An Ethnobotanical, Phytochemical Analysis, Antimicrobial and Biological Studies of Pulicaria crispa as a Graze Promising Shrub.

Due to the global issue of antimicrobial resistance, one of the most significant challenges in microbiological research is to develop a replacement antibiotic with minimal adverse effects. The wild shrub Pulicaria crispa (gethgath) has been traditionally used for camel and ruminant grazing. While prior research has demonstrated its antimicrobial properties against human diseases, no investigations have been conducted on its efficacy against animal pathogens. The objective of this study is to explore the ethnobotanical, phytochemical, antioxidant, anticancer, and antimicrobial activity of Pulicaria crispa aqueous and solvent extracts against a range of standard and animal pathogens. All of the extracts demonstrated antimicrobial, antioxidant, and anticancer properties, containing bioactive compounds. Notably, the ethyl acetate extract of P. crispa exhibited the strongest antimicrobial activity against tested Gram-positive and Gram-negative bacteria and fungi. The chloroform fraction displayed the highest antioxidant activity. Additionally, the ethyl acetate fraction showed promising anticancer activity against breast (MCF-7) and lung (A549) cancer cells. These findings confirm that Pulicaria crispa is a valuable shrub with potential applications as a natural alternative for antimicrobial, antioxidant, and anticancer treatments in both human and veterinary medicine.

Assessment of Foodborne Bacterial Pathogens in Buffalo Raw Milk Using Polymerase Chain Reaction Based Assay.

Milk is a putrescible commodity that is extremely prone to microbial contamination. Primarily, milk and dairy products are believed to be easily contaminated by pathogenic microorganisms, including Listeria monocytogenes, Salmonella spp., and Staphylococcus aureus. The microbiological quality of raw milk and dairy products regarding foodborne pathogens is of paramount importance due to concern of human health. In this study 400 buffalo raw milk samples were screened for assessing the prevalence of L. monocytogenes, Salmonella spp., and S. aureus. This study implemented uniplex-polymerase chain reaction (u-PCR) and multiplex-polymerase chain reaction (m-PCR) assays for the fast simultaneous detection of these pathogens comparing to the conventional culturing methods. Raw milk samples were found contaminated with the prevalence of 2.2%, 4.0%, and 14.2% for L. monocytogenes, Salmonella spp., and S. aureus, respectively. These pathogens were detected with the optimized polymerase chain reaction assays after 6 h of enrichment. u-PCR and m-PCR demonstrated the limit of detection as 104, 102, and 10 cells/mL after 6, 12, 18, and 24 h for each culture of the pathogens. A high sensitivity (10 colony-forming unit [CFU]/mL) of the m-PCR protocol was noted. The developed protocol is a cost-effective and rapid method for the simultaneous detection of pathogens associated with raw milk and dairy industries.

Elicitor-Induced Metabolomics Analysis of Halodule pinifolia Suspension Culture for an Alternative Antifungal Screening Approach against Candida albicans.

Elicitors are the agents that stimulate the defense responses of plants, and accumulate specialized metabolites in plant tissue culture. This study investigated the elicitor-feeding response of H. pinifolia suspension cell cultures (SCC) for metabolomics analysis and screening of specialized compounds against Candida albicans. Methyl jasmonate (MeJA) was used as an elicitor, and treatment of SCC at a concentration of 20 µM MeJA resulted in the maximum rosmarinic acid (RA) accumulation (117 mg/g dry weight), with transcript levels of RA biosynthetic genes HpPAL, HpC4H, and Hp4CL being 4.2, 2.5, and 3.7-fold higher, respectively, than the controls. GC-MS-based metabolomics analysis revealed a total of 47 metabolites, including 30 organic acids, six amino acids, six flavonoids, two sugars, two plant growth regulators, and one vitamin, which were significantly different between control and MeJA-treated cells. Furthermore, five phenolic acids were discovered at higher concentrations, including p-anisic acid, p-coumaric acid, caffeic acid, vanillic acid, and rosmarinic acid, and were purified and structurally elucidated for alternative antifungal screening against C. albicans and the evaluation of ADMET properties. The results from antifungal screening revealed that RA at MIC of 31.25 mg/L exhibited the lowest growth percentage of C. albicans (1.99%), with higher inhibition of isocitrate lyase 1 (ICL 1) enzyme (93.1%), followed by p-anisic acid (86.2%) and caffeic acid (85.1%), respectively. The drug likeliness and ADMET properties of RA exhibited promising results, with a bioactivity score of 0.57, 0.15, and 0.24 for nuclear receptor ligand, protease inhibitor, and enzyme inhibitor, respectively. Therefore, MeJA appears to have a significant effect on enhanced RA accumulation in H. pinifoia cells with phenylpropanoid transcript expression, and acts as an ICL1 inhibitor of C. albicans.

Ethnobotanical, phytochemical and antimicrobial activity of Halexylon salicornicum (Ramth) as a graze and promising shrub against selected animal microbes.

Folk medicine, including plants, has been utilized for humans and animals since the dawn of civilization. Because of the widespread problem of antimicrobial resistance around the world, one of the mainly significant challenges in microbiological research is to achieve a replacement antibiotic with the fewest adverse effects. Camel and ruminant grazing were provided by the wild shrub Halexylon salicornicum (Ramth). However, despite prior research demonstrating its antimicrobial action against human diseases, no investigations on its antimicrobial activity against animal pathogens have been conducted to far. The goal of this study is to investigate whether Halexylon salicornicum aqueous and solvent extracts have antimicrobial activity on a variety of animal pathogens isolated from cattle and poultry using the agar well diffusion method (Enterococcus faecium, Shigella flexneri, Bacillus Cereus, Klebsiella pneumoniae, Staphylococcus aureus, Escherichia coli, Streptococcus pyogens, Pseudomonas aerogenes, Salmonella typhimurium, and Candida albicans) moreover it's ethnobotanical and phytochemical. All of the extracts tested had antimicrobial efficacy against tested strains and included bioactive chemicals, particularly the acetone extract, had the highest antibacterial activity. As a result, it can be stated that Halexylon salicornicum is a promising important shrub that might be used as a natural antimicrobial alternative for animals or as a food additive.

Transcriptome Profiling Reveals Differential Gene Expression of Laccase Genes in Aspergillus terreus KC462061 during Biodegradation of Crude Oil.

Fungal laccases have high catalytic efficiency and are utilized for the removal of crude oil because they oxidize various aliphatic and aromatic hydrocarbons and convert them into harmless compounds or less toxic compounds, thus accelerating the biodegradation potential of crude oil. Laccases are important gene families and the function of laccases genes varied widely based on transcription and function. Biodegradation of crude oil using Aspergillus terreus KC462061 was studied in the current study beside the transcription level of eight laccase (Lcc) genes have participated in biodegradation in the presence of aromatic compounds, and metal ions. Time-course profiles of laccase activity in the presence of crude oil indicated that the five inducers individual or combined have a very positive on laccase activity. In the status of the existence of crude oil, the synergistic effect of Cu-ABTS compound caused an increase in laccase yields up to 22-fold after 10 days than control. The biodegradation efficiencies of A. terreus KC462061 for aliphatic and aromatic hydrocarbons of crude oil were 82.1 ± 0.2% and 77.4 ± 0.6%, respectively. The crude oil biodegradation efficiency was improved by the supplemented Cu-ABTS compound in A. terreus KC462061. Gas chromatography-mass spectrometry was a very accurate tool to demonstrate the biodegradation efficiencies of A. terreus KC462061 for crude oil. Significant differences were observed in the SDS-PAGE of A. terreus KC462061 band intensities of laccase proteins after the addition of five inducers, but the Cu-ABTS compound highly affects very particular laccase electrophoresis. Quantitative real-time polymerase chain reaction (qPCR) was used for the analysis of transcription profile of eight laccase genes in A. terreus KC462061 with a verified reference gene. Cu2+ ions and Cu-ABTS were highly effective for efficient laccase expression profiling, mainly via Lcc11 and 12 transcription induction. The current study will explain the theoretical foundation for laccase transcription in A. terreus KC462061, paving the road for commercialization and usage.

Kinetic, Isotherm and Thermodynamic Aspects of Zn2+ Biosorption by Spirulina platensis: Optimization of Process Variables by Response Surface Methodology.

The aim of this study was to assess the efficiency of Spirulina platensis for removing Zn2+ ions from the aqueous solutions. The optimized conditions of 4.48 g/L algal dose, pH of 6.62 and initial zinc concentration of 29.72 mg/L obtained by response surface methodology were employed for Zn2+ biosorption by S. platensis and up to 97.90% Zn2+ was removed, showing that there is a favorable harmony between the experimental data and model predictions. Different kinetic and equilibrium models were used to characterize the biosorption manner of Spirulina as a biosorbent. The kinetic manner of Zn2+ biosorption was well characterized by the pseudo-second-order, implying that the adsorption process is chemical in nature. The Langmuir and Dubinin-Radushkevich isotherm models were best fit to the equilibrium data. The maximum adsorption capacity of the Langmuir monolayer was 50.7 mg/g. Furthermore, the thermodynamic analysis revealed that Zn2+ biosorption was endothermic, spontaneous and feasible. As a result of biosorption process, FTIR, SEM, and EDX investigations indicated noticeable alterations in the algal biomass's properties. Therefore, the dried Spirulina biomass has been shown to be cost-effective and efficient for removing the heavy metals, particularly zinc ions from wastewater, and the method is practicable, and environmentally acceptable.

Trichogenic Silver-Based Nanoparticles for Suppression of Fungi Involved in Damping-Off of Cotton Seedlings.

Mycogenic silver nanoparticles (AgNPs) produced by some biocontrol agents have shown the ability to inhibit the growth of numerous plant pathogenic fungi, which may be a unique method of disease management. This study describes the extracellular production of AgNPs by Trichoderma harzianum. The size, shape, charge, and composition of the AgNPs were subsequently studied by UV-visible spectroscopy, DLS, zeta potential, TEM, SEM, and EDX, among other methods. The AgNPs had sizes ranging from 6 to 15 nm. The antifungal activities of bio-synthesized AgNPs and two commercial fungicides (Moncut and Maxim XL) were tested against three soil-borne diseases (Fusarium fujikuroi, Rhizoctonia solani, and Macrophomina phaseolina). Cotton seedling illnesses were significantly reduced under greenhouse settings after significant in vitro antifungal activity was documented for the control of plant pathogenic fungi. The use of biocontrol agents such as T. harzianum, for example, may be a safe strategy for synthesizing AgNPs and using them to combat fungus in Egyptian cotton.

Trichoderma harzianum-Mediated ZnO Nanoparticles: A Green Tool for Controlling Soil-Borne Pathogens in Cotton.

ZnO-based nanomaterials have high antifungal effects, such as inhibition of growth and reproduction of some pathogenic fungi, such as Fusarium sp., Rhizoctonia solani and Macrophomina phaseolina. Therefore, we report the extracellular synthesis of ZnONPs using a potential fungal antagonist (Trichoderma harzianum). ZnONPs were then characterized for their size, shape, charge and composition by visual analysis, UV-visible spectrometry, X-ray diffraction (XRD), Zeta potential, transmission electron microscopy (TEM), scanning electron microscopy (SEM) and energy-dispersive X-ray analysis (EDX). The TEM test confirmed that the size of the produced ZnONPs was 8-23 nm. The green synthesized ZnONPs were characterized by Fourier transform infrared spectroscopy (FTIR) studies to reveal the functional group attributed to the formation of ZnONPs. For the first time, trichogenic ZnONPs were shown to have fungicidal action against three soil-cotton pathogenic fungi in the laboratory and greenhouse. An antifungal examination was used to evaluate the bioactivity of the mycogenic ZnONPs in addition to two chemical fungicides (Moncut and Maxim XL) against three soil-borne pathogens, including Fusarium sp., Rhizoctonia solani and Macrophomina phaseolina. The findings of this study show a novel fungicidal activity in in vitro assay for complete inhibition of fungal growth of tested plant pathogenic fungi, as well as a considerable reduction in cotton seedling disease symptoms under greenhouse conditions. The formulation of a trichogenic ZnONPs form was found to increase its antifungal effect significantly. Finally, the utilization of biocontrol agents, such as T. harzianum, could be a safe strategy for the synthesis of a medium-scale of ZnONPs and employ it for fungal disease control in cotton.

Efficacy of Acacia nilotica aqueous extract in treating biofilm-forming and multidrug resistant uropathogens isolated from patients with UTI syndrome.

Escherichia coli is the dominant bacterial cause of UTI among the uropathogens in both developed and developing countries. This study is to investigate the effect of Acacia nilotica aqueous extract on the survival and biofilm of isolated pathogens to reduce UTIs diseases. A total of 170 urine samples were collected from Luxor general hospital and private medical analysis laboratories in Luxor providence, Egypt. Samples were screened for the incidence of uropathogens by biochemical tests, antibiotics susceptibility, detection of virulence, and antibiotic-resistant genes by multiplex PCR, biofilm formation, and time-killing assay. Escherichia coli is by far the most prevalent causative agent with the percentage of 73.7% followed by Klebsiella pneumoniae, Proteus mirabilis, Pseudomonas aeuroginosa, and Acinetobacter baumanii. Isolates were multidrug-resistant containing blaTEM, blaSHV, blaCTX, qnrs, and aac(3)-Ia resistant genes. All isolates were sensitive to 15-16.7 mg ml-1 of Acacia nilotica aqueous extract. Time killing assay confirmed the bactericidal effect of the extract over time (20-24 h). A high percentage of 3-Cyclohexane-1-Carboxaldehyde, 2,6,6-trimethyl (23.5%); á-Selinene (15.12%); Oleic Acid (14.52%); Globulol (11.35%) were detected among 19 bioactive phytochemical compounds in the aqueous extract of A. nilotica over the GC-mass spectra analysis. The plant extract reduced significantly the biofilm activity of E. coli, K. pneumoniae, P. mirabilis, and P. aeuroginosa by 62.6, 59. 03, 48.9 and 39.2%, respectively. The challenge to improve the production of A. nilotica phytochemicals is considered a very low price for the return.

Phylogenetic Diversity of Trichoderma Strains and Their Antagonistic Potential against Soil-Borne Pathogens under Stress Conditions.

Trichoderma species are known as excellent biocontrol agents against soil-borne pathogens that cause considerable crop losses. Eight strains of Trichoderma were isolated from five Egyptian regions. They identified based on translation elongation factor-1α (TEF1) sequencing as four different Trichoderma species: Trichoderma asperellum, Trichoderma harzianum, Trichoderma viride, and Trichoderma longibrachiatum. Optimal growth conditions (temperature and media), and the phosphate solubilization capability of Trichoderma strains were evaluated in vitro. Further, the ability of these strains to antagonize Fusarium solani, Macrophomina phaseolina, and Fusarium graminearum was also evaluated. The results revealed that Trichoderma harzianum (Th6) exhibited the highest antagonistic ability against F. solani, M. phaseolina and F. graminearum with inhibition rates of 71.42%, 72.97%, and 84.61%, respectively. Trichoderma viride (Tv8) exhibited the lowest antagonism against the same pathogens with inhibition rates of 50%, 64% and 69.23%, respectively. Simple-sequence repeats (SSRs) and random amplified polymorphic DNA (RAPD) markers were used to evaluate the genetic variability of the Trichoderma strains. The results revealed that of 45 RAPD amplified bands, 36 bands (80%) were polymorphic and of SSRs amplified 36 bands, 31 bands (86.11%) were polymorphic. The amplification of calmodulin and ß-1,3-endoglucanase was noted at 500 bp and 230 bp, respectively. Data indicated that T. viride (Tv8) had the highest phosphate solubilization index (10.0 mm), while T. harzianum (Th6) had the lowest phosphate solubilization index (4.0 mm). In conclusion, T. harzianum (Th6) had the highest antagonistic activity in dual culture assay along with the growth rate; while T. viride (Tv8) had the highest phosphate solubilization activity. There are still gaps in obtaining new formulations, selecting potent Trichoderma strains to confirm disease control in planta. For improving Trichoderma recommendation in the organic agricultural system and sustaining the fertility of the soil, the field application of highly antagonistic biocontrol agents in different types of soil and plant species will be the first approach toward bio-pesticide treatments along with bio-fertilizer inoculation. Furthermore, secondary metabolites will be investigated for the most promising strains with the combination of different pathogens and application timing.