Ozioma Forstinus Nwabor

Background: Avian pathogenic Escherichia coli (APEC) is a leading cause of colibacillosis in poultry. Piper betle L. is a medicinal plant rich in bioactive compounds including hydroxychavicol that possess potent antibacterial activity. This study aimed to investigate the efficacy of a P. betle L. leaf nanoemulsion (NEPE) and hydroxychavicol against multidrug-resistant APEC isolates. Methods: In vitro and in silico analysis of NEPE and hydroxychavicol against APEC were determined. Results: The nanoemulsion exhibited potent antibacterial activity, with MIC and MBC values of 0.06–0.25% v/v and 0.125–0.25% v/v, respectively. The MIC and MBC values of hydroxychavicol against isolates ranged from 0.25 to 1.0 mg/mL. A time–kill assays revealed rapid bactericidal effects of both compounds, achieving a ≥3-log reduction within 4 h at 4 × MIC. Scanning electron microscopy demonstrated that APEC cells treated with hydroxychavicol exhibited filamentous cells with incomplete septa. Molecular docking and dynamics simulations of hydroxychavicol against APEC cell division proteins were investigated. According to the binding energy, hydroxychavicol exhibited the highest affinity with ZapE, FtsW, FtsX, FtsZ, and FtsA, respectively. However, the FtsA protein showed the least protein conformational change throughout the 5000 ns simulation, reflecting a highly stable conformation. Conclusions: These confirm the potential stability of protein and ligand, as supported by molecular dynamics simulation. The results suggested the potential of NEPE and hydroxychavicol, which may have promising antibacterial potential that can be used to inhibit APEC growth.

Background and Aim: Staphylococcus aureus is a zoonotic pathogen with significant public health and economic implications. Its ability to tolerate environmental stress and regulate virulence through quorum sensing contributes to its persistence and pathogenicity. Rhodomyrtus tomentosa and its bioactive compound rhodomyrtone have demonstrated antimicrobial properties against Gram-positive, multidrug-resistant bacteria. This study aimed to evaluate the effects of these agents on quorum sensing inhibition and stress tolerance in S. aureus and Pseudomonas aeruginosa, providing insights into their potential as alternative antimicrobial strategies. Materials and Methods: The anti-quorum sensing activity of R. tomentosa extracts was assessed using Chromobacterium violaceum as a bioindicator. In addition, the effects on P. aeruginosa swarming motility were evaluated. Stress tolerance in S. aureus was examined by subjecting treated cells to acidic (pH = 5.0), alkaline (pH = 9.0), osmotic (7.5% NaCl), heat (43°C), and oxidative (1 mM H₂O₂) stress conditions. The survival rates were determined through colony-forming unit (CFU) counts following treatment with rhodomyrtone and ethanol leaf extracts. Results: The ethyl acetate fraction of R. tomentosa leaf extract exhibited the highest violacein inhibition, followed by the ethanol extract. At 256 μg/mL, these extracts permitted P. aeruginosa colony formation but inhibited its swarming motility. Regarding stress tolerance, no surviving S. aureus cells were detected under any stress condition after 3–6 h of treatment with 2 × minimum inhibitory concentration (MIC) (1 μg/mL) of rhodomyrtone. In addition, 4 × MIC (128 μg/mL) of the ethanol leaf extract inhibited pathogen survival under all tested stress conditions except for alkaline and oxidative stresses. Conclusion: The findings suggest that R. tomentosa extracts and rhodomyrtone effectively inhibit quorum sensing and stress tolerance, offering a promising alternative antimicrobial approach. These compounds could be utilized in veterinary medicine and food safety to mitigate zoonotic pathogen contamination and combat antibiotic-resistant infections. Keywords: anti-quorum sensing, Pseudomonas aeruginosa, rhodomyrtone, Rhodomyrtus tomentosa, Staphylococcus aureus, stress tolerance.

Aim: Polymeric scaffolds were developed fortified with nanovesicle-encapsulated individual curcumin (CUR) and tetrahydrocurcumin (THC) for improved therapeutic efficacy due to their low stability and efficacy in native form. Method: Nanovesicle-encapsulated individual CUR and THC were fabricated using thin-film hydration techniques and characterized. Results & conclusion: CUR/THC in native and vesicle-encapsulated form demonstrated diminished LPS-instigate nitric oxide (NO) levels in macrophage cells in a concentration-dependent demeanor. However, vesicle-encapsulated CUR/THC inhibited NO production at lower concentrations, compared with the native CUR/THC form. Furthermore, the scaffold fortified with vesicle-encapsulated CUR/THC demonstrated improved physical properties with excellent antioxidant, biocompatibility, and human keratinocyte cell proliferation ability. The results recommended that nanovesicle-encapsulated THC can be retained as a potential substitute for CUR with improved therapeutic efficacy.

The emergence and spread of polymyxin resistance, especially among Klebsiella pneumoniae isolates, threaten the effective management of infections. This study profiled for polymyxin resistance mechanisms and investigated the activity of polymyxins plus vancomycin against carbapenem- and polymyxin-resistant K. pneumoniae . The entire genome sequences of seven isolates were profiled for resistance and virulence determinants. The effects of combination therapy were evaluated using the checkerboard technique, time-kill assay, and population profile analysis. Protein profiles of the isolates treated with monotherapy were compared to that of combination therapy. The whole-genome sequencing data revealed that the isolates harbored β-lactams, carbapenems, aminoglycoside, fluoroquinolones, macrolides, and tetracycline resistance genes, with several virulence-associated genes. The plasmids including the bla _OXA-232 -bearing ColKP3 plasmid were also identified in our isolates. Profiling for polymyxin resistance mechanism revealed a missense mutation in the crrB gene that resulted in a Q180L variant that conferred a deleterious effect on protein function. The combination assay indicated fractional inhibitory concentration index ranging from 0.31 to 1.13, whereas the time-kill assay demonstrated synergistic log reduction in colony-forming units per milliliter. Furthermore, population analysis profiling using dual antibiotics indicated enhancement in bacterial log reduction at lower antibiotics concentrations, compared to higher concentrations of single polymyxins. For protein profiling, 796 proteins were identified, and 56 and 94 of them were increased and decreased in the combined drug treatment groups, respectively, while other differentially produced proteins were detected in all treatment groups, except for the control group. The results demonstrated that the vancomycin combination might benefit the antimicrobial activities of polymyxins. IMPORTANCE: This study provides insights into the mechanisms of polymyxin resistance in K. pneumoniae clinical isolates and demonstrates potential strategies of polymyxin and vancomycin combinations for combating this resistance. We also identified possible mechanisms that might be associated with the treatment of these combinations against carbapenem- and polymyxin-resistant K. pneumoniae clinical isolates. The findings have significant implications for the development of alternative therapies and the effective management of infections caused by these pathogens.

Quercus infectoria (Qi), a traditional herbal plant with broad-spectrum of activities on multidrug-resistant bacteria has been develop as hand sanitizer applications. Antimicrobial activity was evaluated using agar-well diffusion and broth microdilution method. Bactericidal activity was determined following the European Standard 1276 antibacterial suspension test. Neutralization assay was performed to assess anti-respiratory syncytial virus. Safety, stability, and skin permeation of Qi hand gel was investigated. Qi hand sanitizer gel inhibited microorganisms ranging from 99.9–99.999% against Enterococcus faecalis, Staphylococcus aureus, methicillin-resistant S. aureus, S. epidermidis, S. pseudintermedius, S. saprophyticus, Streptococcus pyogenes, Acinetobacter baumannii, Escherichia coli, Klebsiella pneumoniae, Pseudomonas aeruginosa, and Candida albicans. A significant reduction in main human dermatophytes including Microsporum canis, Microsporum gypseum, and Talaromyces marneffei of approximately 50% was observed (pQi hand sanitizer gel inactivated more than 99% viral particles entering human laryngeal epidermoid carcinoma cells in a dose-dependent manner. Scanning electron micrographs further illustrated that Qi hand sanitizer gel disrupted microbial cell membrane after 1-min contact time resulting in cell death. Qi hand sanitizer gel delivered emollient compounds through simulated human skin layers and showed no cytotoxicity on fibroblast cells. Moreover, Qi hand sanitizer gel demonstrated stability under extreme conditions. Qi hand sanitizer gel was able to inhibit various microorganisms including bacteria, dermatophytes, and virus.

Biofilm-mediated infections are critical to public health and a leading cause of resistance among pathogens, amounting to a prolonged hospital stay and increased mortality rate in the intensive care unit. In this study, the antibacterial and antibiofilm activities of rifampicin or carbapenem monotherapies were compared with rifampicin and carbapenem combination therapies against rifampicin-resistant and carbapenem-resistant Acinetobacter baumannii isolates. Among 29 CRAB isolates, 24/29 (83%) were resistant to rifampicin, with MIC values between 2–256 µg/mL. Checkerboard assays disclosed that combination therapies at FICIs between 1/8 and 1/4 improved the activity of carbapenems at subinhibitory concentrations. Time-kill kinetics indicated a 2- to 4-log reduction at 1/2 MIC rifampicin + 1/4 MIC carbapenem and 1/4 MIC rifampicin + 1/4 MIC carbapenem against the isolates, with the MIC values ranging from 2–8 µg/mL. The MTT assay revealed a dose-dependent decrease of the cell viability of established bacterial biofilm at 4 MIC rifampicin + 2 MIC carbapenems, with a percentage reduction of 44–75%, compared with monotherapies at 16 MIC. Scanning electron microscopy further confirmed bacterial cell membrane disruption, suggesting a synergism between carbapenem and rifampicin against a representative isolate. The findings demonstrated that the combination of rifampicin with carbapenems could improve antibacterial activities and eradicate established Acinetobacter baumannii biofilm.

Series of works on the biological properties of rhodomyrtone, a major active component of Rhodomyrtus tomentosa leaf has been reported. In this study, rhodomyrtone-rich microwave-assisted extract from R. tomentosa leaf (RT-MAE) before formulation was profiled for phytochemical composition using gas chromatography-mass spectrometry (GC-MS) and targeted for anti-inflammatory activity with topical application. The formulation was assessed for bioactivity, biocompatibility, and safety on human volunteers. The GC-MS spectra indicated a rich content of phytosterols, sesquiterpenes, and acylphloroglucinols. At concentrations 50–600 ng/mL, RT-MAE before formulation treated macrophages showed >86% cell viability. Nitric oxide production significantly decreased by 19.26–88.36% following the treatment with 50–600 ng/mL RT-MAE before formulation (p < 0.001). Moreover, a significant (p < 0.01) concentration-dependent downregulation of lipopolysaccharide-induced IL-6, IL-1β, iNOS, and COX-2 mRNA was observed in RT-MAE before formulation treated macrophages. The extract demonstrated minimum inhibitory (MIC) and minimum bactericidal concentration (MBC) at 0.25–4 and 1–16 μg/mL, respectively. The antioxidant activity of RT-MAE before formulation indicated IC₅₀ of 0.16 and 0.06 ug/mL for DPPH and ABTS assays, respectively. Ex-vivo diffusion of rhodomyrtone from formulation showed steady-state flux 64.935 ng/cm² h, with >80% viability on HaCaT cells. Safety assessment of formulation containing RT-MAE after formulation was conducted on healthy volunteers. Hematological and biochemical analysis showed no significant difference between test and baseline (p < 0.05). The results suggested that the formulation can be effectively employed as a topical inflammatory agent.

Biofilms associated with multidrug-resistant (MDR) Acinetobacter baumannii on medical devices remain a big clinical problem. Antibiotic susceptibility tests were performed with eight commonly employed antibiotics against clinical isolates. The effects of antibiotics in combination with well-characterized lytic phage T1245 were studied to assess their antibacterial and anti-biofilm efficacy. Ceftazidime, colistin, imipenem, and meropenem significantly reduced bacterial density up to approximately 80% when combined with phage T1245, compared with control. Phage T1245 in combination with ceftazidime, colistin, and meropenem at subinhibitory concentrations demonstrated significant reduction in biomass and bacterial viability of 3-day established biofilms, compared with antibiotic alone. In addition, electron microscopy further confirmed the disruption of biofilm structure and cell morphology upon treatment with phage T1245 and antibiotics, including ceftazidime, colistin, and meropenem. Combined treatment of phage T1245 with these antibiotics could be employed for the management of A. baumannii infections and eradication of the bacterial biofilms.

Ensuring the safety of food products is critical to food production and processing. In food processing and production, several standard guidelines are implemented to achieve acceptable food quality and safety. This notwithstanding, due to human limitations, processed foods are often contaminated either with microorganisms, microbial byproducts, or chemical agents, resulting in the compromise of product quality with far-reaching consequences including foodborne diseases, food intoxication, and food recall. Transitioning from manual food processing to automation-aided food processing (smart food processing) which is guided by artificial intelligence will guarantee the safety and quality of food. However, this will require huge investments in terms of resources, technologies, and expertise. This study reviews the potential of artificial intelligence in food processing. In addition, it presents the technologies and methods with potential applications in implementing automated technology-aided processing. A conceptual design for an automated food processing line comprised of various operational layers and processes targeted at enhancing the microbial safety and quality assurance of liquid foods such as milk and beverages is elaborated.