Post-harvest biocontrol of Fusarium infection in tomato fruits using bio-mediated selenium nanoparticles.

The protection of post-harvest infection by Fusarium spp. is a major worldwide demand, especially using effective natural alternatives to chemical fungicides. In this respect, selenium nanoparticles (SeNPs) were biosynthesized using Fenugreek seeds aqueous extract. Bio-mediated SeNPs were characterized using XRD, FTIR, UV-Vis, TEM, and EDX. XRD confirmed the crystalline nature with six characteristic peaks corresponding to Se-nanocrystals. TEM showed spherical-shaped SeNPs with 34.02-63.61 nm diameter. FTIR verified the presence of different bio-functional groups, such as, N-H, O-H, C-N, and C-NH2 acting as stabilizing/reducing agents during the biosynthesis. Bio-mediated SeNPs exhibited excellent biocidal activity against F. oxysporum and F. moniliforme, with MIC of 0.25 and 1.7 mg/mL, respectively. Direct treatment of F. oxysporum with SeNPs led to significant deformation and lysis of the fungal hyphae within 18 h. The treatment of infected fruits with MIC of SeNPs reduced the infection signs by 100% and preserved the fresh-like appearance of treated fruits for 25 and 35 days when stored at 25 °C and 5 °C, respectively. Therefore, SeNPs is considered efficacious fungicidal against Fusarium spp. in-vitro and in-vivo. The treatment of tomato fruits with MIC of SeNPs positively affected its chemical properties, as well as decreased weight loss %, confirming the barrier effect of SeNPs, thus increasing fruits' shelf-life. Bio-mediated SeNPs appeared safe towards normal HSF and OEC cell lines with IC50> 300 µg/mL. Overall results recommend the usage of bio-mediated SeNPs as safe powerful bioagent against Fusarium infection, maintaining tomato quality, and providing protection from post-harvest invasion and/or destroying existing infections.

Microbially-Induced-Calcite-Precipitation (MICP): A biotechnological approach to enhance the durability of concrete using Bacillus pasteurii and Bacillus sphaericus.

Developing bio-based self-healing concrete aims to minimize durability problems related to cracking. In this study, MICP was used as a smart and eco-friendly approach to produce bio-based durable materials. Bacillus pasteurii (BP) and Bacillus sphaericus (BS) were added into mortar mixtures with 0.25% and 0.5% cement weight. All treated samples exhibited a significant decline in water uptake, capillary permeability, and volume of permeable voids, as compared to control with no bacteria. All treated samples showed significant increase in compressive strength by 28-50%, after 28 days of curing. At the age of 120 days, the flexural strength of all treated samples was significantly increased by 19.29-65.94%. SEM imaging and EDAX confirmed that treated samples were denser with less voids due to MICP. DTA verified that the calcite amount and the crystallinity degree were improved in treated samples. Load deflection of bacterial Reinforced-Laminates had less deformation than control. Reloaded bacterial Reinforced-Laminates exhibited excellent restoration of physico-mechanical properties and performance, after 28, 90, and 120 days, confirming the healing process. Microbial self-healing is an innovative approach for continuous repair of micro-cracks in concrete, improving its durability, thus can reduce the maintenance costs.

Characterization, cytotoxicity, and genotoxicity properties of novel biomediated nanosized-silver by Egyptian Streptomyces roseolus for safe antimicrobial applications.

Multi-drug resistant bacteria are not effectively managed with current treatments, making it a serious global problem. Therefore, there is an essential need for finding new antimicrobial agents. In this regard, silver nanoparticles (Ag-NPs) have been projected as a new generation of antimicrobial agents. Ag-NPs were biomediated by Egyptian Streptomyces roseolus for the first time, which was molecularly identified using 16S rRNA sequencing under accession no. MT071505. Biosynthesized Ag-NPs were characterized using UV-Vis spectroscopy, XRD, TEM, FTIR, and DLS. FTIR analysis confirmed the presence of different bioactive functional groups, such as N-H, C-H, C-O-C, C-NH2, and C=O acting as bioreducing/stabilizing agents for Ag-NPs. Ag-NPs exhibited antimicrobial activity against some multi-drug resistant Gram-positive and Gram-negative pathogens. MBC of Ag-NPs against Listeria monocytogenes and Klebsiella pneumonia were 0.195 and 0.048 mg/mL, respectively, with a tolerance level of 2 confirming its biocidal effect. SEM imaging of Ag-NPs-treated L. monocytogenes and K. pneumonia showed shrunk destroyed cells after 6 h of treatment. Biosynthesized Ag-NPs exhibited IC50 of < 0.3 and 8.21 mg/mL, on normal Human Skin Fibroblast, and Blood Lymphocytes, respectively. IC50 values were significantly higher than its MBC values, with no harmful cytotoxic effect, thus can be safely applied at its biocidal concentration. For biosafety purposes, the genotoxicity of biosynthesized Ag-NPs was assessed using Comet assay for the first time on Blood Lymphocytes, with zero-tail and 100% head intensity indicating non-genotoxic effect. An ecofriendly biomediated synthesis of Ag-NPs was described with easy scale-up, non-toxic by-products, so, it can be recommended as a powerful-safe antimicrobial agent.

Biosynthesis of polyunsaturated fatty acids by two newly cold-adapted Egyptian marine yeast.

The widespread awareness of polyunsaturated fatty acids (PUFAs) benefits for human health has increased the need for their commercial production. Two oleaginous yeast were isolated from the Mediterranean Sea fish and Red Sea fish Epinephelus aeneus and E. areolatus, respectively. These marine candidates were identified by MALDI-TOF/MS biotyper® as Lodderomyces elongisporus and Rhodotorula mucilaginosa. The effect of incubation temperature (7, 15, and 26 °C) and glucose concentration (3% and 8%) on their lipids content were investigated using Sulfo-Phospho-Vanillin (SPV) assay. Their intercellular lipids were visualized by fluorescence microscope using Nile-Red dye. L. elongisporus and R. mucilaginosa produced 20.04% and 26.79% of Linoleic acid, respectively, on normal Basal-Defatted Medium (BDM). Linoleic acid (21.4-22.7%) and α-Linolenic acid (7.5-10.8%) were produced by R. mucilaginosa and L. elongisporus, on normal BDM at 15 °C. High-Glucose BDM induced a positive effect on the total lipids production that reached its maximum of 48% and 54% by R. mucilaginosa and L. elongisporus, respectively, grown at 15 °C. Remarkably, 12.12% of long-chain 15-Docosenoic acid (C22:1) and 21.49% of Tricosanoic acid (C23:0) were detected in the FAs profile of L. elongisporus, when grown on normal BDM at 26 °C. The present study is the first one reporting the FAs profile of the Egyptian Marine L. elongisporus, and its capability to accumulate high amounts of lipids under appropriate fermentation conditions; thus, it could be considered for scaling up production. SUPPLEMENTARY INFORMATION: The online version contains supplementary material available at 10.1007/s13205-021-03010-4.

Bioprocess optimization of glutathione production by Saccharomyces boulardii: biochemical characterization of glutathione peroxidase.

The well-known probiotic GRAS Saccharomyces boulardii (CNCM I-745) was used for the first time to produce glutathione (GSH). The culture conditions affecting GSH biosynthesis were screened using a Plackett-Burman design (PBD). Analyzing the regression coefficients for 12 tested variables, yeast extract, glucose, peptone, cysteine, temperature and agitation rate had a positive significant effect on GSH production with a maximum yeild 192 mg/L. The impact of kinetics of adding cysteine was investigated in 19 experiments during the growth time course (0-36 h), and the maximum yield of glutathione (235 mg/L) was obtained by addition of cysteine after 8 h post-inoculation. The most significant variables were further explored at five levels using central composite rotatable design (CCRD), giving a maximum production of GSH (552 mg/L). Using baffled flasks, the yield of GSH was increased to 730 mg/L, i.e., 1.32-fold increment. The two rate-limiting genes of GSH biosynthesis "γ-glutamyl cysteine synthetase (GSH1) and GSH-synthetase (GSH2)" were amplified and sequenced to validate the GSH biosynthetic potency of S. boulardii. The sequences of genes showed 99% similarity with GSH1 and GSH2 genes of S. cerevisiae. Glutathione peroxidase was purified and characterized from S. boulardii with molecular mass and subunit structure of 80 kDa and 35 kDa as revealed from native and SDS-PAGE, ensuring its homodimeric identity. The activity of GPx was reduced by 2.5-fold upon demetallization confirming its metalloproteinic identity. The GPx was strongly inhibited by hydroxylamine and DTNB, ensuring the implication of surface lysine and cysteine residues on the enzyme active site domains.

Production of bioconcrete with improved durability properties using Alkaliphilic Egyptian bacteria.

Microbial-based self-healing of concrete represents innovative technology for improving micro-crack sealing. Microbial bioactivity can induce calcite-precipitation in concrete, which seals micro-cracks. In this respect, two Egyptian bacterial isolates were selected and identified, as Bacillus subtilis (Bs) and Bacillus megaterium (Bm) using MALDI-TOF/MS-Biotyper®. Peak patterns of the bacterial ribosomal proteins showed a high match between samples and standards, which verified species consistency. Bs and Bm were added to the mortar mixture in two concentrations (0.5%, 1%) of cement weight, then the mechanical and physical properties were tested throughout a 180-day time course. The compressive strength of Bm0.5 bacterial mortar samples was increased by 21.4% after 28 days, as compared to control. The rate of water absorption of Bm samples was decreased by 12.4% after 180 days. Bacterial mortar samples showed significant restoration of compressive strength than the original samples by 44%, 21%, and 52.6% for Bs1, Bm0.5, and Bs0.5, respectively. SEM and EDAX analyses confirmed that bacterial samples were denser with fewer voids than the control, as a result of microbial nanosized calcite-precipitation. DTA verified that the amount of CaCO3 and its degree of crystallinity were increased in the bacterial mortar samples. Load-deflection of reinforced-laminates for bacterial mortar samples showed ductile behavior and less deformation as compared to control. In this work, novel concrete with improved mechanical and physical properties has been developed using selected Egyptian microorganisms, it can promote self-healing of micro-cracks with improved durability of the concrete. The application of self-healing bioconcrete can reduce the inspection and maintenance costs. SUPPLEMENTARY INFORMATION: The online version contains supplementary material available at 10.1007/s13205-021-02781-0.

Trichosporon jirovecii-mediated synthesis of cadmium sulfide nanoparticles.

Cadmium sulphide is one of the most promising materials for solar cells and of great interest due to its useful applications in photonics and electronics, thus the development of bio-mediated synthesis of cadmium sulphide nanoparticles (CdS NPs) is one of the essential areas in nanoparticles. The present study demonstrates for the first time the eco-friendly biosynthesis of CdS NPs using the yeast Trichosporon jirovecii. The biosynthesis of CdS NPs were confirmed by UV-Vis spectrum and characterized by X-ray diffraction assay and electron microscopy. Scanning and transmission electron microscope analyses shows the formation of spherical CdS NPs with a size range of about 6-15 nm with a mean Cd:S molar ratio of 1.0:0.98. T. jirovecii produced hydrogen sulfide on cysteine containing medium confirmed by positive cysteine-desulfhydrase activity and the colony color turned yellow on 0.1 mM cadmium containing medium. T. jirovecii tolerance to cadmium was increased by the UV treatment and three 0.6 mM cadmium tolerant mutants were generated upon the UV radiation treatment. The overall results indicated that T. jirovecii could tolerate cadmium toxicity by its conversion into CdS NPs on cysteine containing medium using cysteine-desulfhydrase as a defense response mechanism.

Foodborne Pathogens Prevention and Sensory Attributes Enhancement in Processed Cheese via Flavoring with Plant Extracts.

Cheese contaminations with foodborne bacterial pathogens, and their health outbreaks, are serious worldwide problems that could happen from diverse sources during cheese production or storage. Plants, and their derivatives, were always regarded as the potential natural and safe antimicrobial alternatives for food preservation and improvement. The extracts from many plants, which are commonly used as spices and flavoring agents, were evaluated as antibacterial agents against serious foodborne pathogens, for example Listeria monocytogenes, Salmonella Typhimurium, Staphylococcus aureus, and Escherichia coli O157:H7, using qualitative and quantitative assaying methods. Dairy-based media were also used for evaluating the practical application of plant extracts as antimicrobial agents. Most of the examined plant extracts exhibited remarkable antibacterial activity; the extracts of cinnamon, cloves, garden cress, and lemon grass were the most powerful, either in synthetic or in dairy-based media. Flavoring processed cheese with plant extracts resulted in the enhancement of cheese sensory attributes, for example odor, taste, color, and overall quality, especially in flavored samples with cinnamon, lemon grass, and oregano. It can be concluded that plant extracts are strongly recommended, as powerful and safe antibacterial and flavoring agents, for the preservation and sensory enhancement of processed cheese.

Lipase-catalyzed synthesis of structured phenolic lipids in solvent-free system using flaxseed oil and selected phenolic acids as substrates.

Structured phenolic lipids (PLs) were obtained by lipase-catalyzed transesterification of flaxseed oil, in a solvent-free system (SFS), with selected phenolic acids, including hydroxylated and/or methoxylated derivatives of cinnamic, phenyl acetic and benzoic acids. A bioconversion yield of 65% was obtained for the transesterification of flaxseed oil with 3,4-dihydroxyphenyl acetic acid (DHPA). However, the effect of the chemical structure of phenolic acids on the transesterification of flaxseed oil in SFS was of less magnitude as compared to that in organic solvent system (OSS). Using DHPA, the APCI-MS analysis confirmed the synthesis of monolinolenyl, dilinolenyl, linoleyl linolenyl and oleyl linolenyl dihydroxyphenyl acetates as phenolic lipids. A significant increase in the enzymatic activity from 200 to 270 nmol of PLs/g solid enzyme/min was obtained upon the addition of the non-ionic surfactant Span 65. However, upon the addition of the anionic surfactant, sodium bis-2-ethylhexyl sulfosuccinate (AOT), and the cationic one, hexadecyltrimethylammonium bromide (CTAB), the enzymatic activity was decreased slightly from 200 to 192 and 190 nmol of PLs/g solid enzyme/min, respectively. The results also showed that the increase in DHPA concentration from 20 to 60 mM resulted in a significant increase in the volumetric productivity (P(V)) from 1.61 to 4.74 mg PLs per mL reaction mixture per day.

Biological control of bacterial spot of tomato caused by Xanthomonas campestris pv. vesicatoria by Rahnella aquatilis.

Xanthomonas campestris pv. vesicatoria strain 2 was isolated from infected tomato seedlings grown in open field in Egypt. This strain produced irregular yellow-necrotic areas on tomato leaves and spotting of the stem. In an attempt to control this disease biologically, four experiments were conducted and tomato seedlings were pretreated, before the pathogen, with either of two antagonistic strains of Rahnella aquatilis through leaves, roots, soil or seeds. In all experiments, seedlings pretreated with R. aquatilis showed reduced susceptibility toward X. c. pv. vesicatoria. They also contained reduced protein concentration and showed reduced number of protein bands in SDS-PAGE analysis as well as increased fresh and dry weight relative to control seedlings inoculated with the pathogen only. This indicates that R. aquatilis reduced the deleterious effect and the stress exerted by X. c. pv. vesicatoria on tomato seedlings. Foliar application of R. aquatilis was the most effective method in disease reduction which could be attributed to the direct effect of the antagonistic bacteria on the pathogen. The highest amounts of fresh and dry weight were obtained from seed treatment, which might suggest that bacterial seed inoculation provides earlier protection than could be achieved with foliar, soil or root treatment.