Effects of organic and chemical nitrogen fertilization and postharvest treatments on the visual and nutritional quality of fresh-cut celery (Apium graveolens L.) during storage.

The shelf life of horticultural commodities depends on pre- and postharvest factors, such as soil fertilization and postharvest handling. The current study aimed to evaluate fresh-cut celery's postharvest quality as affected by the rate and type (organic and chemical) of nitrogen (N) fertilizer and postharvest treatments. Celery ('Tall Utah') crop was grown in a field in Karaj, Iran. The experimental design was a randomized complete block with three replications and seven preharvest (fertilizer), and five postharvest treatments. Organic fertilizers were vermicompost (VER) and bio-organic fertilizer [farmyard and livestock manure plus Trichoderma harzianum (COM)]. Chemical fertilizers were urea (46% N) at high rate [322 kg·ha1 N (UREA_HIGH)], optimal rate [196 kg·ha-1 N (UREA_OPT)], and low rate [138 kg·ha-1 N (UREA_LOW)]; ammonium nitrate [35% N (AN)] at 196 kg·ha-1 N; and treatment without fertilization was used as a control. Postharvest treatments included plastic packaging (PP), hydrocooling (HC), blanching (B), and edible coating of psyllium seed mucilage (EC). After postharvest treatments, celery petioles were stored (0-2°C, 85%-90% RH) for 4 weeks and evaluated weekly for quality attributes. Organic fertilizers and UREA_LOW were the most effective treatments in reducing the changes in color, weight loss, titratable acidity (TA), pH, and total soluble solids (TSS) of fresh-cut celery. Organic fertilizers enhanced the vitamin C content, total phenols, and antioxidant activity in celeries. As postharvest treatments, hydrocooling, plastic packaging, and blanching maintained chroma and hue values. Blanching had the greatest effect on the L* value. Hydrocooling increased celery's TA, TSS, and vitamin C content and reduced weight loss and pH during storage. Thus, celery quality was improved when grown under low or adequate N fertilization. Hydrocooling was an effective postharvest treatment for preserving fresh-cut celery quality during storage.

Protein signatures to identify the different genera within the Xanthomonadaceae family.

The Xanthomonadaceae family comprises the genera Xanthomonas and Xylella, which include plant pathogenic species that affect economically important crops. The family also includes the plant growth-promoting bacteria Pseudomonas geniculata and Stenotrophomonas rhizophila, and some other species with biotechnological, medical, and environmental relevance. Previous work identified molecular signatures that helped to understand the evolutionary placement of this family within gamma-proteobacteria. In the present study, we investigated whether insertions identified in highly conserved proteins may also be used as molecular markers for taxonomic classification and identification of members within the Xanthomonadaceae family. Four housekeeping proteins (DNA repair and replication-related and protein translation enzymes) were selected. The insertions allowed discriminating phytopathogenic and plant growth-promoting groups within this family, and also amino acid sequences of these insertions allowed distinguishing different genera and, eventually, species as well as pathovars. Moreover, insertions in the proteins MutS and DNA polymerase III (subunit alpha) are conserved in Xylella fastidiosa, but signatures in DNA ligase NAD-dependent and Valyl tRNA synthetase distinguish particular subspecies within the genus. The genus Stenotrophomonas and Pseudomonas geniculata could be distinguishable based on the insertions in MutS, DNA polymerase III (subunit alpha), and Valyl tRNA synthetase, although insertion in DNA ligase NAD-dependent discriminates these bacteria at the species level. All these insertions differentiate species and pathovars within Xanthomonas. Thus, the insertions presented support evolutionary demarcation within Xanthomonadaceae and provide tools for the fast identification in the field of these bacteria with agricultural, environmental, and economic relevance.

Survival of Salmonella enterica and Escherichia coli O157:H7 Sprayed onto the Foliage of Field-Grown Cabbage Plants.

To reduce the number of cabbage pathogen outbreaks, it is essential to understand the fate of enteric pathogens that contaminate plants in the field. To assist in that effort, two independent trials were conducted with a red cultivar (cv. Red Dynasty) and a green cultivar (cv. Bravo F1) of field-grown cabbage ( Brassica oleracea var. capitata). In the first trial, plants with small heads were sprayed with an inoculum containing both attenuated Salmonella enterica Typhimurium and Escherichia coli O157:H7 (5.0 log CFU/mL). Initial pathogen levels (ca. 3.9 log CFU per head), determined through plate count enumeration (limit of detection was 1.3 log CFU/g), dropped precipitously such that 2 days later, they could not be detected by enrichment culture in 22 to 35% of the heads. However, subsequent declines were at a slower rate; no differences were observed between red and green cabbage heads ( P > 0.05), and heads were still positive for the pathogens 22 days after being sprayed with the inoculum. As a result, the logistic model revealed that for every 2 days contaminated cabbage heads remained in the field, the probability of finding a positive sample decreased by a factor of 1.1 (95% confidence interval from 1.0 to 1.2, P = 0.0022) and 1.2 (95% confidence interval from 1.0 to 1.4, P ≤ 0.0001) for Salmonella and E. coli O157:H7, respectively. In the second trial occurring 2 weeks later, plants with medium red or green cabbage heads were sprayed with an inoculum at a dose of 3.5 log CFU/mL. A similar decay in prevalence over time occurred for green cabbage as in trial 1; however, pathogen decline in red cabbage was less in trial 2 than in trial 1. The extended persistence of pathogens in cabbage heads exhibited in both trials infers that harvest of contaminated cabbage destined for raw consumption is risky. Additional field studies are necessary to determine whether similar pathogen fates occur in other regions or climates and to clarify the effect of the maturity of red cabbage on pathogen inactivation.

Disposition of Salmonella and Escherichia coli O157:H7 following Spraying of Contaminated Water on Cucumber Fruit and Flowers in the Field.

Cucumbers are frequently consumed raw and have been implicated in several recent foodborne outbreaks. Because this item may become contaminated at the farm, it is vital to explore the fate of attenuated Salmonella Typhimurium or Escherichia coli O157:H7 sprayed onto foliage, flowers, and fruit in fields and determine whether pre- or postcontamination spray interventions could minimize contamination. After spraying cucumber plants with contaminated irrigation water (3.8 log CFU/mL of Salmonella Typhimurium and E. coli O157:H7), 60 to 78% of cucumber fruit were not contaminated because the plant's canopy likely prevented many of the underlying fruit from being exposed to the water. Subsequent exposure of contaminated cucumber plants to a simulated shower event did not appear to dislodge pathogens from contaminated foliage onto the fruit, nor did it appear to consistently wash either pathogen from the fruit. Spraying flowers and attached ovaries directly with a pathogen inoculum (4.6 log CFU/mL) initially led to 100% and 65 to 90% contamination, respectively. Within 3 days, 30 to 40% of the flowers were still contaminated; however, contamination of ovaries was minimal (≤10%), suggesting it was unlikely that internalization occurred through the flower to the ovary with these pathogen strains. In another study, both pathogens were found on a withered flower but not on the fruit to which the flower was attached, suggesting that this contaminated flower could serve as a source of cross-contamination in a storage bin if harvested with the fruit. Because pre- and postcontamination acetic acid-based spray treatments failed to reduce pathogen prevalence, the probability that fruit initially contaminated at 1.3 to 2.8 log CFU of Salmonella Typhimurium or E. coli O157:H7 per cucumber would be positive by enrichment culture decreased by a factor of 1.6 and 1.9 for Salmonella Typhimurium and E. coli O157:H7, respectively, for every day the fruit was held in the field ( P ≤ 0.0001). Hence, to reduce the prevalence of Salmonella Typhimurium on cucumbers below 5%, more than 1 week would be required.

Physical and chemical properties of pomegranate fruit accessions from Croatia.

The objective was to evaluate physical and chemical properties of eight pomegranate accessions (seven cultivars and one wild genotype) collected from the Mediterranean region of Croatia. Accessions showed high variability in fruit weight and size, calyx and peel properties, number of arils per fruit, total aril weight, and aril and juice yield. Variables that define sweet taste, such as low total acidity (TA; 0.37-0.59%), high total soluble solids content (TSS; 12.5-15.0%) and their ratio (TSS/TA) were evaluated, and results generally aligned with sweetness classifications of the fruit. Pomegranate fruit had a high variability in total phenolic content (1985.6-2948.7 mg/L). HPLC-MALDI-TOF/MS analysis showed that accessions with dark red arils had the highest total anthocyanin content, with cyanidin 3-glucoside as the most abundant compound. Principal component analysis revealed great differences in fruit physical characteristics and chemical composition among pomegranate accessions.

Absence of internalization of Escherichia coli O157:H7 into germinating tissue of field-grown leafy greens.

Both growth chamber and field studies were conducted to investigate the potential for Escherichia coli O157:H7 to be internalized into leafy green tissue when seeds were germinated in contaminated soil. Internalized E. coli O157:H7 was detected by enrichment in both spinach (Spinacia oleracea L.) and lettuce (Lactuca sativa L.) seedlings when seeds were germinated within the growth chamber in autoclaved and nonautoclaved soil, respectively, contaminated with E. coli O157:H7 at 2.0 and 3.8 log CFU/g, respectively. Internalized E. coli O157:H7 populations could be detected by enumeration within leafy green tissues either by increasing the pathogen levels in the soil or by autoclaving the soil. Attempts to maximize the exposure of seed to E. coli O157:H7 by increasing the mobility of the microbe either through soil with a higher moisture content or through directly soaking the seeds in an E. coli O157:H7 inoculum did not increase the degree of internalization. Based on responses obtained in growth chamber studies, internalization of E. coli O157:H7 surrogates (natural isolates of Shiga toxin-negative E. coli O157:H7 or recombinant [stx- and eae-negative] outbreak strains of E. coli O157:H7) occurred to a slightly lesser degree than did internalization of the virulent outbreak strains of E. coli O157:H7. The apparent lack of internalized E. coli O157:H7 when spinach and lettuce were germinated from seed in contaminated soil (ca. 3 to 5 log CFU/g) in the field and the limited occurrence of surface contamination on the seedlings suggest that competition from indigenous soil bacteria and environmental stresses were greater in the field than in the growth chamber. On the rare occasion that soil contamination with E. coli O157:H7 exceeded 5 log CFU/g in a commercial field, this pathogen probably would not be internalized into germinating leafy greens and/or would not still be present at the time of harvest.

Internalization of Escherichia coli O157:H7 following spraying of cut shoots when leafy greens are regrown for a second crop.

Both spinach and lettuce were grown to harvest, cut, and then regrown after spraying the cut shoots with irrigation water contaminated with Escherichia coli O157:H7. Plant tissue was collected on the day of spraying and again 2 and 14 days later for analysis of total and internalized E. coli O157:H7 populations. Internalization of E. coli O157:H7 occurred on the day of spraying, and larger populations were internalized as the level in the spray increased. Tissue repair was slow and insufficient to prevent infiltration of E. coli O157:H7; internalized E. coli O157:H7 in shoots cut 5 days prior to exposure to E. coli O157:H7-contaminated water were not significantly different from levels in shoots cut on the same day of spraying with contaminated water (P > 0.05). Two days after spraying plants with a high level of E. coli O157:H7 (7.3 log CFU/ml), levels of internalized E. coli O157:H7 decreased by ca. 2.6 and 1.3 log CFU/g in Tyee and Bordeaux spinach, respectively, whereas populations of internalized E. coli O157:H7 decreased very little (ca. 0.4 log CFU/g) in lettuce plants that had been sprayed either on the same day as cutting or 1 day after cutting. When cut plants were sprayed with irrigation water at a lower contamination level (4.5 log CFU/ml), internalized E. coli O157:H7 was not detected in either spinach or lettuce plants 2 days later and therefore would not likely be of concern when the crop was harvested.

Surface and internalized Escherichia coli O157:H7 on field-grown spinach and lettuce treated with spray-contaminated irrigation water.

Numerous field studies have revealed that irrigation water can contaminate the surface of plants; however, the occurrence of pathogen internalization is unclear. This study was conducted to determine the sites of Escherichia coli O157:H7 contamination and its survival when the bacteria were applied through spray irrigation water to either field-grown spinach or lettuce. To differentiate internalized and surface populations, leaves were treated with a surface disinfectant wash before the tissue was ground for analysis of E. coli O157:H7 by direct plate count or enrichment culture. Irrigation water containing E. coli O157:H7 at 10(2), 10(4), or 10(6) CFU/ml was applied to spinach 48 and 69 days after transplantation of seedlings into fields. E. coli O157:H7 was initially detected after application on the surface of plants dosed at 10(4) CFU/ml (4 of 20 samples) and both on the surface (17 of 20 samples) and internally (5 of 20 samples) of plants dosed at 10(6) CFU/ml. Seven days postspraying, all spinach leaves tested negative for surface or internal contamination. In a subsequent study, irrigation water containing E. coli O157:H7 at 10(8) CFU/ml was sprayed onto either the abaxial (lower) or adaxial (upper) side of leaves of field-grown lettuce under sunny or shaded conditions. E. coli O157:H7 was detectable on the leaf surface 27 days postspraying, but survival was higher on leaves sprayed on the abaxial side than on leaves sprayed on the adaxial side. Internalization of E. coli O157:H7 into lettuce leaves also occurred with greater persistence in leaves sprayed on the abaxial side (up to 14 days) than in leaves sprayed on the adaxial side (2 days).

Infrequent internalization of Escherichia coli O157:H7 into field-grown leafy greens.

Several sources of contamination of fresh produce by Escherichia coli O157:H7 (O157) have been identified and include contaminated irrigation water and improperly composted animal waste; however, field studies evaluating the potential for internalization of O157 into leafy greens from these sources have not been conducted. Irrigation water inoculated with green fluorescent plasmid-labeled Shiga toxin-negative strains (50 ml of 10(2), 10(4), or 10(6) CFU of O157 per ml) was applied to soil at the base of spinach plants of different maturities in one field trial. In a second trial, contaminated compost (1.8 kg of 10(3) or 10(5) CFU of O157 per g) was applied to field plots (0.25 by 3.0 m) prior to transplantation of spinach, lettuce, or parsley plants. E. coli O157:H7 persisted in the soil up to harvest (day 76 posttransplantation) following application of contaminated irrigation water; however, internalized O157 was not detected in any spinach leaves or in roots exposed to O157 during the early or late growing season. Internalized O157 was detected in root samples collected 7 days after plants were contaminated in mid-season, with 5 of 30 samples testing positive for O157 by enrichment; however, O157 was not detected by enrichment in surface-disinfected roots on days 14 or 22. Roots and leaves from transplanted spinach, lettuce, and parsley did not internalize O157 for up to 50 days in the second trial. These results indicate that internalization of O157 via plant roots in the field is rare and when it does occur, O157 does not persist 7 days later.

Ripening in papaya fruit is altered by ACC oxidase cosuppression.

Papaya (Carica papaya) is a very important crop in many tropical countries but it is highly susceptible to parasitic diseases, physiological disorders, mechanical damage and fruit overripening. Here we report a study on ACC oxidase cosuppression and its effects on papaya fruit ripening. Papaya ACC oxidase was isolated using PCR and embriogenic cells transformed by biolistic using the CaMV 35S promoter to drive the expression of the PCR fragment in sense orientation. Fifty transgenic lines were recovered and 20 of those were grown under field conditions. Southern analysis showed incorporation of the transgene in different copy numbers in the papaya genome. Fruits were evaluated in terms of texture (firmness), colour development, respiration and ethylene production. A sharp reduction in ethylene and CO2 production was detected, whereas softening and colour development of the peel were also altered. Overall, transgenic fruits showed a delay in ripening rate. A reduction in mRNA level for ACC oxidase in transgenic fruit was clearly detectable by northern blot. More studies are necessary before this technology can be used to extend the shelf life of papaya fruit.