In Vitro and In Silico Anticoccidial Activity of Opuntia ficus-indica Seed Oil and Hexane Extract: Comprehensive GC-MS Profiling and Mechanistic Insights

The anticoccidial potential of Opuntia ficus-indica seed oil (OFI-SO) and its hexane extract from seed press cake (OFI-HexPC) was evaluated against Eimeria spp . using integrated in vitro and in silico approaches.Sporulated Eimeria oocysts were exposed to increasing concentrations (2–64 mg/mL) of OFI-SO and OFI-HexPC in Hank’s balanced salt solution. Oocyst viability was determined by microscopic counting, while membrane integrity was assessed by quantifying the release of 273 nm-absorbing substances. Toltrazuril (25 mg/mL) was used as a positive control.GC-MS profiling of both extracts revealed linoleic acid (54.2%) and oleic acid (25.7%) as the main bioactive fatty acids.GC–MS profiling revealed that linoleic acid (69.63%) was the major fatty acid in OFI-SO, while oleic acid (44.20%) predominated in OFI-HexPC. Both OFI-SO and OFI-HexPC significantly reduced oocyst counts and induced a dose-dependent increase in DNA release.In silico molecular docking was performed against three key Eimeria enzymes (EtDHODH, CDPK, and PKA). Identified fatty acids showed moderate binding affinities through hydrophobic interactions. Redocking validation (RMSD < 2 Å) confirmed the reliability of the docking protocol. ADMET predictions indicated favorable pharmacokinetics and safety profiles.These findings suggest that OFI-SO and OFI-HexPC act via a dual mechanism involving membrane disruption and enzymatic inhibition, supporting their potential as natural alternatives to synthetic anticoccidials in poultry production.