Phytochemical Analysis, Antibacterial and Antibiofilm Activities of <i>Aloe vera</i> Aqueous Extract against Selected Resistant Gram-Negative Bacteria Involved in Urinary Tract Infections

In bacterial infections, including urinary tract infections (UTIs), the gap between the development of new antimicrobials and antimicrobial resistance is dramatically increasing, especially in Gram-negative (Gram–) bacteria. All healthy products that can be used per se or that may be sources of antibacterial compounds should be considered in the fight against this major public health threat. In the present study, the phytochemical composition of Aloe vera extract was investigated by HPLC–MS/MS, and we further evaluated its antibacterial and antibiofilm formation activity against selected resistant Gram– bacteria involved in UTIs, namely, Achromobacter xylosoxidans 4892, Citrobacter freundii 426, Escherichia coli 1449, Klebsiella oxytoca 3003, Moraxella catarrhalis 4222, Morganella morganii 1543, Pseudomonas aeruginosa 3057, and a reference strain E. coli ATCC 25922. Inhibition zones (IZs) of the extract were determined using the well diffusion method, minimum inhibitory (MIC), and bactericidal (MBC) concentration by the two-fold serial microdilution assay, and antibiofilm formation activity by the crystal violet attachment assay. Aloe-emodin and its derivatives were the major constituent (75.74%) of A. vera extract, the most important of them being aloesin (30.22%), aloe-emodin-diglucoside (12.58%), and 2′-p-methoxycoumaroylaloeresin B (9.64%). The minerals found in the extract were sulfur (S), silicon (Si), chlorine (Cl), potassium (K), and bromine (Br). Except for the clinical strain E. coli 1449, which was totally non-susceptible, A. vera demonstrated noteworthy antibacterial activity with MIC and MBC values ranging from 0.625 to 5 mg/mL and 5 to 10 mg/mL, respectively. A. vera also demonstrated dose-dependent antibacterial effects, and the reference strain E. coli ATCC 25922 was the most susceptible with MIC = 0.625 and IZ = 19 mm at 20 mg/mL. The antibiofilm formation potential of A. vera extract was strong at 2MIC and MIC (93–100% of biofilm formation inhibition), moderate at MIC/2 (32–41%), weak at MIC/4 (14–21%), and nil at MIC/8

Optimization of Ethanolic Extraction of <i>Enantia chloranta</i> Bark, Phytochemical Composition, Green Synthesis of Silver Nanoparticles, and Antimicrobial Activity

In this study, using the Box–Behnken model, we optimized the ethanolic extraction of phytochemicals from Enantia chloranta bark for the first time, assessed the composition with HPLC-MS/MS, performed the green synthesis of silver nanoparticles (AgNPs) and characterized them with UV-Vis spectrophotometry, photon cross-correlation spectroscopy, energy-dispersive X-ray fluorescence spectrometry, and Fourier transform infrared spectroscopy. The antibacterial and antibiotic-resistance reversal properties of optimized extract (O-ECB) and AgNPs were assessed on various microorganisms (15 Gram−, 7 Gram+, and 2 fungi) using the well diffusion method and microbroth dilution assay. The mechanism of action was investigated on growth kinetic and proton pumps of Escherichia coli. The in vivo antimicrobial activity and toxicity were assessed on Galleria mellonella larvae. The optimal mass yield (14.3%) related to the highest antibacterial activity (31 mm vs. S. aureus ATCC 6538) was obtained with the following operating conditions: % EtOH—100%; ratio m/v—20 g/mL; and extraction time—6 h. All the compounds identified in O-ECB were alkaloids and the major constituents were palmatine (51.63%), columbamine +7,8-dihydro-8-hydroxypalmatine (19.21%), jatrorrhizine (11.02%), and pseudocolumbamine (6.33%). Among the minerals found in O-ECB (S, Si, Cl, K, Ca, Mn, Fe, Zn, and Br), Br, Fe, and Cl were the most abundant with mean fluorescence intensities of 4.6529, 3.485,4, and 2.5942 cps/uA, respectively. The synthesized AgNPs revealed a strong absorption plasmon band between 430 and 450 nm and an average hydrodynamic diameter ×50 of 59.74 nm, and the presence of Ag was confirmed by a characteristic peak in the spectrum at the silver Kα line of 22.105 keV. Both O-ECB and AgNPs displayed noteworthy and broad-spectrum antimicrobial activities against 20/24 and 24/24 studied microorganisms, respectively, with recorded minimal inhibitory concentrations (MICs) ranging from 8 to ≥1024 µg/mL and 2 to 64 µg/mL. O-ECB and AgNPs showed antibiofilm properties and significantly enhanced the efficacy of conventional antibiotics against selected multidrug-resistant bacteria, and the mechanistic investigations revealed their interference with bacterial growth kinetic and the inhibition of H+-ATPase proton pumps. LD50s were 40 mg/mL and 0.6 mg/mL for O-ECB and AgNPs, respectively. In conclusion, the current study provides a strong experimental baseline to consider Enantia chlorantha bark and their green synthetized AgNPs as potent antimicrobial compounds in this era of antimicrobial resistance