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The rapid and sensitive detection of Salmonella typhimurium in food matrices is crucial for ensuring food safety. This study presents the development of an ultrasensitive electrochemical biosensor using surface-modified bacterial cellulose (BC) integrated with polypyrrole (Ppy) and reduced graphene oxide (RGO), further functionalized with immobilized S. typhimurium-specific phage particles. The BC substrate, with its ultra-fibrous and porous structure, was modified through in situ oxidative polymerization of Ppy and RGO, resulting in a highly conductive and flexible biointerface. The immobilization of phages onto this composite was facilitated by electrostatic interactions between the polycationic Ppy and the negatively charged phage capsid heads, optimizing phage orientation and enhancing bacterial capture efficiency. Morphological and chemical characterization confirmed the successful fabrication and phage immobilization. The biosensor demonstrated a detection limit of 1 CFU/mL for S. typhimurium in phosphate-buffered saline (PBS), with a linear detection range spanning 100 to 107 CFU/mL. In real samples, the sensor achieved detection limits of 5 CFU/mL in milk and 3 CFU/mL in chicken, with a linear detection range spanning 100 to 106 CFU/mL, maintaining high accuracy and reproducibility. The biosensor also effectively discriminated between live and dead bacterial cells, demonstrating its potential in real-world food safety applications. The biosensor performed excellently over a wide pH range (4–10) and remained stable for up to six weeks. Overall, the developed BC/Ppy/RGO–phage biosensor offers a promising tool for the rapid, sensitive, and selective detection of S. typhimurium, with robust performance across different food matrices.

Background: Fungal nanofactories have been utilized to synthesize silver and gold nanoparticles. This study was designed to mycosynthesize and characterize silver and gold nanoparticles (AgNPs and AuNPs) and to study their effect on aflatoxin B1 production by Aspergillus flavus. Results: Silver and gold nanoparticles were synthesized by endophytic Aspergillus versicolor and then analyzed by UV–vis spectroscopy. The results revealed surface plasmon resonance peaks at 432 and 536 nm for Ag and Au nanoparticles, respectively. The obtained transmission electron microscopy results revealed the fashioning of spherical AgNPs and spherical and hexagonal AuNPs with a mean particle magnitude of 5–37 and 37–62 nm, respectively. X-ray diffraction showed the typical face-centered cubic structure of the mycosynthesized Ag and Au nanoparticles. An in vitro investigation showed that AgNPs, AuNPs, and their mixture at different concentrations (10000, 5000, 3000, 1000, 750, 500, 250, and 125 µg/mL) could inhibit or reduce the outgrowth and production of aflatoxin B1 (AFB1) by A. flavus. The concentration that showed no AFB1 production was less than those for the inhibition of fungal growth. AgNPs, AuNPs, and their mixture also exhibited promising antiradical scavenging activity. Conclusions: The use of fungi in the metallic nanoparticle’s fabrication and the utilization of mycosynthesized nanoparticles is promising as a substitute of chemicals to control antiaflatoxigenic fungi.How to cite: Sheikh H, Awad MF. Biogenesis of nanoparticles with inhibitory effects on aflatoxin B1 production by Aspergillus flavus. Electron J Biotechnol 2022;60. https://doi.org/10.1016/j.ejbt.2022.09.003.

Abstract This study aimed to improve lipid and gamma-linolenic acid (GLA) production of an oleaginous fungus, Mucor plumbeus, through coculturing with Bacillus subtilis bacteria, optimising the environmental and nutritional culture conditions, and scaling them for batch fermentation. The maximum levels of biomass, lipid, fatty acid, and GLA in a 5 L bioreactor containing cellobiose and ammonium sulfate as the optimal carbon and nitrogen sources, respectively, achieved during the coculturing processes were 14.5 ± 0.4 g/L, 41.5 ± 1.3, 24 ± 0.8, and 20 ± 0.5%, respectively. This strategy uses cellobiose in place of glucose, decreasing production costs. The nutritional and abiotic factor results suggest that the highest production efficiency is achieved at 6.5 pH, 30 °C temperature, 10% (v/v) inoculum composition, 200 rpm agitation speed, and a 5-day incubation period. Interestingly, the GLA concentration of cocultures (20.0 ± 0.5%) was twofold higher than that of monocultures (8.27 ± 0.11%). More importantly, the GC chromatograms of cocultures indicated the presence of one additional peak corresponding to decanoic acid (5.32 ± 0.20%) that is absent in monocultures, indicating activation of silent gene clusters via cocultivation with bacteria. This study is the first to show that coculturing of Mucor plumbeus with Bacillus subtilis is a promising strategy with industrialisation potential for the production of GLA-rich microbial lipids and prospective biosynthesis of new products.

Background: Endophytic fungi are associated within plant tissues without any harmful effects. They produce a diversity of metabolites and are well known as rich source of bioactive substances. This study aimed to identify fungal isolates from different medicinal plants, evaluate their antioxidant potency and detect components responsible for antioxidant activity. Results: In the present investigation, 4 endophytic fungal isolates were obtained from different medicinal plants Rumex nervosus, Pulicaria crispa and Withania somnifera. Those isolates were characterized as Penicilluim commune, Penicilluim glaucoroseum, Aspergillus flavipes and Fusarium chlamydosporum according to morphological and phylogenetic bases. These strains were propagated on humified rice and their crude ethyl acetate extracts were assayed for their total phenols, flavonoids and antioxidant potency against DPPH free radicals. The results showed that the elevated constituent of phenolics was found in P. commune (148.24 mg/g), followed by F. chlamydosporum (124.25 mg/g), while the highest total flavonoids were determined in A. flavipes, followed by F. chlamydosporum, P. glaucoroseum, P. commune, respectively. Moreover, the maximum hydroxyl radical-scavenging assay was IC50 = 2.13 mg/mL exhibited by P. commune, followed by F. chlamydosporum (2.68 mg/mL). Weak antimicrobial and cytotoxic effects were reported by the studied strains against pathogenic isolates and (HepG2) hepatocellular carcinoma cell line. The fungal EtOAc extracts were characterized quantitatively by the incidence of different phenols, flavonoids, and phenolic acids, as detected by HPLC. Conclusions: Endophytic fungi derived from medicinal plants are a promising candidate for the wide industrial production of antioxidant agents with several useful medical and pharmaceutical applications.How to cite: Hassane A. M. A, Taha T. M, Awad M. F, et al. Radical scavenging potency, HPLC profiling and phylogenetic analysis of endophytic fungi isolated from selected medicinal plants of Saudi Arabia. Electron J Biotechnol 2022;58. https://doi.org/10.1016/j.ejbt.2022.05.001

Among the crop production factors, preceding crop and tillage management affect the sustainable use of soil resources and ultimately crop growth and productivity. This study aimed at investigating the impact of preceding winter crops (grass or legume) and different tillage systems on forage yield, quality and nutritive values of three summer grass (Sudan grass, pearl millet and teosinte) and two legume forage crops (cowpea and guar) under arid conditions. The results exhibited that growing forage crops after legumes (as berseem clover) produced the highest fresh and dry forage yields and quality attributes compared with grasses (as wheat) with the exception of crude fiber content, which was decreased. Moreover, tillage practices showed positive impact on forage yields and quality attributes. The maximum forage yields and quality parameters were recorded under conventional tillage (CT) practice compared with reduced tillage (RT) and no-tillage (NT) systems. Among the evaluated crops, the highest yields of fresh forage, dry forage, crude fiber, crude protein and total digestible nutrient were exhibited by grass forage crops (Sudan grass, pearl millet and teosinte), whereas the highest crude protein content and the digestible energy values were produced by legume forage crops (cowpea and guar). The maximum fresh forage, dry forage, crude fiber, crude protein, total digestible nutrient and digestible crude protein yields were produced by pearl millet followed by Sudan grass under CT and RT after berseem clover. The highest net return was recorded by sowing pearl millet after berseem clover and applying CT followed by RT practices, which could be recommended for the commercial production. Moreover, it could be assumed that the combination of growing grass forage crops after legume crops under CT or RT systems could enhance forage crop yield and quality with an improvement in soil properties for sustainable agriculture with low cost and the highest net income.

Caseins determine the physicochemical, physiological, and biological characteristics of milk. Four caseins—alpha-S-1, alpha-S-2, beta, and kappa—were analyzed phylogenetically and in silico and characterized regarding chemical, antimicrobial, and antioxidant features in five dairy animals: Arabian camels, sheep, goats, cattle, and water buffalos. The sequence of full-length amino acids of the four caseins for the five species was retracted from the NCBI GenBank database. Multiple sequence alignment is used to examine further the candidate sequences for phylogenetic analysis using Clustal X and NJ-Plot tools. The results revealed that sheep and goats possess strong similarities (98.06%) because of their common ancestor. The same was observed with cattle and water buffalos (96.25%). The Arabian camel was located in a single subclade due to low similarity in casein residues and compositions with other dairy animals. Protein modeling showed that alpha-S1- and alpha-S2-caseins possess the highest number of phosphoserine residues. The in silico computed chemical properties showed that β-casein recorded highest hydrophobicity index and lowest basic amino acid content, while α-S2-casein showed the opposite. The computed biological parameters revealed that α-S2-casein presented the highest bactericidal stretches. Only Arabian camel β-casein and k-casein showed one bactericidal stretches. The analysis also revealed that β-casein, particularly in Arabian camels, possesses the highest antioxidant activity index. These results support the importance of the bioinformatics resources to determine milk casein micelles' chemical and biological activities.

Oxidative stress is involved in the pathophysiology of multiple health complications, and it has become a major focus in targeted research fields. As known, black seeds are rich sources of bio-active compounds and widely used to promote human health due to their excellent medicinal and pharmaceutical properties. The present study investigated the antioxidant potency of various black seeds from plants and their derived mycoendophytes, and determined the total phenolic and flavonoid contents in different extracts, followed by characterization of major constituents by HPLC analysis. Finally, in silico docking determined their binding affinities to target myeloperoxidase enzymes. Ten dominant mycoendophytes were isolated from different black seed plants. Three isolates were then selected based on high antiradical potency and further identified by ITS ribosomal gene sequencing. Those isolated were Aspergillus niger TU 62, Chaetomium madrasense AUMC14830, and Rhizopus oryzae AUMC14823. Nigella sativa seeds and their corresponding endophyte A. niger had the highest content of phenolics in their n-butanol extracts (28.50 and 24.43 mg/g), flavonoids (15.02 and 11.45 mg/g), and antioxidant activities (90.48 and 81.48%), respectively, followed by Dodonaea viscosa and Portulaca oleracea along with their mycoendophytic R. oryzae and C. madrasense. Significant positive correlations were found between total phenolics, flavonoids, and the antioxidant activities of different tested extracts. The n-butanol extracts of both black seeds and their derived mycoendophytes showed reasonable IC50 values (0.81–1.44 mg/ml) compared to the control with significant correlations among their phytochemical contents. Overall, seventeen standard phenolics and flavonoids were used, and the compounds were detected in different degrees of existence and concentration in the examined extracts through HPLC analysis. Moreover, the investigation of the molecular simulation results of detected compounds against the myeloperoxidase enzyme revealed that, as a targeted antioxidant, rutin possessed a high affinity (−15.3184 kcal/mol) as an inhibitor. Taken together, the black seeds and their derived mycoendophytes are promising bio-prospects for the broad industrial sector of antioxidants with several valuable potential pharmaceutical and nutritional applications.

Klebsiella is a common dangerous pathogen for humans and animals and is widely present in the digestive system. The genus Klebsiella is ubiquitous, as it is endemic to surface water, soil, and sewage. In this study, 70 samples were obtained from soil-dwelling invertebrates from September 2021 to March 2022 from Taif and Shafa in different altitudinal regions of Saudi Arabia. Fifteen of these samples were identified as Klebsiella spp. The Klebsiella isolates were genetically identified as Klebsiella pneumoniae using rDNA sequencing. The antimicrobial susceptibility of the Klebsiella isolates was determined. Amplification of virulence genes was performed using PCR. In this study, 16S rDNA sequencing showed a similarity from 98% to 100% with related K. pneumonia from the NCBI database, and the sequences were deposited in the NCBI GenBank under accession numbers ON077036 to ON077050. The growth inhibition properties of ethanolic and methanolic extracts of the medicinal plant Rhazya stricta’s leaves against K. pneumoniae strains using the minimum inhibitory concentration (MIC) method and disc diffusion were evaluated. In addition, the biofilm inhibitory potential of these extracts was investigated using crystal violet. HPLC analysis identified 19 components divided into 6 flavonoids, 11 phenolic acids, stilbene (resveratrol), and quinol, and revealed variations in the number of components and their quantities between extracts. Both extracts demonstrated interesting antibacterial properties against K. pneumoniae isolates. The 2 extracts also showed strong biofilm inhibitory activities, with percentages of inhibition extending from 81.5% to 98.7% and from 35.1% to 85.8% for the ethanolic and methanolic extracts, respectively. Rhazya stricta leaf extract revealed powerful antibacterial and antibiofilm activities against K. pneumoniae isolates and could be a good candidate for the treatment or prevention of K. pneumonia-related infections.

Background and objectives: Aspergillus keratitis are in the increasing trend and reported as the second most common cause of mycotic keratitis in developing countries. The present study was designed to isolate, identify Aspergillus spp. from the keratits/corneal ulcer patients attending a tertiary care eye hospital, Coimbatore, South India and to assess the minimum inhibitory concentrations (MICs) against ten clinically used first-line antifungal drugs. Methods: A total of seventy-three Aspergillus strains isolated from corneal scrapings were included and assessed for a period of one year. All isolates were identified up to the species level by morphological observations. Antifungal drug susceptibilities were determined against a standard panel of antifungal agents. Conclusions: Five different species of aspergilli, A. flavus (n=53), A. fumigatus (n=14), A. terreus (n=9), A. tamarii (n=6) and A. niger (n=3) were identified based on morphological features. Minimum inhibitory concentration analyses indicated that, voriconazole, natamycin, itraconazole, clotrimazole, econazole followed by ketoconazole shall be the order of choices for the effective treatment for Aspergillus keratitis.

In this study, 18 standard amino acids were tested as a single nitrogen source on biomass, total lipid, total fatty acid (TFA) production, and yield of γ-linolenic acid (GLA) in Rhizomucor pusillus AUMC 11616.A and Mucor circinelloides AUMC 6696.A isolated from unusual habitats. Grown for 4 days at 28°C, shaking at 150 rpm, the maximum fungal biomass for AUMC 6696.A was 14.6 ± 0.2 g/L with arginine and 13.68 ± 0.1 g/L with asparagine, when these amino acids were used as single nitrogen sources, while AUMC 11616.A maximum biomass was 10.73 ± 0.8 g/L with glycine and 9.44 ± 0.6 g/L with valine. These were significantly higher than the ammonium nitrate control (p < 0.05). The highest levels of TFA were achieved with glycine for AUMC 11616.A, 26.2 ± 0.8% w/w of cell dry weight, and glutamic acid for AUMC 6696.A, 23.1 ± 1.3%. The highest GLA yield was seen with proline for AUMC 11616.A, 13.4 ± 0.6% w/w of TFA, and tryptophan for AUMC 6696.A, 12.8 ± 0.3%, which were 38% and 25% higher than the ammonium tartrate control. The effects of environmental factors such as temperature, pH, fermentation time, and agitation speed on biomass, total lipids, TFA, and GLA concentration of the target strains have also been investigated. Our results demonstrated that nitrogen assimilation through amino acid metabolism, as well as the use of glucose as a carbon source and abiotic factors, are integral to increasing the oleaginicity of tested strains. Few studies have addressed the role of amino acids in fermentation media, and this study sheds light on R. pusillus and M. circinelloides as promising candidates for the potential applications of amino acids as nitrogen sources in the production of lipids.

Abstract Trichoderma harzianum is one of the most commonly used fungal species used in the biological control of plant pathogenic fungi. Its ability to control biological pathogens is strengthened by the production of analytic enzymes. In the identification of these fungal species, it has been noted that morphological characteristics alone are insufficient. Hence, the aim of this study was to identify 12 Trichoderma isolates based on their molecular markers and to evaluate the antagonistic activity of these Trichoderma isolates against several plant pathogens. The 12 Trichoderma/Hypocrea isolates were harvested from the rhizosphere of healthy tomato plants in Abha region, Saudi Arabia. These isolates were identified by both morphological and molecular techniques, such as the sequencing of the 5.8S-ITS region. Furthermore, based on the taxonomy and phylogeny of these isolates, the genetic similarities based on the inter-simple sequence repeat-PCR markers separated the 12 Trichoderma isolates into 2 major clusters. The 12 isolates were then used in confrontational assays with Rhizoctonia solani, Pythium ultimum, and Alternaria solani. Most of the Trichoderma isolates showed a strong antagonistic activity, which could also be observed with the use of SEM. This study highlights the possibility of using T. harzianum in IPM programs as an effective biological agent against several pathogenic fungi.

Background and aim: Gastric Cancer (GC) is a leading cause of morbidity and mortality worldwide, particularly in developing nations, only a few suitable gastric cancer serum biomarkers with acceptable sensitivity and specificity exist. This work aims to highlight and uncover miR-30a-5p and miR-182–5p′s diagnostic roles regarding gastric cancer and their roles in predicting prognosis. Methods: 148 patients participated in this study. Groups I, II, and III had 47 patients with GC, 54 patients with benign gastric lesions, and 47 apparently healthy subjects of coincided age and gender as controls, respectively. All participants were clinically evaluated and subjected to CBC, serum CEA, and CA19-9 by ELISA, and real-time PCR tests of miR-30a-5p and miR-182–5p. Results: MiR30a-5p and miR-182–5p were down regulated in gastric cancer patients in Group I more than Groups II and III (P

Chamomile (Matricariarecutita L.) is one of the most important medicinal plants with various applications. The flowers and flower heads are the main organs inthe production of essential oil. The essential improvement goals of chamomile are considered to be high flower yield and oil content, as well asthe suitability for mechanical harvesting. The present study aimed to improve the flower yield, oil content and mechanical harvestability of German chamomile via chemical and physical mutagens. Three German chamomile populations (Fayum, Benysuif and Menia) were irradiated with 100, 200, 300 and 400 Gray doses of gamma rays, as well as chemically mutagenized using 0.001, 0.002 and 0.003 mol/mL of sodium azide for 4 h. The two mutagens produced a wide range of changes in the flowers’ shape and size. At M3 generation, 18 mutants (11 from gamma irradiation and 7 from sodium azide mutagenization) were selected and morphologically characterized. Five out of eighteen mutants were selected for morphological and chemical characterization for oil content, oil composition and oil quality in M4 generation. Two promising mutants, F/LF5-2-1 and B/HNOF 8-4-2, were selected based on their performance in most studied traits during three generations, as well as the high percentage of cut efficiency and a homogenous flower horizon, which qualify them as suitable candidates for mechanical harvesting. The two mutants are late flowering elite mutants; the F/LF5-2-1 mutant possessed the highest oil content (1.77%) and number of flowers/plant (1595), while the second promising B/HNOF 8-4-2 mutant hada high oil content (1.29%) and chamazulene percentage (13.98%) compared to control plants. These results suggest that the B/HNOF 8-4-2 and F/LF5-2-1 mutants could be integrated as potential parents into breeding programs for a high number of flowers, high oil content, oil composition and oil color traits for German chamomile improvement.

Diabetes mellitus is one of the most chronic metabolic diseases. In the past few years, our research group has synthesized and evaluated libraries of heterocyclic analogs against α-glucosidase and α-amylase enzymes and found encouraging results. The current study comprises the evaluation of benzimidazole-bearing thiosemicarbazone as antidiabetic agents. A library of fifteen derivatives (7–21) was synthesized, characterized via different spectroscopic techniques such as HREI-MS, NMR, and screened against α-glucosidase and α-amylase enzymes. All derivatives exhibited excellent to good biological inhibitory potentials. Derivatives 19 (IC50 = 1.30 ± 0.20 µM and 1.20 ± 0.20 µM) and 20 (IC50 = 1.60 ± 0.20 µM and 1.10 ± 0.01 µM) were found to be the most potent among the series when compared with standard drug acarbose (IC50 = 11.29 ± 0.07 and 11.12 ± 0.15 µM, respectively). These derivatives may potentially serve as the lead candidates for the development of new therapeutic representatives. The structure–activity relationship was carried out for all molecules which are mainly based upon the pattern of substituent/s on phenyl rings. Moreover, in silico docking studies were carried out to investigate the active binding mode of selected derivatives with the target enzymes.

Endophytic fungi including black aspergilli have the potential to synthesize multiple bioactive secondary metabolites. Therefore, the search for active metabolites from endophytic fungi against pathogenic microbes has become a necessity for alternative and promising strategies. In this study, 25 endophytic fungal isolates associated with Malus domestica were isolated, grown, and fermented on a solid rice medium. Subsequently, their ethyl acetate crude extracts were pretested for biological activity. One endophytic fungal isolate demonstrated the highest activity and was chosen for further investigation. Based on its phenotypic, ITS ribosomal gene sequences, and phylogenetic characterization, this isolate was identified as Aspergillus tubingensis strain AN103 with the accession number (KR184138). Chemical investigations of its fermented cultures yielded four compounds: Pyranonigrin A (1), Fonsecin (2), TMC 256 A1 (3), and Asperazine (4). Furthermore, 1H-NMR, HPLC, and LC-MS were performed for the identification and structure elucidation of these metabolites. The isolated pure compounds showed moderate-to-potent antibacterial activities against Pseudomonas aeruginosa and Escherichia coli (MIC value ranged from 31 and 121 to 14.5 and 58.3 μg/mL), respectively; in addition, the time–kill kinetics for the highly sensitive bacteria against isolated compounds was also investigated. The antifungal activity results show that (3) and (4) had the maximum effect against Fusarium solani and A. niger with inhibition zones of 16.40 ± 0.55 and 16.20 ± 0.20 mm, respectively, and (2) had the best effect against Candida albicans, with an inhibition zone of 17.8 ± 1.35 mm. Moreover, in a cytotoxicity assay against mouse lymphoma cell line L5178Y, (4) exhibited moderate cytotoxicity (49% inhibition), whereas (1–3) reported weak cytotoxicity (15, 26, and 19% inhibition), respectively. Our results reveal that these compounds might be useful to develop potential cytotoxic and antimicrobial drugs and an alternative source for various medical and pharmaceutical fields.

Determining the appropriate parents for breeding programs is the most important decision that plant breeders must make to maximize the genetic variability and produce excellent recombinant genotypes. Several methods are used to identify genotypes with desirable phenotypic features for breeding experiments. In this study, five kalanchoe genotypes were morphologically characterized by assessing plant height, number of inflorescences, number of flowers, flower length, flower diameter and number of petals. The analysis showed the distinction of yellow kalanchoe in the plant height trait, while the orange kalanchoe was distinguished in the number of inflorescences, the number of flowers and flower length traits, whereas the violet kalanchoe possessed the largest flower diameter and the highest number of petals. The molecular profiling was performed by random amplified polymorphism DNA (RAPD), inter-simple sequence repeats (ISSR) and start codon targeted (SCoT)-polymerase chain reaction (PCR) tools. Genomic DNA was extracted from young leaves and the PCR reactions were performed using ten primers for each SCoT, ISSR and RAPD marker. Only four out of ten primers showed amplicon profiles in all PCR markers. A total of 70 bands were generated by SCoT, ISSR and RAPD-PCR with 35 polymorphic bands and 35 monomorphic bands. The total number of bands of RAPD, ISSR and SCoT was 15, 17 and 38, respectively. The polymorphism percentages achieved by RAPD, ISSR and SCoT were 60.25%, 15% and 57%, respectively. The cluster analysis based on morphological data revealed two clusters. Cluster I consisted of violet and orange kalanchoe, and cluster II comprised red, yellow and purple kalanchoe. Whereas the cluster analysis based on molecular data revealed three clusters. Cluster I included only yellow kalanchoe, cluster II comprised orange and violet kalanchoe while cluster III comprised red, and purple kalanchoe. The study concluded that orange, violet and yellow kalanchoe are distinguished parents for breeding economically valued traits in kalanchoe. Also, the study concluded that SCoT and RAPD markers reproduced reliable banding patterns to assess the genetic polymorphism among kalanchoe genotypes that consider the basis stone for genetic improvements in ornamental plants.

Salinity is one of the harsh environmental stresses that destructively impact potato growth and production, particularly in arid regions. Exogenously applied safe–efficient materials is a vital approach for ameliorating plant growth, productivity, and quality under salinity stress. This study aimed at investigating the impact of foliar spray using folic acid (FA), ascorbic acid (AA), and salicylic acid (SA) at different concentrations (100, 150, or 200 mg/L) on plant growth, physiochemical ingredients, antioxidant defense system, tuber yield, and quality of potato (Solanum tuberosum L cv. Spunta) grown in salt-affected soil (EC = 7.14 dS/m) during two growing seasons. The exogenously applied antioxidant materials (FA, AA, and SA) significantly enhanced growth attributes (plant height, shoot fresh and dry weight, and leaves area), photosynthetic pigments (chlorophyll a and b and carotenoids), gas exchange (net photosynthetic rate, Pn; transpiration rate, Tr; and stomatal conductance, gs), nutrient content (N, P, and K), K+/ Na+ ratio, nonenzymatic antioxidant compounds (proline and soluble sugar content), enzymatic antioxidants (catalase (CAT), peroxidase (POX), superoxide dismutase (SOD), and ascorbate peroxidase (APX)) tuber yield traits, and tuber quality (dry matter, protein, starch percentage, total carbohydrates, and sugars percentage) compared with untreated plants in both seasons. Otherwise, exogenous application significantly decreased Na+ and Cl− compared to the untreated control under salt stress conditions. Among the assessed treatments, the applied foliar of AA at a rate of 200 mg/L was more effective in promoting salt tolerance, which can be employed in reducing the losses caused by salinity stress in potato grown in salt-affected soils.

The application of green nanotechnology in agriculture has been receiving substantial attention, especially in the development of new nano-fertilizers and nano-insecticides. Herein, the metabolites secreted by the fungal strain Penicillium chrysogenum are used as a reducing agent for selenium ions to form selenium nanoparticles (Se-NPs). The synthesized Se-NPs were characterized using color change, UV-Vis spectroscopy, Fourier transform infrared (FT-IR) spectroscopy, transmission electron microscopy (TEM), energy dispersive X-ray (EDX), X-ray diffraction (XRD), and dynamic light scattering (DLS). The biomass filtrate of the fungal strain changed from colorless to a ruby red color after mixing with sodium selenite with a maximum surface plasmon resonance at 262 nm. Data exhibits the successful formation of spherical, amorphous Se-NPs with sizes ranging between 3–15 nm and a weight percentage of 38.52%. The efficacy of Se-NPs on the growth performance of sunflower (Helianthus annuus L.) and inhibition of cutworm Agrotis ipsilon was investigated. The field experiment revealed the potentiality of Se-NPs to enhance the growth parameters and carotenoid content in sunflower, especially at 20 ppm. The chlorophylls, carbohydrates, proteins, phenolic compounds, and free proline contents were markedly promoted in response to Se-NPs concentrations. The antioxidant enzymes (peroxidase, catalase, superoxide dismutase, and polyphenol oxidase) were significantly decreased compared with the control. Data analysis showed that the highest mortality for the 1st, 2nd, 3rd, 4th, and 5th instar larvae of Agrotis ipsilon was achieved at 25 ppm with percentages of 89.7 ± 0.3, 78.3 ± 0.3, 72.3 ± 0.6, 63.7 ± 0.3, and 68.7 ± 0.3 respectively after 72 h.

Sesame is sensitive to waterlogging, and its growth is devastatingly impacted under excess moisture conditions. Thus, waterlogging tolerance is crucial to alleviate yield constraints, particularly under expected climate change. In this study, 119 diverse sesame genotypes were screened for their tolerance to 12, 24, 48, and 72 h of waterlogging relative to non-waterlogged conditions. All plants died under 72 h of waterlogging, while 13.45%, 31.93%, and 45.38% of genotypes survived at 48, 24, and 12 h, respectively. Based on the seedling parameters and waterlogging tolerance coefficients, genotypes BD-7008 and BD-6985 exhibited the highest tolerance to waterlogging, while BD-6996 and JP-01811 were the most sensitive ones. The responses of these four genotypes to waterlogged conditions were assessed at different plant growth stages—30, 40, and 50 days after sowing (DAS)—versus normal conditions. Waterlogging, particularly when it occurred within 30 DAS, destructively affected the physiological and morphological characteristics, which was reflected in the growth and yield attributes. Genotype BD-7008, followed by BD-6985, exhibited the highest chlorophyll and proline contents as well as enzymatic antioxidant activities, including superoxide dismutase (SOD), peroxidase (POD), and catalase (CAT). These biochemical and physiological adjustments ameliorated the adverse effects of waterlogging, resulting in higher yields for both genotypes. Conversely, JP-01811 presented the lowest chlorophyll and proline contents as well as enzymatic antioxidant activities, resulting in the poorest growth and seed yield.

In this study, we investigated aqueous extracts of three edible mushrooms: Agaricus bisporus (white button mushroom), Pleurotus columbinus (oyster mushroom), and Pleurotus sajor-caju (grey oyster mushroom). The extracts were biochemically characterized for total carbohydrate, phenolic, flavonoid, vitamin, and protein contents besides amino acid analysis. Triple TOF proteome analysis showed 30.1% similarity between proteomes of the two Pleurotus spp. All three extracts showed promising antiviral activities. While Pleurotus columbinus extract showed potent activity against adenovirus (Ad7, selectivity index (SI) = 4.2), Agaricus bisporus showed strong activity against herpes simplex II (HSV-2; SI = 3.7). The extracts showed low cytotoxicity against normal human peripheral blood mononuclear cells (PBMCs) and moderate cytotoxicity against prostate (PC3, DU-145); colorectal (Colo-205); cecum carcinoma (LS-513); liver carcinoma (HepG2); cervical cancer (HeLa); breast adenocarcinoma (MDA-MB-231 and MCF-7) as well as leukemia (CCRF-CEM); acute monocytic leukemia (THP1); acute promyelocytic leukemia (NB4); and lymphoma (U937) cell lines. Antioxidant activity was evaluated using 2,2-diphenyl-1-picryl-hydrazyl-hydrate (DPPH) radical scavenging, 2,2′-Azinobis-(3-Ethylbenzthiazolin-6-Sulfonic Acid) ABTS radical cation scavenging, and oxygen radical absorbance capacity (ORAC) assays. The three extracts showed potential antioxidant activities with the maximum activity recorded for Pleurotus columbinus (IC50 µg/mL) = 35.13 ± 3.27 for DPPH, 13.97 ± 4.91 for ABTS, and 29.42 ± 3.21 for ORAC assays.

The unique properties and advantages of edible films over conventional food packaging have led the way to their extensive exploration in recent years. Moreover, the incorporation of bioactive components during their production has further enhanced the intrinsic features of packaging materials. This study was aimed to develop edible and bioactive food packaging films comprising yeast incorporated into bacterial cellulose (BC) in conjunction with carboxymethyl cellulose (CMC) and glycerol (Gly) to extend the shelf life of packaged food materials. First, yeast biomass and BC hydrogels were produced by Meyerozyma guilliermondii (MT502203.1) and Gluconacetobacter xylinus (ATCC53582), respectively, and then the films were developed ex situ by mixing 30 wt.% CMC, 30 wt.% Gly, 2 wt.% yeast dry biomass, and 2 wt.% BC slurry. FE-SEM observation showed the successful incorporation of Gly and yeast into the fibrous cellulose matrix. FTIR spectroscopy confirmed the development of composite films through chemical interaction between BC, CMC, Gly, and yeast. The developed BC/CMC/Gly/yeast composite films showed high water solubility (42.86%). The yeast-incorporated films showed antimicrobial activities against three microbial strains, including Escherichia coli, Pseudomonas aeruginosa, and Saccharomyces aureus, by producing clear inhibition zones of 16 mm, 10 mm, and 15 mm, respectively, after 24 h. Moreover, the films were non-toxic against NIH-3T3 fibroblast cells. Finally, the coating of oranges and tomatoes with BC/CMC/Gly/yeast composites enhanced the shelf life at different storage temperatures. The BC/CMC/Gly/yeast composite film-coated oranges and tomatoes demonstrated acceptable sensory features such as odor and color, not only at 6 °C but also at room temperature and further elevated temperatures at 30 °C and 40 °C for up to two weeks. The findings of this study indicate that the developed BC/CMC/Gly/yeast composite films could be used as edible packaging material with high nutritional value and distinctive properties related to the film component, which would provide protection to foods and extend their shelf life, and thus could find applications in the food industry.

In this study, we examined aqueous extracts of the edible mushrooms Pleurotus ostreatus (oyster mushroom) and Lentinula edodes (shiitake mushroom). Proteome analysis was conducted using LC-Triple TOF-MS and showed the expression of 753 proteins by Pleurotus ostreatus, and 432 proteins by Lentinula edodes. Bioactive peptides: Rab GDP dissociation inhibitor, superoxide dismutase, thioredoxin reductase, serine proteinase and lectin, were identified in both mushrooms. The extracts also included promising bioactive compounds including phenolics, flavonoids, vitamins and amino acids. The extracts showed promising antiviral activities, with a selectivity index (SI) of 4.5 for Pleurotus ostreatus against adenovirus (Ad7), and a slight activity for Lentinula edodes against herpes simplex-II (HSV-2). The extracts were not cytotoxic to normal human peripheral blood mononuclear cells (PBMCs). On the contrary, they showed moderate cytotoxicity against various cancer cell lines. Additionally, antioxidant activity was assessed using DPPH radical scavenging, ABTS radical cation scavenging and ORAC assays. The two extracts showed potential antioxidant activities, with the maximum activity seen for Pleurotus ostreatus (IC50 µg/mL) = 39.46 ± 1.27 for DPPH; 11.22 ± 1.81 for ABTS; and 21.40 ± 2.20 for ORAC assays. This study encourages the use of these mushrooms in medicine in the light of their low cytotoxicity on normal PBMCs vis à vis their antiviral, antitumor and antioxidant capabilities.

Water-deficit stress poses tremendous constraints to sustainable agriculture, particularly under abrupt climate change. Hence, it is crucial to find eco-friendly approaches to ameliorate drought tolerance, especially for sensitive crops such as maize. This study aimed at assessing the impact of seed halo-priming on seedling vigor, grain yield, and water use efficiency of maize under various irrigation regimes. Laboratory trials evaluated the influence of seed halo-priming using two concentrations of sodium chloride solution, 4000 and 8000 ppm NaCl, versus unprimed seeds on seed germination and seedling vigor parameters. Field trials investigated the impact of halo-priming treatments on maize yield and water use efficiency (WUE) under four irrigation regimes comprising excessive (120% of estimated crop evapotranspiration, ETc), normal (100% ETc), and deficit (80 and 60% ETc) irrigation regimes. Over-irrigation by 20% did not produce significantly more grain yield but considerably reduced WUE. Deficit irrigation (80 and 60%ETc) gradually reduced grain yield and its attributes. Halo-priming treatments, particularly 4000 ppm NaCl, improved uniformity and germination speed, increased germination percentage and germination index, and produced more vigorous seedlings with heavier dry weight compared with unprimed seeds. Under field conditions, the plants originated from halo-primed seeds, especially with 4000 ppm NaCl, had higher grain yield and WUE compared with unprimed seeds under deficit irrigation regimes. The long-lasting stress memory induced by seed halo-priming, particularly with 4000 ppm NaCl, promoted maize seedling establishment, grain yield, and WUE and consequently mitigated the devastating impacts of drought stress.

Drought is one of the major threats to global food security. Biochar use in agriculture has received much attention and improving it through chemical modification offers a potential approach for enhancing crop productivity. There is still limited knowledge on how acidified biochar influences soil properties, and consequently its influences on the agricultural productivity of drought stressed plants. The water use efficiency (I-WUE) of drought stressed faba beans was investigated through the effects of acidified biochar (ACBio) (a 3:100 (w:w) combination of citric acid and biochar) on soil properties, growth, productivity, nutrient uptake, water productivity (WP), and irrigation. Two field experiments (2016/2017 and 2017/2018) were conducted in saline soil (ECe, 7.2 dS m−1) on faba been plants grown under three irrigation regimes (i.e., 100, 80, and 60% of crop evapotranspiration (ETc)) combined with three levels of ACBio (0, 5, and 10 t ha−1). Plants exposed to water stress presented a significant decrease in plant height, dry matter, leave area, chlorophyll content (SPAD), the quantum efficiency of photosystem II (Fv/Fm, Fv/F0, and PI), water status (membrane stability index and relative water content), and seed yield. Acidified biochar soil incorporation improved soil properties (chemical and physical), plant growth, physiological responses, WP, I-WUE, and contents of N, P, K, and Ca. Results revealed that the application of ACBio at 10 t ha−1 and 5 t ha−1 significantly increased seed yield by 38.7 and 25.8%, respectively, compared to the control. Therefore, ACBio incorporation may find application in the future as a potential soil amendment for improving growth and productivity of faba bean plants under deficit irrigation.

Salinity is a major abiotic stress that poses great obstacles to wheat production, especially in arid regions. The application of exogenous substances can enhance plant salt tolerance and increase its productivity under salinity stress. This work aimed to assess the mechanisms of selenium (Se) at different concentrations (2, 4 and 8 μM SeCl2) to mitigate hazardous impacts of salt toxicity at physiological, biochemical and agronomic levels in bread wheat. The results displayed that Se foliar application increased chlorophyll content, net photosynthetic rate, transpiration rate, stomatal conductance, relative water content, membrane stability index, excised leaf water retention, proline, total soluble sugars, Ca content, K content, antioxidant enzyme activities and non-enzymatic antioxidant compounds compared to untreated plants. On the other hand, Se application decreased the content of Na, hydrogen peroxide and superoxide contents. Accordingly, our findings recommend exogenous Se application (in particular 8 μM) to alleviate the deleterious effects induced by salinity stress and improve wheat yield attributes through enhancing antioxidant defense systems and photosynthetic capacity.

The metabolites of the fungal strain Rhizopus oryaze were used as a biocatalyst for the green-synthesis of magnesium oxide nanoparticles (MgO-NPs). The production methodology was optimized to attain the maximum productivity as follows: 4 mM of precursor, at pH 8, incubation temperature of 35 °C, and reaction time of 36 h between metabolites and precursor. The as-formed MgO-NPs were characterized by UV-Vis spectroscopy, TEM, SEM-EDX, XRD, DLS, FT-IR, and XPS analyses. These analytical techniques proved to gain crystalline, homogenous, and well-dispersed spherical MgO-NPs with an average size of 20.38 ± 9.9 nm. The potentiality of MgO-NPs was dose- and time-dependent. The biogenic MgO-NPs was found to be a promising antimicrobial agent against the pathogens including Staphylococcus aureus, Bacillus subtilis, Pseudomonas aeruginosa, Escherichia coli, and Candida albicans with inhibition zones of 10.6 ± 0.4, 11.5 ± 0.5, 13.7 ± 0.5, 14.3 ± 0.7, and 14.7 ± 0.6 mm, respectively, at 200 μg mL–1. Moreover, MgO-NPs manifested larvicidal and adult repellence activity against Culex pipiens at very low concentrations. The highest decolorization percentages of tanning effluents were 95.6 ± 1.6% at 100 µg/ 100 mL after 180 min. At this condition, the physicochemical parameters of tannery effluents, including TSS, TDS, BOD, COD, and conductivity were reduced with percentages of 97.9%, 98.2%, 87.8%, 95.9%, and 97.3%, respectively. Moreover, the chromium ion was adsorbed with percentages of 98.2% at optimum experimental conditions.

The discovery of eco-friendly, rapid, and cost-effective compounds to control diseases caused by microbes and insects are the main challenges. Herein, the magnesium oxide nanoparticles (MgO-NPs) are successfully fabricated by harnessing the metabolites secreted by Penicillium chrysogenum. The fabricated MgO-NPs were characterized using UV-Vis, XRD, TEM, DLS, EDX, FT-IR, and XPS analyses. Data showed the successful formation of crystallographic, spherical, well-dispersed MgO-NPs with sizes of 7–40 nm at a maximum wavelength of 250 nm. The EDX analysis confirms the presence of Mg and O ions as the main components with weight percentages of 13.62% and 7.76%, respectively. The activity of MgO-NPs as an antimicrobial agent was investigated against pathogens Staphylococcus aureus, Bacillus subtilis, Pseudomonas aeruginosa, Escherichia coli, and Candida albicans, and exhibited zone of inhibitions of 12.0 ± 0.0, 12.7 ± 0.9, 23.3 ± 0.8, 17.7 ± 1.6, and 14.7 ± 0.6 mm respectively, at 200 µg mL−1. The activity is decreased by decreasing the MgO-NPs concentration. The biogenic MgO-NPs exhibit high efficacy against different larvae instar and pupa of Anopheles stephensi, with LC50 values of 12.5–15.5 ppm for I–IV larvae instar and 16.5 ppm for the pupa. Additionally, 5 mg/cm2 of MgO-NPs showed the highest protection percentages against adults of Anopheles stephensi, with values of 100% for 150 min and 67.6% ± 1.4% for 210 min.

Mastitis is a significant disease affecting dairy cattle farms in Egypt. The current study aimed to investigate the prevalence and major bacterial pathogens causing subclinical mastitis (SCM) in three bovine dairy herds, with a history of SCM, at three Governorates in North Upper Egypt. The antimicrobial resistance profiles and specific virulence-associated genes causing bovine SCM were investigated. One thousand sixty-quarter milk samples (QMS) were collected aseptically from 270 apparently healthy cows in three farms and examined. The total prevalence of SCM was 46% and 44.8% based on California Mastitis Test (CMT) and Somatic Cell Count (SCC), respectively. Bacteriological examination of CMT positive quarters revealed that the prevalence of bacterial isolation in subclinically mastitic quarters was 90.4% (26 and 64.3% had single and mixed isolates, respectively). The most frequent bacterial isolates were E. coli (49.8%), Staphylococcus aureus (44.9%), streptococci (44.1%) and non-aureus staphylococci (NAS) (37.1%). Antimicrobial susceptibility testing of isolates revealed a high degree of resistance to the most commonly used antimicrobial compound in human and veterinary medicine. Implementation of PCR revealed the presence of mecA and blaZ genes in 60% and 46.7% of S. aureus isolates and in 26.7% and 53.3% of NAS, respectively. Meanwhile 73.3% of streptococci isolates harbored aph(3’)-IIIa gene conferring resistance to aminoglycosides and cfb gene. All E. coli isolates harbored tetA gene conferring resistance to tetracycline and sul1 gene conferring resistance to sulfonamides. The fimH and tsh genes were found in 80% and 60%, respectively. A significant association between the phenotypes and genotypes of AMR in different bacteria was recorded. The presence of a high prevalence of SCM in dairy animals impacts milk production and milk quality. The coexistence of pathogenic bacteria in milk is alarming, threatens human health and has a public health significance. Herd health improvement interventions are required to protect human health and society.

Dual-purpose barley is an alternative approach to producing high-quality forage yield plus an acceptable grain yield in marginal environments of arid regions that are characterized by lack of forage. Field experiment was performed in two consecutive growing seasons at an arid region affected by salinity in irrigation water and soil at Western Sinai Peninsula in Egypt. The study aimed to optimize sowing date and screen salt-tolerant barley genotypes that perform better in terms of forage yield and quality as well as grain and biomass yield production in salt-affected environment. Sowing dates, genotypes, and their interaction significantly impacted most of the studied variables such as forage yield, crude protein yield, and grain and biomass yields. The early sowing in late October yielded higher than intermediate sowing in mid-November and late sowing in early December. Some of the tested genotypes performed better than others as indicated by about 50% higher forage yield, 6% crude protein content, 39% grain and 21% biological yields (total aboveground dry matter), suggesting higher adaptation capacity. Interestingly, grain and biological yields did not differ significantly between dual-purpose approach and grain-only pattern. In conclusion, dual-purpose barley was found favorable for producing grain and forage production in similar environments under early sowing date.

Water shortage is a major environmental stress that destructively impacts maize production, particularly in arid regions. Therefore, improving irrigation management and increasing productivity per unit of water applied are needed, especially under the rising temperature and precipitation fluctuations induced by climate change. Laboratory and field trials were carried out in the present study, which were aimed at assessing the possibility of promoting maize germination, growth, grain yield and crop water productivity (CWP) using seed priming under different irrigation regimes. Two seed priming treatments, i.e., hydro-priming and hardening versus unprimed seeds, were applied under four irrigation regimes, i.e., 120, 100, 80 and 60% of estimated crop evapotranspiration (ETc). The obtained results indicated that increasing irrigation water from 100% up to 120% ETc did not significantly increase grain yield or contributing traits, while it decreased CWP. Deficit irrigation of 80 and 60% ETc gradually decreased grain yield and all attributed traits. Seed priming significantly ameliorated seedlings’ vigor as indicated by earlier germination, higher germination percentage, longer roots and shoots, and heavier fresh and dry weight than unprimed seeds with the superiority of hardening treatment. Additionally, under field conditions, seed priming significantly increased grain yield, yield contributing traits and CWP compared with unprimed treatment. Interestingly, the results reflect the role of seed priming, particularly hardening, in mitigating negative impacts of drought stress and enhancing maize growth, grain yield and attributed traits as well as CWP under deficit irrigation conditions. This was demonstrated by a significant increase in grain yield and CWP under moderate drought and severe drought conditions compared with unprimed treatment. These results highlight that efficient irrigation management and seed priming can increase maize yield and water productivity in arid environments.

The current rise of multidrug-resistant (MDR) Gram-negative Enterobacteriaceae including the extended-spectrum β-lactamase (ESBL)-producing organisms and carbapenem-resistant Enterobacteriaceae (CRE) has been increasingly reported worldwide, posing new challenges to health care facilities. Accordingly, we evaluated the impact of multimodal infection control interventions at one of the major tertiary healthcare settings in Egypt for the aim of combating infections by the respective pathogens. During the 6-month pre-intervention period, the incidence rate of CRE and ESBL-producing clinical cultures were 1.3 and 0.8/1000 patient days, respectively. During the post-intervention period, the incidence of CRE and ESBL producers continued to decrease, reaching 0.5 and 0.28/1000 patient days, respectively. The susceptibility rate to carbapenems among ESBL producers ranged from 91.4% (ertapenem) to 98.3% (imipenem), amikacin (93%), gentamicin (56.9%), and tobramycin (46.6%). CRE showed the highest resistance pattern toward all of the tested β-lactams and aminoglycosides, ranging from 87.3% to 94.5%. Both CRE and ESBL producers showed a high susceptibility rate (greater than 85.5%) to colistin and tigecycline. In conclusion, our findings revealed the effectiveness of implementing multidisciplinary approaches in controlling and treating infections elicited by CRE and ESBL producers.

Twenty-one fungal strains were isolated from dye-contaminated soil; out of them, two fungal strains A2 and G2-1 showed the highest decolorization capacity for real textile effluent and were, hence, identified as Aspergillus flavus and Fusarium oxysporium based on morphological and molecular methods. The highest decolorization percentage of 78.12 ± 2.1% was attained in the biotreatment with fungal consortium followed by A. flavus and F. oxysporium separately with removal percentages of 54.68 ± 1.2% and 52.41 ± 1.0%, respectively. Additionally, ultraviolet-visible spectroscopy of the treated effluent showed that a maximum peak (λmax) of 415 nm was reduced as compared with the control. The indicators of wastewater treatment efficacy, namely total dissolved solids, total suspended solids, conductivity, biological oxygen demand, and chemical oxygen demand with removal percentages of 78.2, 78.4, 58.2, 78.1, and 77.6%, respectively, demonstrated a considerable decrease in values due to fungal consortium treatment. The reduction in peak and mass area along with the appearance of new peaks in GC-MS confirms a successful biodegradation process. The toxicity of treated textile effluents on the seed germination of Vicia faba was decreased as compared with the control. The shoot length after irrigation with effluents treated by the fungal consortium was 15.12 ± 1.01 cm as compared with that treated by tap-water, which was 17.8 ± 0.7 cm. Finally, we recommended the decrease of excessive uses of synthetic dyes and utilized biological approaches for the treatment of real textile effluents to reuse in irrigation of uneaten plants especially with water scarcity worldwide.

Herein, CuO-NPs were fabricated by harnessing metabolites of Aspergillus niger strain (G3-1) and characterized using UV–vis spectroscopy, XRD, TEM, SEM-EDX, FT-IR, and XPS. Spherical, crystallographic CuO-NPs were synthesized in sizes ranging from 14.0 to 47.4 nm, as indicated by TEM and XRD. EDX and XPS confirmed the presence of Cu and O with weight percentages of 62.96% and 22.93%, respectively, at varied bending energies. FT-IR spectra identified functional groups of metabolites that could act as reducing, capping, and stabilizing agents to the CuO-NPs. The insecticidal activity of CuO-NPs against wheat grain insects Sitophilus granarius and Rhyzopertha dominica was dose- and time-dependent. The mortality percentages due to NP treatment were 55–94.4% (S. granarius) and 70–90% (R. dominica). A botanical experiment was done in a randomized block design. Low CuO-NP concentration (50 ppm) caused significant increases in growth characteristics (shoot and root length, fresh and dry weight of shoot and root, and leaves number), photosynthetic pigments (total chlorophylls and carotenoids), and antioxidant enzymes of wheat plants. There was no significant change in carbohydrate or protein content. The use of CuO-NPs is a promising tool to control grain insects and enhance wheat growth performance.

Salinity in soil or irrigation water requires developing genetically salt-tolerant genotypes, especially in arid regions. Developing salt-tolerant and high-yielding wheat genotypes has become more urgent in particular with continuing global population growth and abrupt climate changes. The current study aimed at investigating the genetic variability of new breeding lines in three advanced generations F6–F8 under salinity stress. The evaluated advanced lines were derived through accurate pedigree selection under actual saline field conditions (7.74 dS/m) and using saline water in irrigation (8.35 dS/m). Ninety-four F6 lines were evaluated in 2017–2018 and reduced by selection to thirty-seven F7 lines in 2018–2019 and afterward to thirty-four F8 lines in 2019–2020 based on grain yield and related traits compared with adopted check cultivars. Significant genetic variability was detected for all evaluated agronomic traits across generations in the salt-stressed field. The elite F8 breeding lines displayed higher performance than the adopted check cultivars. These lines were classified based on yield index into four groups using hierarchical clustering ranging from highly salt-tolerant to slightly salt-tolerant genotypes, which efficiently enhance the narrow genetic pool of salt-tolerance. The detected response to selection and high to intermediate broad-sense heritability for measured traits displayed their potentiality to be utilized through advanced generations under salinity stress for identifying salt-tolerant breeding lines.