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The main focus of this study was to create a stable and efficient nanoemulsion (NE) using Callistemon citrinus essential oil (EO). Various factors affecting the NE's stability were optimized including oil %, Tween 80%, time of sonication, and its accelerated stability was examined. The research also considered the antibacterial, antifungal, and larvicidal effects of the optimized NE (B10). The optimum NE stood out for its stability, featuring a particle size of 33.15 ± 0.32 nm. Analysis via IR spectroscopy confirmed successful EO encapsulation in B10. The formulation remained stable for six months, with B10 showing significantly higher antibacterial and antifungal potency compared to the pure oil. When samples were subjected to tests against Fusarium oxysporum, B10 exhibited a MIC value of 62.5 mg/mL, whereas the pure oil showed a MIC value of 250 mg/mL. This indicates that the B10 formulation was 50 times more effective than the EO. In terms of antibacterial activity against Escherichia coli, the MIC value was 0.256 mg/mL for B10 and 4 mg/mL for the EO. Also, pure oil and B10 displayed larvicidal effects against Chilo suppressalis (Walker) larvae, with B10 eliminating 95.2% of larvae in 48 h. Overall, stable and optimum C. citrinus NE with its strong antimicrobial qualities, shows promise as an effective fungicide and insecticide.

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BACKGROUND: The hawthorn has recently been used as a popular herbal medicine in food applications and phytotherapy, especially for the cardiovascular system. METHODS: In this study, phytochemicals were evaluated by LC-ESI-MS, GC-MS, and biological activity, including antioxidant (DPPH test) and antibacterial (broth dilution assay), in different extracts of Crataegus pentagyna fruit, leaf, and root. RESULTS: Globally, 49 phenolics were tentatively identified using HPLC-ESI-MS/MS in the hydro-methanolic extract of the fruit (major apigenin, caffeoylquinic acid derivative, and 4-O-(3'-O-glucopyranosyl)-caffeoyl quinic acid), 42 in the leaf (major salicylic acid, naringenin-6-C-glucoside, and naringin), and 33 in the root (major naringenin-7-O-neohesperidoside, isovitexin-2″-O-rhamnoside, and 4-O-(3'-O-glucopyranosyl)-caffeoyl quinic acid). The major group compounds analyzed by GC-MS in petroleum ether extracts were hydrocarbons (63.80%) and fatty acids and their derivatives (11.77%) in fruit, hydrocarbons (49.20%) and fatty acids and their derivatives (13.85%) in leaf, and hydrocarbons (53.96%) and terpenes (13.06%) in root. All samples exhibited promising phytochemical profile (total phenol, flavonoid, phenolic acid, and anthocyanin), antioxidant and antibacterial capacities, especially in hydro-methanolic extract of fruit (210.22 ± 0.44 mg GAE/g DE; 79.93 ± 0.54 mg QE/g DE; 194.64 ± 0.32 mg CAE/g DE; 85.37 ± 0.13 mg cyanidin 3-glucoside/100 g FW; DPPH: 15.43 ± 0.65 µg/mL; MIC: 0.15-0.62 µg/mL; and MBC: 0.62-1.25 mg/mL), followed by the leaf and root extracts, respectively. The PCA and heatmap analysis results distinguished metabolite profile differences for samples. CONCLUSION: The results of the present work provide scientific support for C. pentagyna as antimicrobial agents and natural antioxidants in human health and food preservation.

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An efficient metal-free approach for the synthesis of N-coumaryl amino acids and the first one-step synthesis of 4-hydrazinocoumarin from 4-hydroxycoumarin was developed. The nucleophilic addition of amino acid methyl esters to 4-tosylcoumarins produced a series of 4-N-α-coumaryl amino acids in good to excellent yields without racemization. The antioxidant activities of the synthesized compounds were investigated using DPPH and FRAP methods. 4-Hydrazinocoumarin and N-coumaryl tyrosine had the best antioxidant activity. The antimicrobial activities of the compounds against Gram-positive was stronger than Gram-negative. 4-Hydrazinocoumarin showed the best antibacterial effect.

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Edible films containing essential oils (EO) as natural antibacterial agents are promising systems for food preservation. In this work, nanoemulsions of Thymus daenensis EO (wild; F1 and cultivated; F2) were loaded in hydroxyl propyl methyl cellulose (HPMC) films and the effect of different parameters (polymer, plasticizer, and EO concentration) on the film properties were analyzed and optimized. Prepared HPMC films were characterized in terms of EO loading, morphology, mechanical properties, and the antibacterial activity. The results of SEM showed uniform incorporation of nanoemulsions into the edible film. Investigation of the mechanical properties of two edible films revealed a plasticizing effect of T. daenensis EO on the films. Also, edible films had noticeable antimicrobial activity against selected microorganisms, i.e. 47.0±2.5mm and 22.6±0.5mm zone of inhibition against S. aureus for films containing F1 and F2, respectively. Incorporation of nanoemulsions into the HPMC films can be used for active food preservation.

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Using ultrasonic technology, trans-cinnamaldehyde as a natural antibacterial compound was used to prepare nano size emulsions to increase its bioavailability and therefore bactericidal action. Nanoemulsions containing trans-cinnamaldehyde as an active agent and 1,8 cineol as co additive oil (Ostwald ripening inhibitor) were formulated using probe sonicator. Three different determining factors, namely time of sonication, surfactant to oil ratio and type of emulsifier (Tween 80 and Tween 20) were investigated to enhance the stability profile. In addition, the effect of changes in the particle size and emulsifier on the antibacterial activity against Escherichia coli, Pseudomonas aeruginosa and Staphylococcus aureus were examined using agar dilution method. Then, the effect of optimized formulation on the membrane fluidity and cell constituent release, were investigated by analysis of membrane lipids using GC-MS and IR spectrometry, respectively. The data showed that a 15min sonication of the formulation containing Tween 80 as emulsifier with surfactant to oil ratio of 2:1 (w/w) resulted in a significant stability for 6months with considerably small particle size of 27.76±0.37nm. Furthermore, the nanoemulsion showed great antibacterial activity and could reduce the minimum inhibitory concentration (MIC) from 8 to 1mg/mL against E. coli and S. aureus, and from 16 to 2mg/mL against P. aeruginosa. Interestingly, E. coli's membrane fluidity increased dramatically after treatment with the optimum nanoemulsion (T804). This study revealed that nanoemulsion of trans-cinnamaldehyde and 1,8 cineol has substantial antibacterial activity against selected microorganisms.

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Natural preservatives are being extensively investigated for their potential industrial applications in foods and other products. In this work, an essential oil (Thymus daenensis) was formulated as a water-dispersible nanoemulsion (diameter=143nm) using high-intensity ultrasound. The antibacterial activity of the essential oil in both pure and nanoemulsion forms was measured against an important food-borne pathogen bacterium, Escherichia coli. Antibacterial activity was determined by measuring the minimum inhibitory concentration (MIC) and minimum bactericidal concentration (MBC). The antibacterial activity of the essential oil against E. coli was enhanced considerably when it was converted into a nanoemulsion, which was attributed to easier access of the essential oils to the bacterial cells. The mechanism of antibacterial activity was investigated by measuring potassium, protein, and nucleic acid leakage from the cells, and electron microscopy. Evaluation of the kinetics of microbial deactivation showed that the nanoemulsion killed all the bacteria in about 5min, whereas only a 1-log reduction was observed for pure essential oil. The nanoemulsion appeared to amplify the antibacterial activity of essential oils against E. coli by increasing their ability to disrupt cell membrane integrity.

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