Stirred Yogurt as a Delivery Matrix for Freeze-Dried Microcapsules of Synbiotic EVOO Nanoemulsion and Nanocomposite.

Nowadays, dairy products are considered a good matrix to deliver many functional substances either vital oils or probiotic cells. Two models of microcapsules were produced from co-encapsulation of extra virgin olive oil (EVOO) nanoemulsion or nanocomposite and synbiotic bacteria (maltodextrin with Lactobacillus acidophilus and Bifidobacterium bifidum) using the freeze-drying technique. These models of microcapsules were added to stirred yogurt, and then its storage effect on microbiology, chemically, and sensory properties were evaluated for 21 days. The average droplet size and zeta potential distribution of EVOO nanoemulsion and nanocomposite were investigated. Also, oxidative stability, microencapsulation efficiency, release profile, and antioxidant activity were studied. The results showed that the average particle size of EVOO nanoemulsion and nanocomposite ranged between 416 and 475 nm, while zeta potential was -39.6 and -33.6 mV, respectively. The induction period of EVOO extracted from nanoemulsion and nanocomposite microcapsules models was 11.30 and 8 h. The microencapsulation efficiency of probiotic and EVOO was determined at 88.84 and 65.61% for the nanoemulsion microcapsules model, while the nanocomposite microcapsules model showed 98.49 and 72%. The two models of microcapsules have boosted the viability of probiotic bacteria inside stirred yogurt than free cells. Also, the presence of microcapsules did not affect the viability of stirred yogurt starter cultures, and high values for the total solid and protein were detected. Therefore, the results recommended that stirred yogurt is a good delivery carrier for highly antioxidant and healthy microcapsules of synbiotic EVOO nanoemulsion and nanocomposite.

Antifungal activity of eco-safe nanoemulsions based on Nigella sativa oil against Penicillium verrucosum infecting maize seeds: Biochemical and physiological traits.

The main goals of the present investigation were to develop O/W nanoemulsion fungicides based on cold-pressed Nigella sativa (black seed) oil to prevent Penicillium verrucosum infection of maize seeds and to test their antifungal activity against this fungus. Additionally, the effect of these nanoemulsions on plant physiological parameters was also investigated. Two nonionic surfactants namely Tween 20 and Tween 80 were used as emulsifying agents in these formulations. The effect of sonication time and surfactant type on the mean droplet size, polydispersity index (PDI), and zeta potential of the nanoemulsions were determined by dynamic light scattering (DLS). Results indicated that both sonication time and emulsifier type had pronounced effects on the stability of O/W nanoemulsions with a small particle size range (168.6-345.3 nm), acceptable PDI (0.181-0.353), and high zeta potential (-27.24 to -48.82 mV). Tween 20 showed superior stability compared to Tween 80 nanoemulsions. The in vitro results showed that complete inhibition of P. verrucosum-growth was obtained by 10_T80 and 10_T20 nanoemulsions at 100% concentration. All nanoemulsions had increment effects on maize seed germination by 101% in the case of 10_T20 and 10_T80 compared to untreated seeds or the chemical fungicide treatment. Nanoemulsions (10_T20 and 10_T80) were able to stimulate root and shoot length as compared to the fungicide treatment. Seed treatment with 10_T80 nanoemulsion showed the highest AI and protease activity by 75 and 70%, respectively, as compared to the infected control. The produced nanoemulsions might provide an effective protectant coating layer for the stored maize seeds.

Chitosan/silica nanocomposite-based formulation alleviated gray mold through stimulation of the antioxidant system in table grapes.

The main purpose of this study was to explore the ability of a novel silica/polysaccharide polymer-based formulation, namely, chitosan/silica nanocomposites (CSNs), to directly affect Botrytis cinerea in vitro and in inoculated berries, and indirectly to induce natural host resistance via enzymatic and nonenzymatic antioxidants against gray mold of table grapes. The results indicated a positive correlation in in vitro tests in terms of radial growth, spore germination and germ tube elongation, where those parameters were completely inhibited by CSN at 1%. SEM and TEM investigations showed that morphological and internal structural damage was observed in B. cinerea-hyphae/spores treated with CSN. Additionally, most of the treated spores were affected, and cellular vacuolization and cytoplasmic disorganization were observed. The results revealed that CSN reduced gray mold incidence and severity on inoculated berries directly and indirectly. In direct activity, CSN (1%) reduced mold incidence and severity by 100% compared to the control. In indirect activity, mold incidence and severity was reduced by 51% and 64%, respectively. CSN significantly increased superoxide dismutase, ascorbate peroxidase, peroxidase, total phenol and flavonoid at 48 h post-treatment by 1.2-, 1.6-, 1.3-, 1.3- and 1.6-fold, respectively, in grape-treated tissues. It could be concluded that CSN, as a promising alternative control method against gray mold of table grapes, can directly affect the pathogen and indirectly enhance the natural host resistance of the antioxidant system.

Salt Solution Treatments Trigger Antioxidant Defense Response against Gray Mold Disease in Table Grapes.

To obtain a clear understanding of the mode of action of potassium bicarbonate (PB), sodium silicate (SSi) and calcium chelate (CCh) solutions (1%) in inducing resistance to gray mold disease in table grapes, enzymatic and nonenzymatic investigations were carried out. In particular, changes in the activity of the enzymes superoxide dismutase (SOD), ascorbate peroxidase (APX) and peroxidase (POD), total phenolic content and total flavonoid content were studied. As indirect action, PB, SSi and CCh reduced the incidence of gray mold by 43%, 50% and 41%, respectively. The highest activity of SOD was detected at 48 h in SSi-treated tissue, PB-treated tissue and CCh-treated tissue, and it was 1.7-, 1.4- and 1.2-fold higher, respectively, compared to the control. The APX activity was significantly higher in SSi-treated tissue than in the control at 24, 48 and 72 h and showed an increase in activity 2-fold for all times. Additionally, PB, SSi and CCh increased the activity of POD by 1.4-, 1.2- and 2.7-fold at 48 h posttreatment, respectively. The results showed that CCh was the most pronounced salt to increase both total phenol and flavonoid contents by 1.3 and 2.1, respectively. Additionally, the three tested salts induced an increase in total phenols and total flavonoids at 48 h posttreatment. The obtained result is one more movement towards an overall understanding of the mechanism by which salt solutions act as antimicrobial agents against gray mold of table grapes.

Genomic Insights Into the Antifungal Activity and Plant Growth-Promoting Ability in Bacillus velezensis CMRP 4490.

The main objective of this study was to evaluate Bacillus velezensis strain CMRP 4490 regarding its ability to inhibit soil-borne plant pathogens and to increase plant growth. The study included evaluation of in vitro antifungal control, sequencing the bacterial genome, mining genes responsible for the synthesis of secondary metabolites, root colonization ability, and greenhouse studies for the assessment of plant growth-promoting ability. The strain was obtained from soil samples in the north of Paraná in Brazil and was classified as a B. velezensis, which is considered a promising biological control agent. In vitro assay showed that B. velezensis CMRP 4490 presented antagonistic activity against Sclerotinia sclerotiorum, Macrophomina phaseolina, Botrytis cinerea, and Rhizoctonia solani with a mycelial growth inhibition of approximately 60%, without any significant difference among them. To well understand this strain and to validate its effect on growth-promoting rhizobacteria, it was decided to explore its genetic content through genome sequencing, in vitro, and greenhouse studies. The genome of CMRP 4490 was estimated at 3,996,396 bp with a GC content of 46.4% and presents 4,042 coding DNA sequences. Biosynthetic gene clusters related to the synthesis of molecules with antifungal activity were found in the genome. Genes linked to the regulation/formation of biofilms, motility, and important properties for rhizospheric colonization were also found in the genome. Application of CMRP 4490 as a coating film on soybean increased from 55.5 to 64% on germination rates when compared to the control; no differences were observed among treatments for the maize germination. The results indicated that B. velezensis CMRP 4490 could be a potential biocontrol agent with plant growth-promoting ability.

Control of Gray Mold on Clamshell-Packaged 'Benitaka' Table Grapes Using Sulphur Dioxide Pads and Perforated Liners.

The use of vented clamshells has become popular in the packaging of grapes for local and international markets. The aim of this study is to evaluate the postharvest preservation of 'Benitaka' table grapes individually packaged in vented clamshells using different types of SO2-generating pads and perforated plastic liners during cold storage. A completely randomized design with four replications in a two-factor arrangement with an additional treatment [(4 × 3) + 1] was used. The trials were carried out under two situations: Artificial or natural infections with Botrytis cinerea, which is the causal agent of gray mold on table grapes. The incidence of gray mold, shattered berries, and stem browning were evaluated at 30 and 45 days of cold storage at 1 ± 1 °C and 3 days of shelf-life at 22 ± 1 °C after the period of cold storage. Mass loss and berry firmness were also examined at the end of the cold storage period. The use of dual-release SO2-generating pads containing 5 or 8 g of a.i. and slow-release pads with 7 g of a.i. was effective in controlling the incidence of gray mold in grapes packaged in vented clamshells and kept under cold storage for up to 45 days. Under these storage conditions, perforated plastic liners with 0.3% ventilation area or micro-perforated liners with 1.0% ventilation area reduced the percentage of mass loss and shattered berries.

Nanomaterials as Alternative Control Means Against Postharvest Diseases in Fruit Crops.

Post-harvest diseases of fruit and vegetables have to be controlled because of the high added value of commodities and the great economic loss related to spoilage. Synthetic fungicides are the first choice worldwide to control post-harvest diseases of fruit and vegetables. However, several problems and constraints related to their use have forced scientists to develop alternatives control means to prevent post-harvest diseases. Physical and biological means, resistance inducers, and GRAS (generally recognized as safe) compounds are the most important alternatives used during the last 20 years. Recently, nanomaterial treatments have demonstrated promising results and they are being investigated to reduce the utilization of synthetic fungicides to control post-harvest rot in fruit and vegetables. The collective information in this review article covers a wide range of nanomaterials used to control post-harvest decays related to each selected fruit crop including grape, citrus, banana, apple, mango, peach, and nectarine. Other examples also used are apricot, guava, avocado, papaya, dragon, pear, longan, loquat, jujubes, and pomegranate fruits.

Ultra-Structural Alterations in Botrytis cinerea-The Causal Agent of Gray Mold-Treated with Salt Solutions.

Potassium bicarbonate (PB), calcium chelate (CCh), and sodium silicate (SSi) have been extensively used as antifungal generally recognized as safe (GRAS) compounds against plant pathogenic fungi. In this research, in in vitro tests, the radial growth, conidial germination, and germ tube elongation of Botrytis cinerea was completely inhibited at 0.3% of PB, SSi, and CCh. In in vivo tests, application of PB, SSi, and CCh completely inhibited the occurrence of gray mold incidence of inoculated 'Italia' grape berries at concentrations of 1.0, 0.8, and 0.8%, respectively. In order to investigate the detailed mechanisms by which salts exhibited antifungal activity, we analyzed their influence on morphological changes by scanning electron microscopy (SEM) and transmission electron microscopy (TEM) and also on reactive species of oxygen (ROS), mitochondrial membrane potential (MMP), and adenosine triphosphate (ATP) content. Defects such as malformation and excessive septation were detected on salt-treated hyphae morphology observed by SEM. The internal structure of conidia treated or not with salt solutions was examined by TEM. In treated conidia, most of the conidia were affected and cellular vacuolization and cytoplasmic disorganization was observed. For ROS accumulation, a higher increase was observed in fluorescent conidia in presence of PB, SSi, and CCh by 75, 68, and 70% as compared to control, respectively. MMP was significantly decreased after salt application indicating a loss of mitochondria function. Also, luminescence showed that B. cinerea-conidia treated with salts contained less ATP than the untreated conidia. The results obtained herein are a step towards a comprehensive understanding of the mode of action by which salts act as antifungal agents against B. cinerea.

Synergistic effect of a novel chitosan/silica nanocomposites-based formulation against gray mold of table grapes and its possible mode of action.

Ecofriendly nanostructured materials have been proposed as promising alternative control means to prevent plant diseases. Chitosan nanoparticles (CN), silica nanoparticles (SN) and chitosan-silica nanocomposites (CSN) were synthesized and their morphology and structure was characterized by transmission electron microscope (TEM), scanning electron microscopy (SEM), infrared spectra (FT-IR) and Raman spectroscopy. Their antifungal efficiency against Botrytis cinerea, the causal fungus of gray mold disease of table grapes, was tested in vitro and in vivo (under artificial and natural infections). In vitro tests showed that CN, SN and CSN reduced fungal growth by 72, 76 and 100%, respectively at 1% as compared to control. Under natural infection, at the end of cold storage, CSN was the most effective treatment, and reduced the development of gray mold by 59 and 83%, for 'Italia' and 'Benitaka' grapes, respectively as compared to the water control. Results indicate that a synergistic effect of CSN against gray mold was observed. The impact of tested nanocomposites on soluble solids - TSS, titratable acidity - TA, TSS/TA, berry color, mass loss, stem browning and shattered berries was investigated. No negative effect of tested nanomaterials in term of grape quality was observed. For 'Italia' table grape, CN and CSN can preserve bunches from mass loss as compared to control. Also, the effect of CSN on reactive species of oxygen (ROS), ATP content and mitochondrial membrane potential (MMP) of B. cinerea spores was determined to verify its mode of action. The obtained results suggested CSN, as alternative control means, to reduce/substitute the use of fungicides to control gray mold of table grapes while maintaining grape quality.