Genotyping of <i>Echinococcus granulosus</i> and Evaluation of <i>Moringa oleifera</i> Leaf Extract as a Novel Protoscolicidal Agent

Cystic echinococcosis is a globally prevalent zoonotic infection, including in the Kurdistan region of Iraq. It arises when Echinococcus granulosus eggs are ingested, leading to the larval stages that mainly effect the internal organs of humans and livestocks. This study aimed to explore the genetic variability and sequence polymorphisms of E. granulosus and to evaluate the protoscolicidal potential of Moringa oleifera leaf extract. Protoscoleces PSCs were isolated from hydatid cysts collected from infected humans, sheep, cattle and goat. Among the 73 hydatid cysts examined, 63 (86.3%) were fertile. DNA was extracted from PSCs, the mitochondrial cytochrome oxidase subunit 1 (cox1) gene was amplified for molecular characterization. DNA sequencing and phylogenetic analyses were performed to identify E. granulosus genotypes. Phylogenetic analysis demonstrated the predominance of G1 and G3 genotypes. In the experimental assays, PSCs were treated with ethanolic M. oleifera leaf extract. GC–MS profiling revealed major bioactive constituents (phytol, eugenol acetate, trans-isoeugenol and vitamin E). The addition treatment included four concentrations of the extract were 250, 400, 500, and 750 mg/mL across various exposure periods 5, 15 and 30 minutes, and demonstrated strong protoscolicidal activity, achieving 100% mortality at 750 mg/mL after 30 min., while the lower mortality rate found at the concentration 250mg/ml was 13%. Scanning electron microscopy PSCs were demonstrated ultrastructural alterations with including loss of hooks, shedding of microtriches, cellular shrinkage, and lysis it was reflecting severe structural damage. Overall, these results suggest that M. oleifera leaf extract represents a promising alternative protoscolicidal agent.