Bacterial leaf blight (BLB), caused by Xanthomonas oryzae pv. oryzae (Xoo), poses a serious threat to rice cultivation. This study presents the green synthesis of zinc oxide nanoparticles (ZnO NPs) using an aqueous leaf extract of the medicinal plant Centella asiatica (L.) Urban and evaluates their potential as dual-function nanopesticides. The synthesized CA-ZnO NPs exhibited high crystallinity, a hexagonal to quasi-spherical morphology, and nanoscale dimensions (~22.5 nm), as confirmed by UV–Vis spectroscopy, XRD, FTIR, SEM, TEM, and SAED analyses. These nanoparticles demonstrated potent antibacterial activity against a highly virulent, field-derived Thai Xoo strain, with a minimum inhibitory concentration (MIC) of 8 µg/mL. Mechanistic investigations revealed substantial membrane disruption, intracellular nanoparticle penetration, and elevated reactive oxygen species (ROS) generation in treated cells. Cytotoxicity testing using human dermal fibroblasts (HDFs) revealed excellent biocompatibility, with no statistically significant reduction in cell viability at concentrations up to 500 µg/mL. In contrast, viability markedly declined at 1000 µg/mL. These findings underscore the selective antibacterial efficacy and minimal mammalian cytotoxicity of CA-ZnO NPs. Overall, CA-ZnO NPs offer a promising green nanopesticide platform that integrates potent antibacterial activity with biocompatibility, supporting future applications in sustainable crop protection and biomedical nanotechnology.

Chitosan nanoparticles (Ch NPs) are versatile nanomaterials with expanding agricultural and biomedical applications, highlighting the need for reproducible, low-cost, and scalable synthesis methods to ensure their safe and widespread use in biological systems. This study presents a simple ionic-gelation protocol using a serological pipette–needle dropwise system that minimizes reagent waste and requires no sophisticated equipment. The synthesized Ch NPs were characterized by UV–Vis spectroscopy, ESEM, TEM, EDS, DLS, XRD, and FTIR, confirming nanoscale size, strong positive surface charge, and characteristic chitosan–TPP interactions. To establish a standardized biological safety assessment framework, three representative bioassays were implemented across microbial, plant, and mammalian systems. Antibacterial testing against Xanthomonas oryzae pv. oryzae (Xoo) using a resazurin-based microdilution assay revealed a minimum inhibitory concentration (MIC) of 128 µg/mL, whereas bulk chitosan showed no inhibition up to 512 µg/mL. Phytotoxicity and seed germination assays on rice (Oryza ‘KDML105’) demonstrated no inhibitory effects on germination, with over 90% germination by day 3 and significantly enhanced seedling growth parameters (p < 0.05) at 64–128 µg/mL, indicating non-phytotoxicity. MTT assays confirmed that Ch NPs were non-toxic to both human skin cell lines (HDF and HaCaT) across 2.5–160 µg/mL, showing enhanced cell viability in HDF cells at specific concentrations and stable viability in HaCaT cells, indicating overall biocompatibility. Importantly, all bioassays were conducted under aligned concentration ranges to enable cross-system comparison and reproducibility. This integrated workflow links nanoparticle synthesis with a standardized, multi-system evaluation strategy, supporting the safe application of Ch NPs in biological systems.

Bacterial leaf blight (BLB), a destructive disease of rice caused by Xanthomonas oryzae pv. oryzae (Xoo), continues to limit rice productivity worldwide. Although biologically synthesized zinc oxide nanoparticles (ZnO NPs) have been extensively investigated, knowledge regarding the antibacterial activity and biocompatibility of commercially available ZnO NPs is still limited. In this study, commercial ZnO NPs were systematically characterized and evaluated for their antibacterial mechanisms and biocompatibility in mammalian cells. FE-SEM and TEM analyses revealed irregular polyhedral, hexagonal, and short rod-like morphologies with an average particle size of ~33 nm, consistent with crystallite sizes estimated by XRD. The nanoparticles exhibited pronounced antibacterial activity against Xoo, with a minimum inhibitory concentration (MIC) of 16 µg/mL and a clear dose-dependent response. Mechanistic assays confirmed multifaceted bactericidal actions involving membrane disruption, ROS generation, Zn²⁺ release, and ultrastructural damage. Biocompatibility testing in human dermal fibroblasts showed enhanced proliferation at 8–32 µg/mL, no cytotoxicity up to 256 µg/mL, and reduced viability only at ≥512 µg/mL. These findings represent the first mechanistic evaluation of commercial ZnO NPs against Xoo, together with cytotoxicity assessment in mammalian cells, highlighting their structural distinctness and dual functionality that combine potent antibacterial activity with minimal mammalian cytotoxicity. Overall, the results underscore their potential as safe nanobiocontrol agents for sustainable rice disease management.