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Fusarium oxysporum (Schl.) f.sp. melonis, which causes muskmelon wilt disease, is a destructive filamentous fungal pathogen, attracting more attention to the search for effective fungicides against this pathogen. In particular, Silver nanoparticles (AgNPs) have strong antimicrobial properties and they are not easy to develop drug resistance, which provides new ideas for the prevention and control of muskmelon Fusarium wilt (MFW). This paper studied the effects of AgNPs on the growth and development of muskmelon, the control efficacy on Fusarium wilt of muskmelon and the antifungal mechanism of AgNPs to F. oxysporum. The results showed that AgNPs could inhibit the growth of F. oxysporum on the PDA and in the PDB medium at 100-200 mg/L and the low concentration of 25 mg/L AgNPs could promote the seed germination and growth of muskmelon seedlings and reduce the incidence of muskmelon Fusarium wilt. Further studies on the antifungal mechanism showed that AgNPs could impair the development, damage cell structure, and interrupt cellular metabolism pathways of this fungus. TEM observation revealed that AgNPs treatment led to damage to the cell wall and membrane and accumulation of vacuoles and vessels, causing the leakage of intracellular contents. AgNPs treatment significantly hampered the growth of mycelia in the PDB medium, even causing a decrease in biomass. Biochemical properties showed that AgNPs treatment stimulated the generation of reactive oxygen species (ROS) in 6 h, subsequently producing malondialdehyde (MDA) and increasing protective enzyme activity. After 6 h, the protective enzyme activity decreased. These results indicated that AgNPs destroy the cell structure and affect the metabolisms, eventually leading to the death of fungus., (© 2024. The Author(s).)

A novel mitovirus was identified in Fusarium oxysporum f. sp. melonis strain T-SD3 and designated as "Fusarium oxysporum mitovirus 3" (FoMV3). The virus was isolated from diseased muskmelon plants with the typical symptom of fusarium wilt. The complete genome of FoMV3 is 2269 nt in length with a predicted AU content of 61.40% and contains a single open reading frame (ORF) using the fungal mitochondrial genetic code. The ORF was predicted to encode a polypeptide of 679 amino acids (aa) containing a conserved RNA-dependent RNA polymerase (RdRp) domain with a molecular mass of 77.39 kDa, which contains six conserved motifs with the highly conserved GDD tripeptide in motif IV. The 5'-untranslated region (UTR) and 3'-UTR of FoMV3 were predicted to fold into stem-loop structures. BLASTp analysis revealed that the RdRp of FoMV3 shared the highest aa sequence identity (83.85%) with that of Fusarium asiaticum mitovirus 5 (FaMV5, a member of the family Mitoviridae) infecting F. asiaticum, the causal agent of wheat fusarium head blight. Phylogenetic analysis further suggested that FoMV3 is a new member of the genus Unuamitovirus within the family Mitoviridae. This is the first report of a new mitovirus associated with F. oxysporum f. sp. melonis., (© 2024. The Author(s), under exclusive licence to Springer-Verlag GmbH Austria, part of Springer Nature.)

Melon ( Cucumis melo L.) is a global commercial crop that is sensitive to seed-borne wilt infections caused by Fusarium oxysporum f. sp. melonis (Fom). To address the challenge of detecting Fom contamination, we designed a probe-based real-time PCR method, TDCP2, in combination with rapid or column-based DNA extraction protocols to develop reliable molecular detection methods. Utilizing TDCP2, the detection rate reached 100% for both artificially Fom-inoculated (0.25-25%) and pod-inoculated melon seeds in conjunction with DNA samples from either the rapid or column-based extraction protocol. We performed analyses of precision, recall, and F1 scores, achieving a maximum F1 score of 1 with TDCP2, which highlights the robustness of the method. Additionally, intraday and interday assays were performed, which revealed the high reproducibility and stability of column-based DNA extraction protocols combined with TDCP2. These metrics confirm the reliability of our developed protocols, setting a foundation for future enhancements in seed pathology diagnostics and potentially broadening their applicability across various Fom infection levels. In the future, we hope that these methods will reduce food loss by improving the control and management of melon diseases.

In September 2023, the UK Health Security Agency identified cases of Salmonella Saintpaul distributed across England, Scotland, and Wales, all with very low genetic diversity. Additional cases were identified in Portugal following an alert raised by the United Kingdom. Ninety-eight cases with a similar genetic sequence were identified, 93 in the United Kingdom and 5 in Portugal, of which 46% were aged under 10 years. Cases formed a phylogenetic cluster with a maximum distance of six single nucleotide polymorphisms (SNPs) and average of less than one SNP between isolates. An outbreak investigation was undertaken, including a case-control study. Among the 25 UK cases included in this study, 13 reported blood in stool and 5 were hospitalized. One hundred controls were recruited via a market research panel using frequency matching for age. Multivariable logistic regression analysis of food exposures in cases and controls identified a strong association with cantaloupe consumption (adjusted odds ratio: 14.22; 95% confidence interval: 2.83-71.43; p -value: 0.001). This outbreak, together with other recent national and international incidents, points to an increase in identifications of large outbreaks of Salmonella linked to melon consumption. We recommend detailed questioning and triangulation of information sources to delineate consumption of specific fruit varieties during Salmonella outbreaks.

The goal of this study was to characterize the bacterial diversity on different melon varieties grown in different regions of the US, and determine the influence that region, rind netting, and variety of melon has on the composition of the melon microbiome. Assessing the bacterial diversity of the microbiome on the melon rind can identify antagonistic and protagonistic bacteria for foodborne pathogens and spoilage organisms to improve melon safety, prolong shelf-life, and/or improve overall plant health. Bacterial community composition of melons (n = 603) grown in seven locations over a four-year period were used for 16S rRNA gene amplicon sequencing and analysis to identify bacterial diversity and constituents. Statistically significant differences in alpha diversity based on the rind netting and growing region (p < 0.01) were found among the melon samples. Principal Coordinate Analysis based on the Bray-Curtis dissimilarity distance matrix found that the melon bacterial communities clustered more by region rather than melon variety (R2 value: 0.09 & R2 value: 0.02 respectively). Taxonomic profiling among the growing regions found Enterobacteriaceae, Bacillaceae, Microbacteriaceae, and Pseudomonadaceae present on the different melon rinds at an abundance of ≥ 0.1%, but no specific core microbiome was found for netted melons. However, a core of Pseudomonadaceae, Bacillaceae, and Exiguobacteraceae were found for non-netted melons. The results of this study indicate that bacterial diversity is driven more by the region that the melons were grown in compared to rind netting or melon type. Establishing the foundation for regional differences could improve melon safety, shelf-life, and quality as well as the consumers' health., Competing Interests: The authors have declared that no competing interests exist., (Copyright: This is an open access article, free of all copyright, and may be freely reproduced, distributed, transmitted, modified, built upon, or otherwise used by anyone for any lawful purpose. The work is made available under the Creative Commons CC0 public domain dedication.)

Escherichia coli O157:H7 causes >73,000 foodborne illnesses in the United States annually, many of which have been associated with fresh ready-to-eat produce including cantaloupe melons. In this study, we created a produce-associated bacterial (PAB) library containing >7500 isolates and screened them for the ability to inhibit the growth of E. coli O157:H7 using an in vitro fluorescence-based growth assay. One isolate, identified by 16S and whole-genome sequence analysis as Enterobacter asburiae, was able to inhibit the growth of E. coli by ~30-fold in vitro and produced zones of inhibition between 13 and 21 mm against 12 E. coli outbreak strains in an agar spot assay. We demonstrated that E. asburiae AEB30 was able to grow, persist and inhibit the growth of E. coli on cantaloupe melons under simulated pre- and post-harvest conditions. Analysis of the E. asburiae AEB30 genome revealed an operon encoding a contact-dependent growth inhibition (CDI) system that when mutated resulted in the loss of E. coli growth inhibition. These data suggest that E. asburiae AEB30 is a potential biocontrol agent to prevent E. coli contamination of cantaloupe melons in both pre- and post-harvest environments and that its mode of action is via a CDI system., (Published 2024. This article is a U.S. Government work and is in the public domain in the USA. Microbial Biotechnology published by John Wiley & Sons Ltd.)

Aims: To identify biocontrol agents to prevent the growth of Salmonella serotype Enterica on cantaloupe melons during the pre- and postharvest periods., Methods and Results: We created a produce-associated bacterial library containing 8736 isolates and screened it using an in-vitro fluorescence inhibition assay to identify bacteria that inhibit the growth of S. Enterica. One isolate, Pantoea agglomerans ASB05, was able to grow, persist, and inhibit the growth of S. Enterica on intact cantaloupe melons under simulated pre- and postharvest conditions. We also demonstrated that the growth inhibition of S. Enterica by P. agglomerans ASB05 was due to the production of a phenazine type antibiotic., Conclusions: Pantoea agglomerans ASB05 is an effective biocontrol agent for the prevention of S. Enterica growth on intact cantaloupe melons in both the pre- and postharvest environments., (Published by Oxford University Press on behalf of Applied Microbiology International 2023.)

Ultrasound technology has been focused on due to its unique advantages in biofilm removal compared with traditional antibacterial methods. Herein, the anti-biofilm properties of low-frequency ultrasound (LFUS) were studied against Enterohemorrhagic Escherichia coli O157: H7 (E. coli O157:H7). After ultrasonication (20 kHz, 300 W) for 5 min, the removal rate of biofilm from polystyrene sheets reached up to 99.999 %. However, the bacterial cells could not be inactivated completely even extending the duration of ultrasonic irradiation to 30 min. Fortunately, this study indicated that LFUS could efficiently weaken the metabolic capacity and biofilm-forming ability of bacterial cells separated from biofilm. It could be associated with the removal of cell surface appendages and damage to cell membrane induced by mechanical vibration and acoustic cavitation. Besides, the genetic analysis proved that the transcription level of genes involved in curli formation was significantly down-regulated during ultrasonic irradiation, thus impeding the process of irreversible adhesion and cells aggregation. Finally, the actual application effect of LFUS was also evaluated in different fresh produces model. The results of this study would provide a theoretical basis for the further application of ultrasound in the food preservation., Competing Interests: Declaration of competing interest No conflict of interest exists in the submission of this paper., (Copyright © 2023 Elsevier B.V. All rights reserved.)

Key Message: Identified a recessive gene (Cmpmr2F) associated with resistance to infection by the powdery mildew causing agent Podosphaera xanthii race 2F. Powdery mildew (PM) is one of the most destructive fungal diseases of melon, which significantly reduces the crop yield and quality. Multiple studies are being performed for in-depth genetic understandings of PM-susceptibility or -resistance mechanisms in melon plants, but the holistic knowledge of the precise genetic basis of PM-resistance is unexplored. In this study, we characterized the recessive gene "Cmpmr2F" and found its association with resistance against the PM causative agent "Podosphaera xanthii race 2F." Fine genetic mapping revealed the major-effect region of a 26.25-kb interval on chromosome 12, which harbored the Cmpmr2F gene corresponding to the MELO3C002403, encoding allantoate amidohydrolase. The functional gene annotation, expression pattern, and sequence alignment analyses were carried out using two contrast parent lines of melon "X055" PM-susceptible and "PI 124112" PM-resistant. Further, gene silencing of Cmpmr2F using virus-induced gene silencing (VIGS) significantly increased PM-resistance in the susceptible plant. In contrast to the previously reported studies, we identified that Cmpmr2F-silenced plants showed no impairment in growth due to less apparent negative effects in silenced melon plants. So, it is believed that the Cmpmr2F gene has great potential for further breeding studies to increase the P. xanthii race 2F resistance in melon. In short, our study provides new genetic resources and a solid foundation for further functional analysis of PM-resistance genes in melon, as well as powerful molecular markers for marker-assisted breeding aimed at developing new melon varieties resistant to PM infection., (© 2023. The Author(s), under exclusive licence to Springer-Verlag GmbH Germany, part of Springer Nature.)

The genomic sequences segregating in experimental populations are often highly divergent from the community reference and from one another. Such divergence is problematic under various short-read-based genotyping strategies. In addition, large structural differences are often invisible despite being strong candidates for causal variation. These issues are exacerbated in specialty crop breeding programs with fewer, lower-quality sequence resources. Here, we examine the benefits of complete genomic information, based on long-read assemblies, in a biparental mapping experiment segregating at numerous disease resistance loci in the non-model crop, melon (Cucumis melo). We find that a graph-based approach, which uses both parental genomes, results in 19% more variants callable across the population and raw allele calls with a 2 to 3-fold error-rate reduction, even relative to single reference approaches using a parent genome. We show that structural variation has played a substantial role in shaping two Fusarium wilt resistance loci with known causal genes. We also report on the genetics of powdery mildew resistance, where copy number variation and local recombination suppression are directly interpretable via parental genome alignments. Benefits observed, even in this low-resolution biparental experiment, will inevitably be amplified in more complex populations., (© 2022. This is a U.S. Government work and not under copyright protection in the US; foreign copyright protection may apply.)

Nutrient disorder and presence of disease-causing agents in soilless media negatively influence the growth of muskmelon. To combat these issues, use of environmentally-friendly sanitation techniques is crucial for increased crop productivity. The study was conducted under greenhouse and field conditions to investigate the effect of two different sanitation techniques: steaming and formalin fumigation on various media's characteristics and their impact on muskmelon yield. Media: jantar, guar, wheat straw and rice hull and peat moss of 10% air-filled porosity and sanitized with formalin and steaming. Steaming of guar, jantar, and wheat straw increased the phosphorus (P) and potassium (K) concentrations by 13.80-14.86% and 6.22-8.45% over formalin fumigation. Likewise, P and K concentrations in muskmelon were higher under steaming. Steaming significantly inhibited the survival of Fusarium wilt sp. melonis, root knot nematode sp. meloidogyne and nitrifying bacteria in media than formalin fumigation. In conclusion, steaming decreased the prevalence of nitrifying bacteria and pathogens which thus improved the NO 3 - -N:NH 4 + -N ratios, P and K nutritional balance both in the media and muskmelon transplants. Hence, steaming as an environment-friendly approach is recommended for soilless media. Further, optimization of steaming for various composts with different crops needs to be investigated with steaming teachnique., (© 2021. The Author(s).)

Postharvest treatments with sanitizers and fungicides are applied to increase the quality, safety and shelf life of fresh produce including cantaloupes (also known as rockmelons). The primary role of sanitizers during cantaloupe washing is to prevent cross contamination of potentially pathogenic bacteria in washwater. Postharvest fungicide sprays or dips are employed to inhibit spoilage-causing fungi. While assessing the compatibility of these antimicrobials based on the measurement of active ingredients levels provides some indication of antimicrobial capacity, there is limited data on whether the interaction between these chemicals in wash water modifies their overall efficacy against relevant microorganisms. The aim of this research was to determine how chlorine- and peroxyacetic acid-based sanitizers interact with commercial guazatine- and imazalil-based fungicide formulations used on cantaloupes, and whether mixing these augments or suppresses anti-microbial activity against relevant human pathogens and spoilage fungi in wash water. The results were unpredictable: while most combinations were antimicrobial, the chlorine-based sanitizer when mixed with the guazatine-based fungicide had significantly reduced efficacy against pathogenic Salmonella spp. (~2.7 log) and the fungal spoilage organisms, Trichothecium roseum and Rhizopus stolonifera. Mixing the chlorine-based sanitizer with an imazalil-based fungicide produced a range of outcomes with antagonistic, indifferent and synergistic interactions observed for the fungal species tested. The peroxyacetic acid-based sanitizer led to indifferent interactions with the guazatine-based fungicide, while antagonism and synergy were observed when mixed with the imazalil-based fungicide. This study demonstrates that mixing postharvest agrichemicals used in the cantaloupe industry may increase the risk of microbial contamination and thereby potentially compromise food safety and quality., (Copyright © 2021. Published by Elsevier Ltd.)

Key Message: This is the first identification of QTLs underlying resistance to Pseudoperonospora cubensis in Cucumis melo using a genetically characterized isolate. Pseudoperonospora cubensis, causal organism of cucurbit downy mildew (CDM), is one of the largest threats to cucurbit production in the eastern USA. Currently, no Cucumis melo (melon) cultivars have significant levels of resistance. Additionally, little is understood about the genetic basis of resistance in C. melo. Recombinant inbred lines (RILs; N = 169) generated from a cross between the resistant melon breeding line MR-1 and susceptible cultivar Ananas Yok'neam were phenotyped for CDM resistance in both greenhouse and growth chamber studies. A high-density genetic linkage map with 5,663 binned SNPs created from the RIL population was utilized for QTL mapping. Nine QTLs, including two major QTLs, were associated with CDM resistance. Of the major QTLs, qPcub-10.1 was stable across growth chamber and greenhouse tests, whereas qPcub-8.2 was detected only in growth chamber tests. qPcub-10.1 co-located with an MLO-like protein coding gene, which has been shown to confer resistance to powdery mildew and Phytophthora in other plants. This is the first screening of C. melo germplasm with a genetically characterized P. cubensis isolate., (© 2021. The Author(s), under exclusive licence to Springer-Verlag GmbH Germany, part of Springer Nature.)

Cantaloupes contaminated with pathogens have led to many high-profile outbreaks and illnesses. Since bacterial virulence genes (VGs) can act in tandem with antibiotic-resistance and mobile genetic elements, there is a need to evaluate these gene reservoirs in fresh produce, such as cantaloupes. The goal of this study was to assess the distribution of antibiotic-resistance, virulence, and mobile genetic elements genes (MGEGs) in cantaloupe farm environments. A total of 200 samples from cantaloupe melons ( n  = 99), farm workers' hands ( n  = 66), and production water ( n  = 35) were collected in México. Each sample was assayed for the presence of 14 antibiotic-resistance genes, 15 VGs, and 5 MGEGs by polymerase chain reaction. Our results indicated that tetracycline ( tetA and tetB ) (18% of cantaloupe, 45% of hand samples) and sulfonamide ( sul1 ) (30% of cantaloupe, 71% of hand samples) resistance genes were frequently detected. The colistin resistance gene ( mcr1 ) was detected in 10% of cantaloupe and 23% of farm workers' hands. Among VGs, Salmonella genes invA and spiA were the most abundant. There was a significantly higher likelihood of detecting antibiotic-resistance, virulence, and MGEGs on hands compared with water samples. These results demonstrate a diverse pool of antibiotic-resistance and VGs in cantaloupe production.

Salmonella enterica is frequently implicated in foodborne disease outbreaks associated with fresh-cut fruits. In the U.S., more than one third of fruit-related outbreaks have been linked to two S. enterica serotypes Newport and Typhimurium. Approximately 80% of fruit-related human salmonellosis cases were associated with tomatoes, cantaloupes and cucumbers. In this study, we investigated the population dynamics of S. Newport and S. Typhimurium on fresh-cut tomato, cantaloupe, cucumber and apple under short-term storage conditions. We further compared the transcriptomic profiles of a S. Newport strain on fresh-cut tomato and cantaloupe using high-throughput RNA-seq. We demonstrated that both S. enterica Newport and Typhimurium survived well on various fresh-cut fruit items under refrigeration storage conditions, independent of inoculation levels. However, S. enterica displayed variable survival behaviors on different types of fruits. For example, at 7 d storage, the population of S. enterica reduced less than 0.2 log (p > 0.05) on fresh-cut tomato and cantaloupe, in contrast to ~0.5 log (p < 0.05) on cucumber and apple. RNA-seq analysis suggested that S. enterica mediates its survival on fresh-cut fruits through differentially regulating genes involved in specific carbon utilization and metabolic pathways. Several known bacterial virulence factors (e.g., pag gene) were found to be differentially regulated on fresh-cut tomato and cantaloupe, suggesting a link between the events of food contamination and subsequent human infection. Findings from this study contribute to a better understanding of S. enterica survival mechanisms on fresh-cut produce., (Copyright © 2021 Elsevier B.V. All rights reserved.)

In plants, chitin-triggered immunity is one of the first lines of defense against fungi, but phytopathogenic fungi have developed different strategies to prevent the recognition of chitin. Obligate biotrophs such as powdery mildew fungi suppress the activation of host responses; however, little is known about how these fungi subvert the immunity elicited by chitin. During epiphytic growth, the cucurbit powdery mildew fungus Podosphaera xanthii expresses a family of candidate effector genes comprising nine members with an unknown function. In this work, we examine the role of these candidates in the infection of melon (Cucumis melo L.) plants, using gene expression analysis, RNAi silencing assays, protein modeling and protein-ligand predictions, enzymatic assays, and protein localization studies. Our results show that these proteins are chitinases that are released at pathogen penetration sites to break down immunogenic chitin oligomers, thus preventing the activation of chitin-triggered immunity. In addition, these effectors, designated effectors with chitinase activity (EWCAs), are widely distributed in pathogenic fungi. Our findings reveal a mechanism by which fungi suppress plant immunity and reinforce the idea that preventing the perception of chitin by the host is mandatory for survival and development of fungi in plant environments., (� American Society of Plant Biologists 2021. All rights reserved. For permissions, please email: journals.permissions@oup.com.)

Inoculation studies are important when assessing microbial survival and growth in food products. These studies typically involve the pregrowth of multiple strains of a target pathogen under a single condition; this emphasizes strain diversity. To gain a better understanding of the impacts of strain diversity ("nature") and pregrowth conditions ("nurture") on subsequent bacterial growth in foods, we assessed the growth and survival of Salmonella enterica ( n  = 5), Escherichia coli ( n  = 6), and Listeria ( n  = 5) inoculated onto tomatoes, precut lettuce, and cantaloupe rind, respectively. Pregrowth conditions included (i) 37°C to stationary phase (baseline), (ii) low pH, (iii) high salt, (iv) reduced water activity, (v) log phase, (vi) minimal medium, and (vii) 21°C. Inoculated tomatoes were incubated at 21°C; lettuce and cantaloupe were incubated at 7°C. Bacterial counts were assessed over three phases, including initial reduction (phase 1), change in bacterial numbers over the first 24 h of incubation (phase 2), and change over the 7-day incubation (phase 3). E. coli showed overall decline in counts (<1 log) over the 7-day period, except for a <1-log increase after pregrowth in high salt and to mid-log phase. In contrast, S. enterica and Listeria showed regrowth after an initial reduction. Pregrowth conditions had a substantial and significant effect on all three phases of S. enterica and E. coli population dynamics on inoculated produce, whereas strain did not show a significant effect. For Listeria , both pregrowth conditions and strain affected changes in phase 2 but not phases 1 and 3. IMPORTANCE Our findings suggest that inclusion of multiple pregrowth conditions in inoculation studies can best capture the range of growth and survival patterns expected for Salmonella enterica and Escherichia coli present on produce. This is particularly important for fresh and fresh-cut produce, where stress conditions encountered by pathogens prior to contamination can vary widely, making selection of a typical pregrowth condition virtually impossible. Pathogen growth and survival data generated using multiple pregrowth conditions will allow for more robust microbial risk assessments that account more accurately for uncertainty., (Copyright © 2021 American Society for Microbiology.)

Cantaloupes have emerged as significant vehicles of widespread foodborne illness outbreaks caused by bacterial pathogens, including Salmonella. The purpose of this study was to investigate the efficiency of Salmonella colonization and internalization in cantaloupes by relevant routes of contamination. Cantaloupe plants (Cucumis melo 'reticulatus') from two cultivars 'Athena' (Eastern) and 'Primo' (Western) were grown from commercial seed. Plants were maintained in the NCSU BSL-3P phytotron greenhouse. Salmonella enterica (a cocktail of cantaloupe-associated outbreak serovars Javiana, Newport, Panama, Poona and Typhimurium) contamination was introduced via blossoms or soil at ca. 4.4 log 10  CFU/blossom or 8.4 log 10  CFU/root zone, respectively. Cantaloupes were analyzed for Salmonella by enrichment in accordance with modified FDA-BAM methods. Five randomly chosen colonies from each Salmonella-positive sample were typed using the Agilent 2100 bioanalyzer following multiplex PCR. Data were analyzed for prevalence of contamination and serovar predominance in fruit, stems and soil. Of the total cantaloupe fruit harvested from Salmonella-inoculated blossoms (n = 63), 89% (56/63) were externally contaminated and 73% (46/63) had Salmonella internalized into the fruit. Serovar Panama was the most commonly isolated from the surface of fruit while S. Panama and S. Poona were the most prevalent inside the fruit. When soil was inoculated with Salmonella at one day post-transplant, 13% (8/60) of the plants were shown to translocate the organism to the lower stem (ca. 4 cm) by 7 days post-inoculation (dpi). We observed Salmonella persistence in the soil up to 60 dpi with S. Newport being the predominant serovar at 10 and 20 dpi. These data demonstrate that contaminated soil and blossoms can lead to Salmonella internalization into the plant or fruit at a relatively high frequency., (Published by Elsevier Ltd.)

Cadmium (Cd), prevailing in most of the agricultural lands of the world contaminates food chain, thereby causing several health implications. It has become the main heavy metal contaminant in most of the agricultural lands of Pakistan due to the widespread use of phosphate fertilizers besides application of irrigation water contaminated with industrial and mining effluents. Plant growth promoting bacteria (PGPB) are capable to enhance growth and metal stress tolerance in supplemented plants. Zinc oxide nanoparticles (ZnO-NPs) are capable to alleviate various abiotic stresses when applied to plants. During current research, the efficacy of single and combined application of Bacillus fortis IAGS 223 and ZnO-NPs was evaluated for alleviation of Cd (75 mg kg -1 ) induced phytotoxicity in Cucumis melo plants. For this purpose, C. melo plants, subjected to Cd stress were treated with B. fortis IAGS 223 and ZnO-NPs (20 mg kg -1 ), either alone or in combination. The growth relevant characteristics including photosynthetic pigments, hydrogen peroxide (H 2 O 2 ), malondialdehyde (MDA), and activities of antioxidative enzymes as well as Zn and Cd contents in treated plants were examined. The individual application of ZnO-NPs and B. fortis IAGS 223 slightly enhanced all the above-mentioned growth characteristics in plants under Cd stress. However, the combined application of ZnO-NPs and B. fortis IAGS-223 considerably modulated the activity of antioxidant enzymes besides upgradation of the biochemicals and growth parameters of Cd stressed plants. The decreased amount of stress markers such as H 2 O 2 , and MDA in addition with reduction of Cd contents was observed in shoots of ZnO-NPs and B. fortis IAGS-223 applied plants. B. fortis IAGS-223 inoculated plants supplemented with ZnO-NPs, exhibited reduced amount of Cd as well as protein bound thiols and non-protein bound thiols under Cd stress. Subsequently, the reduced Cd uptake improved growth of ZnO-NPs and B. fortis IAGS-223 applied plants. Henceforth, field trials may be performed to formulate appropriate combination of ZnO-NPs and B. fortis IAGS-223 to acquire sustainable crop production under Cd stress., (Copyright © 2020 Elsevier Masson SAS. All rights reserved.)

Salmonella enterica serovar Typhimurium is a pathogen harbored by livestock and shed in their feces, which serves as an acquisition source for adult house flies. This study used a green fluorescent protein (GFP) expressing strain of Salmonella Typhimurium to assess its acquisition by and survival within house flies, and transmission from and between flies in the presence or absence of cantaloupe. Female house flies were exposed to manure inoculated with either sterile phosphate-buffered saline or GFP- Salmonella Typhimurium for 12 h, then used in four experiments each performed over 24 h. Experiment 1 assessed the survival of GFP- Salmonella Typhimurium within inoculated flies. Experiment 2 determined transmission of GFP- Salmonella Typhimurium from inoculated flies to cantaloupe. Experiment 3 assessed fly acquisition of GFP- Salmonella Typhimurium from inoculated cantaloupe. Experiment 4 evaluated transmission of GFP- Salmonella Typhimurium between inoculated flies and uninoculated flies in the presence and absence of cantaloupe. GFP- Salmonella Typhimurium survived in inoculated flies but bacterial abundance decreased between 0 and 6 h without cantaloupe present and between 0 and 6 h and 6 and 24 h with cantaloupe present. Uninoculated flies acquired GFP- Salmonella Typhimurium from inoculated cantaloupe and bacterial abundance increased in cantaloupe and flies from 6 to 24 h. More uninoculated flies exposed to inoculated flies acquired GFP- Salmonella Typhimurium when cantaloupe was present than when absent. We infer that the presence of a shared food source facilitated the transfer of GFP- Salmonella Typhimurium from inoculated to uninoculated flies. Our study demonstrated that house flies acquired, harbored, and excreted viable GFP- Salmonella Typhimurium and transferred bacteria to food and each other. Understanding the dynamics of bacterial acquisition and transmission of bacteria between flies and food helps in assessing the risk flies pose to food safety and human health.

Listeria monocytogenes is a foodborne pathogen that causes high rates of hospitalization and mortality in people infected. Contamination of fresh, ready to eat produce by this pathogen is especially troubling because of the ability of this bacterium to grow on produce under refrigeration temperatures. In this study, we created a library of over 8,000 plant phyllosphere-associated bacteria and screened them for the ability to inhibit the growth of L. monocytogenes in an in vitro fluorescence-based assay. One isolate, later identified as Bacillus amyloliquefaciens ALB65, was able to inhibit the fluorescence of L. monocytogenes by >30-fold in vitro. B. amyloliquefaciens ALB65 was also able to grow, persist, and reduce the growth of L. monocytogenes by >1.5 log CFU on cantaloupe melon rinds inoculated with 5 × 10 3 CFU at 30°C and was able to completely inhibit its growth at temperatures below 8°C. DNA sequence analysis of the B. amyloliquefaciens ALB65 genome revealed six gene clusters that are predicted to encode genes for antibiotic production; however, no plant or human virulence factors were identified. These data suggest that B. amyloliquefaciens ALB65 is an effective and safe biological control agent for the reduction of L. monocytogenes growth on intact cantaloupe melons and possibly other types of produce. IMPORTANCE Listeria monocytogenes is estimated by the Centers for Disease Control and Prevention and the U.S. Food and Drug Administration to cause disease in approximately 1,600 to 2,500 people in the United States every year. The largest known outbreak of listeriosis in the United States was associated with intact cantaloupe melons in 2011, resulting in 147 hospitalizations and 33 deaths. In this study, we demonstrated that Bacillus amyloliquefaciens ALB65 is an effective biological control agent for the reduction of L. monocytogenes growth on intact cantaloupe melons under both pre- and postharvest conditions. Furthermore, we demonstrated that B. amyloliquefaciens ALB65 can completely inhibit the growth of L. monocytogenes during cold storage (<8°C)., (Copyright © 2020 American Society for Microbiology.)

This study evaluated the efficacy of combined chitosan (Chi) and Cymbopogon citratus (DC) Stapf. essential oil (CCEO) to inhibit the fungal phytopathogen Paramyrothecium roridum L. Lombard & Crous and control crater rot in melon (Cucumis melo L.). Effects of several Chi and CCEO concentrations to inhibit the growth of four P. roridum isolates in vitro, as well as the type of interaction of some combined concentrations of Chi and CCEO was evaluated. Effects of coatings with combined concentrations of Chi and CCEO on development of crater rot lesions in melon artificially inoculated with P. roridum during storage (15 days, 25 °C) were measured. Chi (2.5, 3.75, 5, and 6.75 mg/mL) and CCEO (0.3 and 0.6 μL/mL) led to growth inhibition of the four examined P. roridum isolates. Combinations of Chi (5 mg/mL) and CCEO (0.15 and 0.3 μL/mL) had additive interaction to inhibit P. roridum. Coatings with additive combined concentrations of Chi and CCEO decreased the development and severity of carter rot lesions in melon during room storage regardless of the inoculated P. roridum isolate. Therefore, application of coatings formulated with combined concentrations of Chi and CCEO could be alternative strategies to control crater rot caused by P. roridum in melon and decrease synthetic fungicide use in this fruit.

Foodborne pathogens constitute a major food safety risk for cantaloupe, and pathogen biofilms formed are particularly difficult to remove. The goal of this study was to evaluate abrasive brushing in removing biofilms from cantaloupe surface using Listeria monocytogenes V7 and Salmonella enterica serovar Typhimurium ATCC 13311 as models. Cantaloupe rind pieces were inoculated and then subjected to 30 min or 24 h incubations. The incubated rind pieces were subject to different washing and/or brushing procedures and then the bacterial cells were enumerated. Cryo-SEM showed planktonic cells after 30 min incubation, whereas biofilms formed at rind surfaces after 24 h incubation. For L. monocytogenes 30 min incubation group, the log reductions were 1.4 for brushing, 2.1 for brushing with diatomaceous earth (DE), 2.8 for peroxyacetic acid (PAA) washing, 4.2 for brushing with PAA, and 4.0 for brushing with DE and PAA. In contrast, for the 24 h incubation group, the log reductions were 0.4 for brushing, 1.5 for brushing with DE, 1.1 for PAA washing, 1.6 for brushing with PAA, and 3.0 for brushing with DE and PAA. Similar results were observed for the S. Typhimurium group. These outcomes showed the pathogen-removal efficacy of brushing with DE (abrasive brushing) in the presence of biofilms, suggesting the potential application of abrasive brushing in cleaning cantaloupe for improved food safety., (Copyright © 2020 Elsevier B.V. All rights reserved.)

The common use of gloves in retail practices represents a potential route for cross contamination of foodborne pathogens in fresh-cut produce. Using fresh-cut cantaloupe as a food model, we investigated factors that may influence glove-mediated cross contamination by Listeria monocytogenes and developed mathematical models to illustrate the patterns of transfer during fresh-cutting practices. Contact time (2, 5, 10 s), contact pressure (0.05, 0.18, 0.37 psi), and glove type (nitrile, polyvinyl chloride, polyethylene) did not have a significant effect on transfer of L. monocytogenes from cantaloupe rind to flesh, or from flesh to flesh. However, glove type appeared to affect L. monocytogenes transfer from the stem scar tissue to cantaloupe flesh (P = 0.0371). Transfer from rind pieces that had been washed with water was significantly higher than transfer from pieces that had not been washed (P = 0.0006). Predictive modeling and experimental validation suggested that transfer of L. monocytogenes on cantaloupe flesh persists over 85 pieces through consecutive contacts with a gloved hand. Findings of the study provide new scientific data to aid researchers, retailers, and caterers in safety risk assessments of fresh-cut practices used to prepare cantaloupes and other produce items., (Copyright © 2019 Elsevier Ltd. All rights reserved.)

Various produce including cantaloupe, caramel-coated apples, and packaged salads, have been recognized in recent years as vehicles for listeriosis, a human foodborne disease caused by intracellular pathogen Listeria monocytogenes. Our knowledge regarding the role of these foods in L. monocytogenes virulence, however, is limited. Understanding their role in modulating L. monocytogenes virulence can be useful in risk assessments and for developing control measures. In this study, we employed the Galleria mellonella larvae model to evaluate virulence potential of fifteen clinical, environmental and food isolates of L. monocytogenes, related to three major outbreaks, after growth on different foods. The non-human pathogen Listeria innocua was also included in the panel. Strains were inoculated in parallel in 5ml of brain heart infusion (BHI) broth, and on the surfaces of cantaloupe and apple fragments (5g each) at about 105 colony forming units (CFU)/ml/fragment. One set of inoculated broth and food fragments was incubated at 10°C for 5 days while the second set was kept at 25°C for 3 days. L. monocytogenes cells were recovered from the fruits and BHI, washed twice, re-suspended in saline, and used to inoculate G. mellonella larvae at final concentrations of 106 and 105 CFU/larva. The larvae were incubated at 37°C and monitored for mortality (LT50-time taken to kill 50% of the larvae) and phenotypic changes over seven days. L. monocytogenes grown on cantaloupe and apple flesh surfaces resulted in higher virulence than when grown in BHI. L. monocytogenes infection at 106 CFU/larvae resulted in an average LT50 of ≤ 30, 36 and 47 hours on cantaloupe, apples and BHI, respectively. These results represent a 2.5-4-fold increased mortality compared with an LT50 ≥120 hours in larvae infected with the same doses of L. innocua grown in corresponding matrices. Similar trends were also recorded with doses of about 105 CFU /larvae., Competing Interests: The authors have declared that no competing interests exist.

In the present study, we investigated whether cold plasma activation affected the efficacy of aerosolized hydrogen peroxide against S. Typhimurium and L. innocua. Stem scars and smooth surfaces of grape tomatoes, surfaces of Granny Smith apples and Romaine lettuce (both midrib and upper leaves) and cantaloupe rinds were inoculated with two-strain cocktails of S. Typhimurium and 3-strain cocktails of L. innocua. The inoculated samples were treated with 7.8% aerosolized H 2 O 2 with and without cold plasma for various times. For all fresh produce items and surfaces, cold plasma significantly (P < 0.05) improved the efficacy of aerosolized H 2 O 2 against Salmonella and L. innocua. Without cold plasma activation, H 2 O 2 aerosols only reduced populations of Salmonella by 1.54-3.17 log CFU/piece while H 2 O 2 with cold plasma achieved 2.35-5.50 log CFU/piece reductions of Salmonella. L. innocua was more sensitive to the cold plasma-activated H 2 O 2 than Salmonella. Cold plasma activated H 2 O 2 aerosols reduced Listeria populations by more than 5 log CFU/piece on all types and surfaces of fresh produce except for the tomato stem scar area. Without cold plasma, the reductions by H 2 O 2 were only 1.35-3.77 log CFU/piece. Overall, our results demonstrated that cold plasma activation significantly enhanced the efficacy of H 2 O 2 mist against bacteria on fresh produce., (Published by Elsevier Ltd.)

Bacterial fruit blotch (BFB) causes losses in melon marketable yield. However, until now, there has been no information about the genetic loci responsible for resistance to the disease or their pattern of inheritance. We determined the inheritance pattern of BFB resistance from a segregating population of 491 F 2 individuals raised by crossing BFB-resistant (PI 353814) and susceptible (PI 614596) parental accessions. All F 1 plants were resistant to Acidovorax citrulli strain KACC18782, and F 2 plants segregated with a 3:1 ratio for resistant and susceptible phenotypes, respectively, in a seedling bioassay experiment, indicating that BFB resistance is controlled by a monogenic dominant gene. In an investigation of 57 putative disease-resistance related genes across the melon genome, only the MELO3C022157 gene (encoding TIR-NBS-LRR domain), showing polymorphism between resistant and susceptible parents, revealed as a good candidate for further investigation. Cloning, sequencing and quantitative RT-PCR expression of the polymorphic gene MELO3C022157 located on chromosome 9 revealed multiple insertion/deletions (InDels) and single nucleotide polymorphisms (SNPs), of which the SNP A 2035 T in the second exon of the gene caused loss of the LRR domain and truncated protein in the susceptible accession. The InDel marker MB157-2, based on the large (504 bp) insertion in the first intron of the susceptible accession, was able to distinguish resistant and susceptible accessions among 491 F 2 and 22 landraces/inbred accessions with 98.17% and 100% detection accuracy, respectively. This novel PCR-based, co-dominant InDel marker represents a practical tool for marker-assisted breeding aimed at developing BFB-resistant melon accessions., Competing Interests: The authors declare that they have no conflicts of interest.

Background and Objective: Fresh-cut cantaloupe faces several quality challenges as an increase of decay rate, browning, respiration and other changes which can reduce the shelf life. Nano-coating combination films as chitosan-nano-silicon dioxide and chitosan-nano-titanium dioxide can extend the shelf life of cantaloupe pieces., Materials and Methods: Cantaloupes were classified into four sets, Control (C), Chitosan (CTSN), Chitosan-Nano-Silicon Dioxides (CTSN/NSD) and Chitosan-Nano-titanium Dioxide (CTSN/NTD)., Results: Coated cantaloupe pieces had the best browning activity of polyphenol oxidase (PPO) (0.26 U/min/g) and the lowest water activity (Aw) (0.89) by chitosan-nano-silicon dioxide films. Chitosan-nano-titanium dioxide films could maintain weight loss (0.93%) and color pigments for L* value. It worked effectively against lipid peroxidation (MDA) (0.15 nmol g-1), Peroxidase Activity (POD) (3.92 U/min/g) and microbial contaminations (6.44 log CFU g-1)., Conclusion: Both nano-coating films improved some chemical parameters as Titratable Acidity (TA), Soluble Solid Content (SSC) and pH. Nano-coatings films techniques were recommended for cantaloupes preservation methods.

Listeria monocytogenes is an intracellular pathogen responsible for listeriosis, a foodborne disease that can lead to life-threatening meningitis. The 2011 L. monocytogenes cantaloupe outbreak was among the deadliest foodborne outbreaks in the United States. We conducted in vitro and in vivo infection analyses to determine whether strains LS741 and LS743, two clinical isolates from the cantaloupe outbreak, differ significantly from the common laboratory strain 10403S. We showed that LS741 and LS743 exhibited increased virulence, characterized by higher colonization of the brain and other organs in mice. Assessment of cellular immune responses to known CD8 + T cell antigens was comparable between all strains. However, pre-existing immunity to 10403S did not confer protection in the brain against challenge with LS741. These studies provide insights into the pathogenesis of clinical isolates linked to the 2011 cantaloupe outbreak and also indicate that currently utilized laboratory strains are imperfect models for studying L. monocytogenes pathogenesis.

Foodborne illness associated with fresh, ready-to-eat produce continues to be a significant challenge to public health. In this study, we created a phyllosphere-associated lactic acid bacteria (PLAB) library and screened it via a high-throughput in vitro fluorescent assay to identify bacteria capable of inhibiting the growth of the pathogenic bacterium Salmonella enterica . One isolate, 14B4, inhibited the growth of S. enterica by >45-fold in vitro; it was able to grow and persist on the surfaces of cantaloupe melons at both ambient (25°C) and refrigerator (5°C) temperatures. Isolate 14B4 inhibited the growth of S. enterica on the surfaces of cantaloupes by >3 log when incubated at 25°C for 24 h and by >4 log when the cantaloupes were stored at 5°C for 3 days and the temperature was shifted to 25°C for 2 days. Genomic DNA sequence analysis of isolate 14B4 revealed that it was Lactococcus lactis and that it did not contain any known antibiotic biosynthesis gene clusters, antibiotic resistance genes, or genes encoding any known virulence factors. Organic acid analysis revealed that L. lactis produces substantial amounts of lactic acid, which is likely the inhibitory substance that reduced the growth of Salmonella on the cantaloupes.

Listeria monocytogenes, the causative agent of listeriosis in humans, is a Gram-positive bacterium that is contracted via the ingestion of contaminated foods. Two of the largest outbreaks of listeriosis occurred following consumption of tainted cantaloupe and packaged salads. Molecular methods and immuno-based techniques for detection of L. monocytogenes in these food matrices can be difficult due to the presence of assay inhibiting elements. In this study, we utilized a novel enrichment media containing activated charcoal as the key ingredient that induces hyperactive expression and secretion of L. monocytogenes virulence proteins. The Bio-Plex suspension array system, based on Luminex xMAP technology, was subsequently employed to specifically detect accumulated L. monocytogenes secreted and membrane bound proteins via paramagnetic microsphere-antibody complexes. Cantaloupe and packaged salad samples were treated with a dilution series of L. monocytogenes and incubated in activated charcoal media following a short pre-enrichment step in Buffered Listeria Enrichment Broth. Secreted L. monocytogenes lysteriolysin O was captured using magnetic microsphere-antibody conjugates and measured using the Bio-Ple×200 analyzer. As few as 10 0  CFU/g of L. monocytogenes was detected from both spiked cantaloupe and packaged salad samples. In addition, antibody conjugated microspheres targeting a membrane protein present on both pathogenic and nonpathogenic Listeria species was used to identify as few as 10 0  CFU/g of both pathogenic and nonpathogenic species in cantaloupe and packaged salad. This method presumptively identifies L. monocytogenes from cantaloupe and packaged salad in less than 24 h and non-pathogenic Listeria species within 22 h., (Copyright © 2019. Published by Elsevier Ltd.)

The purpose of this study was to evaluate the effectiveness of the combination treatment of lactic acid immersion and saturated or superheated steam (SHS) on inactivation of foodborne pathogens on cantaloupes. Saturated steam (SS) treatments were performed at 100 °C, while SHS treatments were delivered at either 150 or 200 °C. Escherichia coli O157:H7, Salmonella Typhimurium and Listeria monocytogenes-inoculated cantaloupes were exposed to 2% lactic acid or sterile distilled water for 1 min followed by a maximum of 20 s of SS or SHS. Populations of each of the three pathogens on cantaloupes were reduced to under the detection limit (1.0 log CFU/cm 2 ) after the combination treatment of 2% lactic acid and 200 °C steam for 20 s. To compare the effect of the lactic acid treatment method, we conducted spray application with 2% lactic acid combined with SS or SHS treatment; however, no significant log reduction differences were found between immersion and spraying techniques. After combination treatment of cantaloupes for 20 s, color and maximum load values (a characteristic of texture) were not significantly different from those of untreated controls. The results of this study suggest that the combination treatment of lactic acid and SHS can be used as an antimicrobial intervention for cantaloupes without inducing quality deterioration., (Copyright © 2019. Published by Elsevier Ltd.)

The fungus causing target spot disease, Corynespora cassiicola (Berk. & M. A. Curtis) C. T. Wei, poses an increasing threat to watermelon (Citrullus lanatus), muskmelon (Cucumis melo), and cucumber (Cucumis sativus); the most economically important cucurbit crops grown in China. An understanding of the molecular mechanisms underlying the pathogenicity of C. cassiicola is essential for the development of new strategies to control this disease-causing fungus. Agrobacterium tumefaciens-mediated transformation (ATMT) might be useful to obtain transformants of C. cassiicola, for the ultimate identification of genes involved in pathogenicity. In the present work, we established and optimized an ATMT protocol using A. tumefaciens strain AGL-1 carrying the vector pATMT1 for C. cassiicola. Efficiency of ATMT was 102-148 transformants per 10 6 conidia and successive subculturing of transformants on non-selective and selective media demonstrated that the integrated transfer (T)-DNA was stably inherited in C. cassiicola transformants. The integration of the hygromycin B phosphotransferase (hph) gene into C. cassiicola was validated by PCR and Southern blot analyses, which revealed that nearly 90 % of the transformants contained single-copy T-DNA. The transformants with altered phenotypes were characterized. Three of these transformants completely lost pathogenicity and other three showed strongly impaired pathogenicity relative to the Cc-GX strain on muskmelon leaves. These results strongly suggest that ATMT may be used as a molecular tool for identifying genes relevant to pathogenicity in the fungus C. cassiicola, an emerging threat to several agronomically important plants in China., (Copyright © 2019 The Authors. Published by Elsevier Ltd.. All rights reserved.)

Background: Cantaloupe melon (Cucumis melo L.) is one of the most important dessert fruits and is cultivated in many countries of the world. The effects of microwave (400 and 800 W for 110 s), ohmic (100 and 200 V for 110 s) and conventional heating (27-75 °C for 30 min) treatments on inactivation of Escherichia coli, Salmonella Typhimurium, S. Enteritidis and Staphylococcus aureus pathogens; pH and degradation of vitamin C, β-carotene and phenolic compounds of cantaloupe juice were investigated., Results: As time passed, all of the treatments resulted in significant (P ≤ 0.05) decreases in the number of pathogens and vitamin C, β-carotene and phenolic compound content, whereas the pH of samples did not show significant changes. The mentioned parameters were more reduced by increasing the power, voltage and temperature of ohmic, microwave and conventional heating treatments, respectively. Comparison of the results for conventional heating with those of ohmic and microwave heating revealed that the complete inactivation time of pathogens by the two latter treatments was much shorter than that of the former. After 20 s, the effect of ohmic heating at 200 V on decreasing vitamin C content was significantly (P ≤ 0.05) higher than that of the other treatments. The amounts of β-carotene and phenolic compounds showed the most reduction under 800 W microwave treatment., Conclusion: The results obtained for conventional, ohmic and microwave heating treatments indicated a higher degradation of β-carotene and phenolic compounds and a lower loss of vitamin C in the former. © 2019 Society of Chemical Industry., (© 2019 Society of Chemical Industry.)

Substrates are fundamental prerequisites for growing grafted seedlings. In this study, substrates with different pH levels (5.0, 5.5, 6.0, 6.5, 7.0, and 8.0) were set up to elucidate the effect of pH on cadmium (Cd) uptake in grafted muskmelon (Cucumis melo L.) plants. Bacterial diversity was also investigated. Results showed that pH and high Cd concentration greatly affected the growth of grafted plants. The chlorophyll content of the muskmelon leaves decreased at 100 μM Cd. The majority of the Cd ions accumulated in the rootstock rather than in the shoot tissue in all of the treatments. The shoots and roots showed the highest Cd content at pH 5.5 and the lowest Cd content at pH 8.0 regardless of the Cd concentration. The operational taxonomic units belonging to Proteobacteria and Bacteroidetes were significantly (p < 0.05) enriched at different substrate pH levels compared with those at pH 5.0. The operational taxonomic units belonging to the phyla Firmicutes, Acidobacteria, and Chloroflexi were significantly decreased. The available nitrogen, phosphorus, Cd, and pH were strongly linked to bacterial community compositions. On the contrary, the available potassium was weakly correlated with the bacterial structure. This study demonstrates that pH greatly affects Cd uptake in grafted muskmelon plants and predicts microbial community structures in breeding substrates with different pH levels. Our results suggest that Cd accumulation in grafted plants can be reduced by setting the appropriate substrate pH. This work can serve as a reference for growing high-quality grafted plants and ensuring food safety in the presence of Cd contamination., (Copyright © 2019 Elsevier Masson SAS. All rights reserved.)

The cucurbit powdery mildew elicited by Podosphaera xanthii is one of the most important limiting factors in cucurbit production. Our knowledge of the genetic and molecular bases underlying the physiological processes governing this disease is very limited. We used RNA-sequencing to identify differentially expressed genes in leaves of Cucumis melo upon inoculation with P. xanthii, using RNA samples obtained at different time points during the early stages of infection and their corresponding uninfected controls. In parallel, melon plants were phenotypically characterized using imaging techniques. We found a high number of differentially expressed genes (DEGs) in infected plants, which allowed for the identification of many plant processes that were dysregulated by the infection. Among those, genes involved in photosynthesis and related processes were found to be upregulated, whereas genes involved in secondary metabolism pathways, such as phenylpropanoid biosynthesis, were downregulated. These changes in gene expression could be functionally validated by chlorophyll fluorescence imaging and blue-green fluorescence imaging analyses, which corroborated the alterations in photosynthetic activity and the suppression of phenolic compound biosynthesis. The powdery mildew disease in melon is a consequence of a complex and multifaceted process that involves the dysregulation of many plant pathways such as primary and secondary metabolism.

Mitochondria play an essential part in fighting against pathogen infection in the defense responses of fruits. In this study, we investigated the reactive oxygen species (ROS) production, energy metabolism, and changes of mitochondrial proteins in harvested muskmelon fruits ( Cucumis melo cv. Yujinxiang) inoculated with Trichothecium roseum. The results indicated that the fungal infection obviously induced the H 2 O 2 accumulation in mitochondria. Enzyme activities were inhibited in the first 6 h postinoculation (hpi), including succinic dehydrogenase, cytochrome c oxidase, H + -ATPase, and Ca 2+ -ATPase. However, the activities of Ca 2+ -ATPase and H + -ATPase and the contents of intracellular adenosine triphosphate (ATP) were improved to a higher level at 12 hpi. A total of 42 differentially expressed proteins were identified through tandem mass tags-based proteomic analyses, which are mainly involved in energy metabolism, stress responses and redox homeostasis, glycolysis and tricarboxylic acid cycle, and transporter and mitochondria dysfunction. Taken together, our results suggest that mitochondria play crucial roles in the early defense responses of muskmelons against T. roseum infection through regulation of ROS production and energy metabolism.

Several outbreaks of listeriosis have implicated fresh produce but genetic factors required for growth of Listeria monocytogenes on produce remain poorly characterized. Based on the fact that β-lactam antibiotics only kill bacterial cells that are growing, we hypothesized that ampicillin selection can enrich for L. monocytogenes mutants unable to grow on produce. For validation, we examined relative recovery of L. monocytogenes strain 2011L-2858 and its cold-sensitive mutant L1E4 following inoculation of cantaloupe rind fragments with 1:1 mixture of the strains and incubation at 4°C with or without ampicillin. Listeria monocytogenes from rind fragments inoculated with the mixed cultures and incubated in the presence of ampicillin were used to inoculate fresh rind fragments for a second round of enrichment. In the presence of ampicillin, the proportion of L1E4 increased from 55% on day 0 to 78% on day 14, with higher recovery (85% after 14 days) in the second round of enrichment. These data suggested that L1E4 was enriched on cantaloupe rind fragments while growing cells of the wildtype were killed by ampicillin. Application of this protocol to transposon mutant libraries from three L. monocytogenes strains yielded several mutants unable to grow on cantaloupe. Thus, ampicillin selection can facilitate discovery of genes essential for growth of L. monocytogenes on fresh produce., (© FEMS 2019.)

Feeding of sucking insects, such as the rice brown planthopper (Nilaparvata lugens; BPH), causes only limited mechanical damage on plants that is otherwise essential for injury-triggered defense responses against herbivores. In pursuit of complementary BPH elicitors perceived by plants, we examined the potential effects of BPH honeydew secretions on the BPH monocot host, rice (Oryza sativa). We found that BPH honeydew strongly elicits direct and putative indirect defenses in rice, namely accumulation of phytoalexins in the leaves, and release of volatile organic compounds from the leaves that serve to attract natural enemies of herbivores, respectively. We then examined the elicitor active components in the honeydew and found that bacteria in the secretions are responsible for the activation of plant defense. Corroborating the importance of honeydew-associated microbiota for induced plant resistance, BPHs partially devoid of their microbiota via prolonged antibiotics ingestion induced significantly less defense in rice relative to antibiotic-free insects applied to similar groups of plants. Our data suggest that rice plants may additionally perceive herbivores via their honeydew-associated microbes, allowing them to discriminate between incompatible herbivores-that do not produce honeydew-and those that are compatible and therefore dangerous., (© The Author(s) 2019. Published by Oxford University Press on behalf of the Society for Experimental Biology.)

The most commonly used indicator of fecal contamination in fresh produce production and packing is Escherichia coli. In depth analysis of the prevalence and characteristics of naturally occurring E. coli strains in these environments is important because it can (1) serve as an indicator of sources of fecal contamination; and (2) provide information on strain pathogenicity, persistence, and other defining characteristics such as multidrug resistance. In this study, we analyzed 341 E. coli strains isolated from the jalapeño pepper, tomato and cantaloupe farm environments, in Northeast Mexico. Strains were isolated from produce, farmworkers' hands, soil and water. Pathotypes, genotypes, biofilm formation and antibiotic resistance were characterized. Phylogenetic subgroups and identification of diarrheagenic E. coli were determined by PCR; biofilm formation was quantified using a plate-based colorimetric method. Antibiotic resistance was analyzed by the Kirby Bauer diffusion disc method. Most isolates (N = 293, 86%) belonged to phylogenetic group A. Only four isolates (1.2%) were diarrheagenic: EPEC (N = 3) and ETEC (N = 1). Antibiotic resistance to tetracycline (23.2%) and ampicillin (19.9%) was high, and only 3.5% of the strains presented resistance to >5 antibiotics. Biofilms were produced by most strains (76%), among which 34.4% were categorized as high producers. The presence of antibiotic resistant E. coli strains that may contain gene markers for pathogenicity and which can form biofilms suggests potential health risks for consumers., (Copyright © 2018 Elsevier B.V. All rights reserved.)

Understanding a pathogen's response to food environments is imperative to develop effective control strategies as well as to elucidate the impact of foods on virulence potential. The purpose of this study was to assess transcriptional response of Listeria monocytogenes after growth in cantaloupe, as well as its impact on survival in synthetic gastric fluid (SGF). The transcriptional profiles of L. monocytogenes grown in cantaloupe or Brain Heart Infusion (BHI) under refrigeration were compared by a custom-designed microarray. A total of 286 and 175 genes were significantly up- and down-regulated, respectively, in L. monocytogenes grown in cantaloupe as compared to BHI (fold change ≥ 2.5 and adj. P < 0.05). The majority of upregulated genes belonged to functions related to amino acid and nucleotide metabolism, flagellar biosynthesis, and iron acquisition, while most downregulated genes belonged to carbohydrate metabolism. Notably, the branched chain amino acid (BCAA: leucine, isoleucine, valine) biosynthesis operon was shown to be highly upregulated as well as the purine and pyrimidine biosynthesis pathways. Transcript levels of several stress- and virulence-related genes were significantly altered, implying an impact of growth in cantaloupe on the virulence potential of L. monocytogenes. Enhanced survival of L. monocytogenes in SGF following growth in cantaloupe further demonstrated the impact of cantaloupe-associated growth on the pathogen's subsequent response to a host relevant stress., (Copyright © 2018. Published by Elsevier Ltd.)

Plants use many natural products to counter pests and diseases in nature. In rice, direct defense mechanisms include broad range of secondary metabolites, such as phenolamides (PA), diterpene phytoalexins, and flavonoid sakuranetin. Recently, accumulation of PAs in rice was shown to be under control of microbial symbionts in honeydew (HD), digestive waste from the rice brown planthopper ( Nilaparvata lugens ; BPH), but whether HD microbiota can also promote diterpene phytoalexins, momilactone A (MoA) and MoB, has not been reported. Here, we demonstrate that crude HD, but not a filtered one, induces MoA and MoB in rice, suggesting the involvement of BPH-HD endosymbionts. Consequently, microbial strains previously isolated from HD could promote MoA and MoB levels in wounded rice leaves, suggesting that rice indeed responds to BPH by cumulative chemical defense that involves both PA and diterpene phytoalexin pathways.

This study evaluated the potential of blue light-emitting diodes (LED) of wavelength 405 and 460 nm in combination with sodium chlorophyllin to inactivate Listeria monocytogenes and Salmonella spp. on cantaloupe rind. A cocktail culture of L. monocytogenes or Salmonella spp. strains was surface inoculated onto cantaloupe rinds to reach a final concentration of 4 log CFU/cm 2 and dip-treated in a 100 μM sodium copper chlorophyllin solution. The cantaloupe samples were then exposed to 405 or 460 nm LEDs at a total dose of 1210 J/cm 2 and 5356 J/cm 2 , respectively, at 4 and 20 °C. Results showed that the antibacterial efficacy against both pathogens on cantaloupe rinds between LED alone and LED with the chlorophyllin were statistically similar with bacterial inactivation ranging from 1.1 to 3 log CFU/cm 2 in most of the cases or the difference was only minimal. A similar inactivation of 3 log CFU/cm 2 was obtained in the case of L. monocytogenes and Salmonella spp. when illuminated by 405 nm LEDs while the inactivation of L. monocytogenes was higher than Salmonella spp. when illuminated by 460 nm LED. The δ value, a modified Weibull model parameter defined as the time (h) taken to reduce the bacterial population by 90%, was computed to compare the inactivation rates of the conditions. It was inferred that illumination with 405 nm LED required a lower δ value than 460 nm LED illumination (P < 0.05) for the inactivation of L. monocytogenes at 20 °C and Salmonella spp. at 4 °C. Thus the findings of this study indicate a promising application of blue LEDs to inactivate these pathogens on the surface of cantaloupe, minimizing the risk of listeriosis and salmonellosis by consumption of cantaloupe., (Copyright © 2018 Elsevier Ltd. All rights reserved.)

Background: Fruits and vegetables have increasingly been related to foodborne outbreaks. Besides surface contamination, a possible internalization of microorganisms into edible parts of plants during growth has already been observed. To examine an actual risk for the consumer, microbial contamination of the rind and pulp of 147 muskmelons from international trade was assessed using cultural and biochemical methods, polymerase chain reaction and matrix-assisted laser desorption/ionization-time of flight mass spectrometry., Results: One hundred percent of the rind samples [3.69-8.92 log colony forming units (CFU) g -1 ] and 89.8% of the pulp samples (maximum load 3.66 log CFU g -1 ) were microbiologically contaminated. Among the 432 pulp isolates, opportunistic and potentially pathogenic bacteria were identified, mainly Staphylococcus spp. (48.9%), Clostridium spp. (42.9%) and Enterobacteriaceae (27.9%). Salmonella spp., Escherichia coli and isolates of the Bacillus cereus group were found on the rind (1.4%, 0.7% and 42.9%, respectively) and in the pulp (0.7%, 1.4% and 4.7%). Clostridium perfringens was isolated from the rind of seven melons., Conclusion: The present study revealed a regularly occurring internal contamination of melons. Possible health risks for consumers because of an occurrence of microorganisms in melon pulp should be considered in future food safety assessments. © 2018 Society of Chemical Industry., (© 2018 Society of Chemical Industry.)

Salmonella enterica is responsible for the highest number of foodborne disease outbreaks pertaining to cantaloupe industry. The objective of this study was to examine the growth and biofilm formation by outbreak strains of S. enterica ser. Poona (S. Poona), S. enterica ser. Stanley (S. Stanley) and S. enterica ser. Montevideo (S. Montevideo) on different food-contact processing surfaces in cantaloupe flesh and peel extracts at 22 °C and 10 °C. The generation time of all S. enterica strains tested was shorter in the high concentration (50 mg/ml) of cantaloupe extract and high temperature. In 50 mg/ml of cantaloupe flesh or peel extract, the populations of S. enterica were increased by 5 log CFU/ml in 24 h at 22 °C and 1 log CFU/ml in 72 h at 10 °C. In 2 mg/ml of cantaloupe flesh or peel extracts, the populations of S. enterica were increased by 3.5 log CFU/ml in 56 h at 22 °C, but there were no changes in 72 h at 10 °C. The biofilm production of S. enterica was greater at 50 mg/ml of cantaloupe extract and 22 °C, but no major differences (P ≥ 0.05) were found among the strains tested. In 50 mg/ml cantaloupe extract, S. enterica produced 5-6 log CFU/cm 2 biofilm in 4-7 d at 22 °C and approximately 3.5-4 log CFU/cm 2 in 7 d at 10 °C. In 2 mg/ml of cantaloupe extract, S. enterica produced 4-4.5 log CFU/cm 2 biofilms in 4-7 d at 22 °C and 3 log CFU/cm 2 in 7 d at 10 °C. Biofilm formation by S. Poona (01A4754) was lowest on buna-n rubber compared to stainless steel, polyethylene and polyurethane surfaces under the majority of conditions tested. Overall, these findings show that S. enterica strains can grow rapidly and form biofilms on different cantaloupe processing surfaces in the presence of low concentrations of cantaloupe flesh or peel extracts., (Copyright © 2018 Elsevier B.V. All rights reserved.)

Field workers often wear gloves harvesting ready-to-eat produce; however, fields are not sterile environments and gloves may become contaminated numerous times during a working shift. This study explored the potential for inactivation of Escherichia coli O157:H7 and Salmonella when contaminated gloves were washed in levulinic acid (LV) and sodium dodecyl sulfate (SDS) solutions. Washing nitrile gloves with increasing concentrations of LV above 1.0% led to a decreased prevalence of glove contamination by Salmonella (P = 0.0000). A higher level of prevalence occurred for solid agar-cultured pathogens than liquid broth-cultured pathogens after nitrile gloves were washed in LV/SDS (P = 0.0000). Pathogens residing on latex gloves were more likely to be completely inactivated by washing in 0.5% LV/0.1% SDS solutions than nitrile or Canners gloves that exhibited inconsistent responses dependent on the pathogen strain. However, drying after washing nitrile gloves in 0.5% LV/0.1% SDS led to additional pathogen inactivation (P = 0.0394). Pathogen transfer from gloves to produce was implied as the pathogen prevalence on cantaloupe rind handled by LV/SDS-washed gloves was not statistically different from the prevalence on gloves (P = 0.7141). Hence, the risk of produce contamination may still exist but would be reduced by washing gloves in LV/SDS., (Copyright © 2018 Elsevier Ltd. All rights reserved.)

At least 24 species of cucurbits from 13 genera are known to be susceptible to gummy stem blight, caused by three species of Stagonosporopsis. Cankers that are formed on crowns and stems play an important role in the disease cycle and the survival of the pathogen. Fourteen cucurbit species of unknown susceptibility representing 12 genera, four taxonomic tribes, and four geographic origins were inoculated with Stagonosporopsis citrulli in Charleston, SC, in spring 2015, spring 2016, and fall 2016 to evaluate their level of susceptibility to gummy stem blight and the ability of the pathogen to reproduce on crown cankers. An additional species, Cucumis melo, was included as a reference due to its known high susceptibility. Data sets of area under the disease progress curve (AUDPC) for foliar severity and crown cankers, final percentage of diseased leaf area, final percentage of plants with cankers, final percentage of plants with fruiting bodies, and rates of increase in canker incidence were analyzed to evaluate susceptibility. Results were similar for datasets of AUDPC and final ratings but there were more differences for AUDPC. In all experiments, Apodanthera sagittifolia, Ecballium elaterium, and Kedrostis leloja were at least as susceptible to foliar blight as the reference C. melo. K. leloja was as susceptible to crown cankers as C. melo in all experiments and A. sagittifolia and E. elaterium were among the species most susceptible to crown cankers in two experiments. Coccinia grandis was highly resistant to gummy stem blight and had a few cankers only in fall 2016. Sicana odorifera and Zehneria pallidinervia also consistently grouped with the most resistant species. Incidence of crown cankers on Cucumis melo and K. leloja increased at the fastest rate of all species in all experiments and had, along with E. elaterium, the highest incidence of crowns with fruiting bodies. In general, the most susceptible species also were most suitable for reproduction of the pathogen and had the fastest disease progression. The tribes Benincaseae and Cucurbiteae had consistently lower levels of foliar blight than Bryonieae and Coniandreae. The tribe Benincaseae had a consistently lower AUDPC for canker incidence than Bryonieae and Coniandreae. The species originating from Europe (E. elaterium) was consistently most susceptible to both symptoms, while African species grouped with the least susceptible species in all experiments. To the best of our knowledge, this is the first report of susceptibility to gummy stem blight of 14 species and the first report of susceptibility of the cucurbit tribes Coniandreae and Gomphogyneae. This expands the host range of Stagonosporopsis citrulli to 37 species representing 21 genera and seven tribes in the family Cucurbitaceae. This study demonstrates the importance of crown cankers as reproductive sites for S. citrulli.

In order to investigate the effect of S-adenosylmethionine decarboxylase (SAMDC) gene in melon resistance to powdery mildew, according to the previous results of EST sequences from cDNA-AFLP library and the melon genome sequence data, the SAMDC gene was isolated from Chinese wild melon clone 'Yuntian930' by RT-PCR and designated as CmSAMDC (GenBank Accession No. KF151861). The mORF (main open reading frame) was 1 095 bp encoding 364 amino acids with a molecular mass of 40 kDa. The predicted protein sequence showed high similarity with Cucumis sativus and Citrofortunella microcarpa. The SDS-PAGE showed that the expression protein was a fusion protein with the molecular weight of 40 kDa, which perfectly matched the mass calculated from the amino acid sequence. Quantitative real-time PCR analysis indicated that the expression level of CmSAMDC reached a maximum at 48 hpi (hours post inoculation) that over 7-fold to 0 hpi, and the expression of CmSAMDC was also detected in tendril, root, leaf and stem. These results indicate that CmSAMDC gene may play protective roles in melon resistance to powdery mildew infection.

The purpose of this study was evaluation of the effectiveness of superheated steam (SHS) on inactivation of foodborne pathogens on cantaloupes and watermelons. Saturated steam (SS) treatment was performed at 100 °C and that of SHS at 150 and 200 °C. Escherichia coli O157:H7, Salmonella Typhimurium and Listeria monocytogenes-inoculated cantaloupes and watermelons were exposed for a maximum of 30 s and 10 s, respectively. Populations of the three pathogens on cantaloupes and watermelons were reduced by more than 5 log after 200 °C steam treatment for 30 s and 10 s, respectively. After SHS treatment of cantaloupes and watermelons for each maximum treatment time, color and maximum load values were not significantly different from those of untreated controls. By using a noncontact 3D surface profiler, we found that surface characteristics, especially surface roughness, is the main reason for differences in microbial inactivation between cantaloupes and watermelons. The results of this study suggest that SHS treatment can be used as an antimicrobial intervention for cantaloupes and watermelons without inducing quality deterioration., (Copyright © 2017 Elsevier Ltd. All rights reserved.)

Infection risk models of downy mildew of cucumber caused by Pseudoperonospora cubensis were evaluated for their performance in predicting the infection risk of squash and cantaloupe plants under field conditions. Experiments were conducted from 2012 to 2014 in Clayton, NC and Charleston, SC, where disease-free potted plants were exposed to weather conditions during a 24- and 48-h period (hereafter 24- and 48-h models, respectively) within a plot with naturally occurring inoculum. Exposed plants were subsequently placed in a growth chamber where they were monitored for disease symptoms, which was indicative of a successful infection. Disease severity was assessed after 7 days as the proportion of leaf area with disease symptoms. Two predictor variables, day temperature and hours of relative humidity >80% during each exposure were used as inputs to generate model predictions that were compared with observed data. The threshold probability on the receiver operating characteristic (ROC) curve that minimized the overall error rate for the 24-h model was 0.85 for both squash and cantaloupe. The 24-h model was consistently more accurate than the 48-h model in predicting the infection risk for the two hosts. The accuracy of the 24-h model as estimated using area under ROC curve ranged from 0.75 to 0.81, with a correct classification rate ranging from 0.69 to 0.74 across the two hosts. Specificity rates for the model ranged from 0.81 to 0.84, while the sensitivity rates ranged from 0.58 to 0.67. Optimal decisions thresholds (P OT ) developed based on estimates of economic damage and costs of management showed that P OT was dependent on the probability of disease occurrence, with the benefit of using the 24-h model for making management decisions being greatest at low levels of probability of disease occurrence. This 24-h model, previously developed using cucumber as the host, resulted in accurate estimates of the daily infection risk of squash and cantaloupe and could potentially be useful when incorporated into a decision support tool to guide fungicide applications to manage downy mildew in these other cucurbit host types.