Meloidogyne incognita (root-knot nematode) is a devastating soil-borne pathogen which can infect almost all cultivated plants around the globe, expediting huge pecuniary losses. The purpose of current study was to use the aqueous root extract of Glycyrrhiza glabra for synthesizing silver nanoparticles (GRAgNPs) and assess their nematicidal potential against M. incognita by in vitro methods, including hatching inhibition and mortality assays. The active uptake of FITC labeled GRAgNPs by the nematode and their effect on the expression of selected genes involved in oxidative stress and DNA damage repair were also studied. An HRTEM micrograph confirmed their spherical morphology with sizes ranging from 9.61 nm to 34.735 nm. Complete inhibition of egg-hatching was observed after 48 h of treatment with as low as 10.0 ppm of GRAgNPs. In addition, 100% mortality was recorded at the lowest dose of 6.0 ppm, after 12 h of treatment. The LC-50 for GRAgNPs was found to be 0.805 ± 0.177 ppm at p < 0.0001, R² = 0.9930, and α = 0.05. The expression of targeted genes (skn-1, mev-1, sod-3, dhs-23, cyp-450, xpa, cpr-1, gst-n, and ugt) was significantly enhanced (1.09–2.79 folds), at 1.0 ppm (α = 0.05, 95% CI) GRAgNPs treatment. In conclusion, GRAgNPs performed efficaciously and considerably in contrast to chemical nematicide and commercial silver nanoparticles (CAgNPs) and might be used as a promising alternative as relatively lower concentration and short exposure time were enough to cause higher mortality and nanotoxicity in nematodes.

Bio-fabrication of green or plant extract-based silver nanoparticles has garnered much praise over the past decade as the methodology is environment-friendly, undemanding, non-pathogenic, and economical. In the current study, leaves of Eurale ferox (Makhana), considered as waste, were used for the bio-fabrication of silver nanoparticles (ELAgNPs). Various analytical techniques including UV–VIS spectroscopy, Field emission scanning electron microscopy equipped with an energy dispersive X-ray spectrometer (FESEM-EDX), Particle size analyzer (PSA), Fourier transform infra-red spectroscopy (FTIR) and high-resolution transmission electron microscopy (HRTEM) were used for their characterization. Their antibacterial efficacy was examined against gram positive bacterium, Bacillus subtilis and gram negative bacterium, Escherichia coli. The antioxidant potential of the ELAgNPs was compassed by 2, 2 diphenyl-1-picryl hydrazyl (DPPH; λ_max = 517 nm) assay, H₂O₂ (λ_max = 230 nm) and OH⁻ (λ_max = 520 nm)-based radical scavenging assays. The cytotoxicity was checked against the VERO cell line using 3-[4, 5-dimethyl thiazol-2-yl]-2, 5 diphenyl tetrazolium bromide (MTT) assay. A mean particle size of 26.51 ± 8.87 nm with a size distribution of 7.08–53.94 nm was obtained using HRTEM. The ELAgNPs exhibited dose-dependent antibacterial efficacy with a maximum zone of inhibition (ZOI) of 21.98 ± 0.59 mm against B. subtilis and of 16.46 ± 0.22 mm against E. coli at 500 ppm after 24 h of incubation. The median lethal concentration for the cytotoxicity analysis was found to be 9.54 ± 0.35 ppm, 120.9 ± 6.31 ppm, and 20.74 ± 0.63 ppm for ELAgNPs, commercial silver nanoparticles (CAgNPs), and silver nitrate (SN), respectively. The ordinary one-way ANOVA results exhibited a significant decrease in cell viability after 72 h of incubation at p < 0.05, α = 0.05. In conclusion, the ELAgNPs showed good antibacterial, radical scavenging and dose-dependent cytotoxicity against the VERO cells. Therefore, these could be used for biomedical applications. Phyto-constituents present in the plant not only act as reducing agents but also as stabilizing and coating agents, and the availability of a wide range of metabolites makes the green approach more promising.

The existing treatments against Trypanosoma evansi are faced with several drawbacks, such as limited drug options, resistance, the relapse of infection, toxicity, etc., which emphasizes the necessity for new alternatives. We synthesized novel metal-based antiparasitic compounds using chitosan, hydroxychloroquine (HC), and ZnO nanoparticles (NPs) and characterized them for size, morphology, chemical interactions, etc. Molecular docking and protein interaction studies were performed in silico to investigate the inhibitory effects of HC, zinc-ligated hydroxychloroquine (HCZnONPs), and chitosan-zinc-ligated hydroxychloroquine (CsHCZnONPs) for two key proteins, i.e., heat shock protein 90 (Hsp90) and trypanothione reductase associated with T. evansi. In vitro trypanocidal activity and the uptake of zinc ions by T. evansi parasites were observed. The formulation was successfully synthesized, as indicated by its size, stability, morphology, elemental analysis, and functional groups. CsHCZnO nanoparticles strongly inhibit both Hsp90 and trypanothione reductase proteins. The inhibition of Hsp90 by these nanoparticles is even stronger than that of trypanothione reductase when compared to HC and HCZnONPs. This suggests that the presence of polymer chitosan enhances the nanoparticles’ effectiveness against the parasite. For the first time, CsHCZnO nanoparticles exhibited trypanocidal activity against T. evansi, with complete growth inhibition being observed at various concentrations after 72 h of treatment. Fluorescent microscopy using FluoZin-3 on T. evansi culture confirmed the presence of zinc on the surface of parasites. This innovative approach has shown promising results in the quest to develop improved antiparasitic compounds against T. evansi with enhanced effectiveness and safety, highlighting their potential as therapeutic agents against trypanosomiasis.

Isometamidium chloride (ISM) is a trypanocide for the prophylactic and therapeutic use against vector-borne animal trypanosomosis (mainly Surra caused by Trypanosoma evansi) and African animal trypanosomosis caused by T. congolense/T. vivax/T. brucei). ISM was found to be an efficient trypanocide for therapeutic/prophylactic use against trypanosomosis; however, it produces some local and systemic detrimental effects in animals. We synthesized isometamidium chloride-loaded alginate gum acacia nanoformulation (ISM SANPS) to lessen the detrimental side effects of isometamidium chloride (ISM) while treating trypanosomal diseases. We intended to determine the cytocompatibility/toxicity, and DNA deterioration/chromosomal structural or number changes (genotoxicity) of ISM SANPs using mammalian cells in a concentration-dependent manner. Apurinic/apyrimidinic (AP) sites are one of the major types of DNA lesions formed during base excision and repair of oxidized, deaminated, or alkylated bases. The intensity of the cellular AP site is an excellent marker of the deterioration of DNA quality. We thought it pertinent to quantify the AP sites in ISM SANPs-treated cells. Our investigations established a dose-dependent cyto-compatibility or toxicity and DNA impairment (genotoxicity) in ISM SANPs-treated horse peripheral blood mononuclear cells. ISM SANPs were biocompatible at various concentrations tested on the mammalian cells.