30% with a 1,200-g total mass. Therefore, preharvest sampling power increases by taking more, smaller samples with randomization, up to the constraints of total grabs and mass feasible or required for a food safety objective. IMPORTANCE This study addresses a need for improved preharvest sampling plans for pathogen detection in leafy green fields by developing and validating a preharvest sampling simulation model, avoiding the expensive task of physical sampling in many fields. Validated preharvest sampling simulations were used to develop guidance for preharvest sampling protocols. Sampling simulations predicted that sampling plans with randomization are less variable in their power to detect low-prevalence point source contamination in a 1-acre field. Collecting larger total sample masses improved the power of sampling plans in detecting widespread contamination in 1-acre fields. Hence, the power of typical sampling plans that collect 150 to 375 g per composite sample can be improved by taking more, randomized smaller samples for larger total sample mass. The improved sampling plans are subject to feasibility constraints or to meet a particular food safety objective.

Rapid detection of Salmonella enterica in leafy greens by a novel DNA microarray-based PathogenDx system.

The diverse matrices pose great challenges for rapid detection of low Salmonella level (<10 CFU) in fresh produce. The applicability of microarray-based PathogenDx system for detecting low contamination of Salmonella Newport from leafy greens was evaluated. A pre-PCR preparation protocol including enrichment in universal pre-enrichment broth for 3 h followed by sample concentration using an InnovaPrep bio-concentrator or 6 h enrichment without a concentration step was used for detecting S. Newport from leafy greens with initial inoculum level at ∼6 CFU/25 g. Among 205 samples tested, 98%, 93%, 76%, and 60% of Romaine lettuce, Iceberg lettuce, kale, and spinach samples were tested positive after 3 h of enrichment with sample concentration. After 6 h of enrichment, 100%, 98%, 90%, and 82% of Romaine lettuce, Iceberg lettuce, kale, and spinach samples were positive. The samples were parallelly tested by the FDA bacterial analytical manual (BAM) method and 100% of spiked produce samples were tested positive. The overall analysis time of this methodology was between 8 and 11 h, including all pre-enrichment and concentration steps, in contrast to 4-5 days required for BAM method. The system correctly differentiated all 108 Salmonella strains and 35 non-Salmonella strains used in the study. This novel microarray approach provides a rapid method for detecting Salmonella in leafy greens.

Evaluation of Potential Impacts of Free Chlorine during Washing of Fresh-Cut Leafy Greens on Escherichia coli O157:H7 Cross-Contamination and Risk of Illness.

Addition of chlorine-based antimicrobial substances to fresh-cut leafy green wash water is done to minimize microbial cross-contamination during processing. We developed the FDA Leafy Green Risk Assessment Model (FDA-LGRAM) to quantify the impact of free chlorine concentration in wash water during fresh-cut lettuce processing on the extent of water-mediated cross-contamination between shredded lettuce and the associated risk of illness due to exposure to Escherichia coli O157:H7. At different contamination prevalence and levels of E. coli O157:H7 on incoming lettuce heads, the model compared the predicted prevalence of contaminated fresh-cut lettuce packages and the risk of illness per serving between: (1) a scenario where fresh-cut lettuce was packaged without washing; and (2) scenarios involving washing fresh-cut lettuce with different levels of free chlorine (0 ppm, 5 ppm, 10 ppm, 15 ppm, and 20 ppm) prior to packaging. Our results indicate that the free chlorine level in wash water has a substantial impact on the predicted prevalence of contaminated fresh-cut lettuce packages and the risk of illness associated with E. coli O157:H7 in fresh-cut lettuce. Results showed that the required level of free chlorine that can minimize water-mediated cross-contamination and reduce the corresponding risk of illness depended on contamination prevalence and levels of E. coli O157:H7 on incoming lettuce heads. Our model also indicated that the pathogen inactivation rate in wash water via free chlorine was a key model parameter that had a significant impact on the extent of cross-contamination during washing and the predicted associated risk of illness.

Effect of Acidic Electrolysed Water and Pulsed Light Technology on the Sensory, Morphology and Bioactive Compounds of Pennywort (Centella asiatica L.) Leaves.

Pennywort (Centella asiatica) is a herbaceous vegetable that is usually served in the form of fresh-cut vegetables and consumed raw. Fresh-cut vegetables are in high demand as they offer convenience, have fresh-like quality and are potentially great for therapeutic applications. However, it could be the cause of foodborne outbreaks. Pulsed light is known as a decontamination method for minimally processed products. The aim of this study was to determine the influence of pulsed light in combination with acidic electrolysed water on the sensory, morphological changes and bioactive components in the leaves of pennywort during storage. A combination of soaking with acidic electrolysed water (AEW) at pH 2.5 and pulsed light (PL) treatment (1.5 J/cm2) was tested on the leaves of pennywort. After treatment, these leaves were refrigerated (4 ± 1 °C) for two weeks and evaluated on the basis of sensory acceptance, the visual appearance of the epidermal cell and bioactive compounds. In terms of sensorial properties, samples treated with the combined treatment were preferred over untreated samples. The combination of AEW and PL 1.5 J/cm2 was the most preferred in terms of purchasing and consumption criteria. Observations of the epidermal cells illustrated that PL treatment kept the cell structure intact. The bioactive phytocompounds found in the leaves of pennywort are mainly from the triterpene glycosides (asiaticoside, madecassoside, asiatic acid and madecassic acid) and are efficiently preserved by the combined treatment applied. In conclusion, the combination of acidic electrolysed water and pulsed light treatment is beneficial in retaining the sensory quality and bioactive compounds in the leaves of Pennywort during storage at 4 ± 1 °C.

In vitro gastrointestinal digestion studies on total phenols, flavonoids, anti-oxidant activity and vitamin C in freeze-dried vegetable powders.

In the present research study, the impact of digestion process on the levels of total phenols, flavonoids, vitamin C as well as anti-oxidant activity in freeze dried powders of mustard greens (MG) and roselle leaves (RL) was investigated. In addition, physicochemical and functional properties of MG and RL samples also evaluated. The digestion of freeze-dried vegetable powders was achieved through in vitro digestive procedure using various enzymes. From the study, it was observed that the digestion process increased the availability of phenols in both powders, where the digested vegetable powders possessed higher levels of total phenols and flavonoids. In contrast, the levels of vitamin C and anti-oxidant activity of vegetable powders (MG & RL) was found to be decreased minimally. Our research study suggests that in vitro digestion could enhance the TPC and TFC in mustard greens and roselle leaves. Therefore, MG and RL can be considered as a functional ingredient in the development of new products with better nutritional and functional characteristics. Further, the data on the physicochemical, functional and bioactive compounds in MG and RL may be used as reference for the enhancement of quality of products developed from MG and RL.

Leafy greens as a potential source of multidrug-resistant diarrhoeagenic Escherichia coli and Salmonella.

A continued rise in leafy green-linked outbreaks of disease caused by pathogenic Escherichia coli or Salmonella, particularly strains exhibiting multidrug resistance (MDR), has emerged as a major threat to human health and food safety worldwide. Thus, the present study was conducted to examine antimicrobial resistance, including MDR, in diarrhoeagenic E. coli (DEC) and Salmonella isolates obtained from leafy greens from rural and urban areas of India. Of the collected samples (830), 14.1 and 6.5% yielded 117 E. coli (40 DEC and 77 non-DEC) and 54 Salmonella isolates, respectively. Among the DEC pathotypes, enteroaggregative E. coli was the most prevalent (10.2 %), followed by enteropathogenic E. coli (9.4 %), enteroinvasive E. coli (7.6 %) and enterohemorrhagic E. coli (6.8 %). Antimicrobial susceptibility testing of all bacterial isolates with respect to drugs categorized as critically or highly important in both human and veterinary medicine revealed moderate to high (30-90%) resistance for amoxicillin/clavulanic acid, ampicillin, gentamycin and colistin, but relatively low resistance (>30 %) for ciprofloxacin, trimethoprim/sulfamethoxazole and fosfomycin. Notably, all DEC and more than 90% non-DEC or Salmonella isolates were found to be multidrug-resistant to drugs of both human and animal importance. Overall, the results of the present study suggest that leafy greens are potential reservoirs or sources of multidrug-resistant DEC and Salmonella strains in the rural or urban areas of India.

Effect of Plant Systemic Resistance Elicited by Biological and Chemical Inducers on the Colonization of the Lettuce and Basil Leaf Apoplast by Salmonella enterica.

Mitigation strategies to prevent microbial contamination of crops are lacking. We tested the hypothesis that induction of plant systemic resistance by biological (induced systemic resistance [ISR]) and chemical (systemic acquired resistance [SAR]) elicitors reduces endophytic colonization of leaves by Salmonella enterica serovars Senftenberg and Typhimurium. S. Senftenberg had greater endophytic fitness than S. Typhimurium in basil and lettuce. The apoplastic population sizes of serovars Senftenberg and Typhimurium in basil and lettuce, respectively, were significantly reduced approximately 10- to 100-fold by root treatment with microbial inducers of systemic resistance compared to H2O treatment. Rhodotorula glutinis effected the lowest population increases of S. Typhimurium in lettuce and S. Senftenberg in basil leaves, respectively 120- and 60-fold lower than those seen with the H2O treatment over 10 days postinoculation. Trichoderma harzianum and Pichia guilliermondii did not have any significant effect on S. Senftenberg in the basil apoplast. The chemical elicitors acidobenzolar-S-methyl and dl-ß-amino-butyric acid inhibited S. Typhimurium multiplication in the lettuce apoplast 10- and 2-fold, respectively, compared to H2O-treated plants. All ISR and SAR inducers applied to lettuce roots in this study increased leaf expression of the defense gene PR1, as did Salmonella apoplastic colonization in H2O-treated lettuce plants. Remarkably, both acidobenzolar-S-methyl upregulation and R. glutinis upregulation of PR1 were repressed by the presence of Salmonella in the leaves. However, enhanced PR1 expression was sustained longer and at greater levels upon elicitor treatment than by Salmonella induction alone. These results serve as a proof of concept that priming of plant immunity may provide an intrinsic hurdle against the endophytic establishment of enteric pathogens in leafy vegetables. IMPORTANCE Fruit and vegetables consumed raw have become an important vehicle of foodborne illness despite a continuous effort to improve their microbial safety. Salmonella enterica has caused numerous recalls and outbreaks of infection associated with contaminated leafy vegetables. Evidence is increasing that enteric pathogens can reach the leaf apoplast, where they confront plant innate immunity. Plants may be triggered for induction of their defense signaling pathways by exposure to chemical or microbial elicitors. This priming for recognition of microbes by plant defense pathways has been used to inhibit plant pathogens and limit disease. Given that current mitigation strategies are insufficient in preventing microbial contamination of produce and associated outbreaks, we investigated the effect of plant-induced resistance on S. enterica colonization of the lettuce and basil leaf apoplast in order to gain a proof of concept for the use of such an intrinsic approach to inhibit human pathogens in leafy vegetables.

Assessment of microbial risk during Australian industrial practices for Escherichia coli O157:H7 in fresh cut-cos lettuce: A stochastic quantitative approach.

Escherichia coli O157:H7 risk associated with the consumption of fresh cut-cos lettuce during Australian industrial practices was assessed. A probabilistic risk assessment model was developed and implemented in the @Risk software by using the Monte Carlo simulation technique with 1,000,000 iterations. Australian preharvest practices yielded predicted annual mean E. coli O157:H7 levels from 0.2 to -3.4 log CFU/g and prevalence values ranged from 2 to 6.4%. While exclusion of solar radiation from the baseline model yielded a significant increase in concentration of E. coli O157:H7 (-5.2 -log fold), drip irrigation usage, exclusion of manure amended soil and rainfall reduced E. coli O157:H7 levels by 7.4, 6.5, and 4.3-log fold, respectively. The microbial quality of irrigation water and irrigation type both had a significant effect on E. coli O157:H7 concentrations at harvest (p < 0.05). The probability of illness due to consumption of E. coli O157:H7 contaminated fresh cut-cos lettuce when water washing interventions were introduced into the processing module, was reduced by 1.4-2.7-log fold (p < 0.05). This study provides a robust basis for assessment of risk associated with E. coli O157:H7 contamination on fresh cut-cos lettuce for industrial practices and will assist the leafy green industry and food safety authorities in Australia to identify potential risk management strategies.

Recycling pyrolyzed organic waste from plant nurseries, rice production and shrimp industry as peat substitute in potting substrates.

Organic waste from greens of tomato plants, gardening substrate, rice husks and shrimp-derived chitin were pyrolyzed at 400 °C and 500 °C for 3 h, with the aim to elucidate the feasibility of using such products as replacement of peat in soilless gardening substrates. Characterization of the carbonized organic matter (COM) and the gardening substrate indicated that neither the peat nor the COMs provided the recommended levels of nutrients for the cultivation of tomato plants, although improvements could be obtained using COM/substrate mixtures. The toxicity thresholds for Zn were exceeded significantly by the COMs of the tomato greens and high boron levels were found for all the COMs except for those derived from chitin. In a 40-days pot experiment, germination and development of tomato seeds and plants (Solanum lycopersicum L.) were tested on COM/peat mixtures at 30%, 60% and 100% COM substitution rate. The lack of seed germination on the mixtures with COM from tomato greens is best explained with the high salinity of the COM. Best plant growth was obtained with COM from chitin at 60%, most likely because its high N content satisfied best the N-needs of the growing tomato plants without increasing the pH of the growing media. Moreover, an increase of water retention was evidenced for COM/substrate mixtures. Although the use of COM from chitin and rice husks showed promising results, the proposed recycling of organic waste from agriculture or fishery as soilless gardening substrate requires the development of formulations of COM/peat/and added nutrients with ready-to-use characteristics to increase its feasibility.