Integrated Experimental and Computational Evaluation of the Gut‐Targeted Anticoccidial Potential of <i>Agaricus bisporus</i> Methanolic Extract

Abstract

Coccidiosis caused by Eimeria spp. is a major constraint to global poultry production, causing economic losses exceeding USD 13 billion annually. Growing drug resistance from prolonged anticoccidial use highlights the need for safer, eco-friendly alternatives. Edible mushrooms, such as Agaricus bisporus, are rich in bioactive phytochemicals, yet their anticoccidial potential remains underexplored. This study investigated the methanolic extract of A. bisporus (ABE) for inhibitory effects on Eimeria papillata oocyst sporulation and interactions with essential apicomplexan enzymes. ABE was chemically characterized by GC-MS, identifying 25 compounds, predominantly lipids, terpenoids, and steroids, with major constituents including 9,12-octadecadienoic acid (Z,Z)- (24.65%), vitamin E (17.91%), and cis-vaccenic acid (15.84%). In vitro assays demonstrated concentration-dependent inhibition of oocyst sporulation, achieving 83.44% at 72 h and 87.97% at 96 h (200 mg/mL), surpassing amprolium and comparable to disinfectants. Mechanistic insights via deep-learning-rescored molecular docking of major compounds against enoyl-ACP reductase (ENR) and lactate dehydrogenase (LDH) suggested multiligand interactions, whereas ADMET predictions indicated favorable gut retention and safety profiles. These findings position ABE as a promising, eco-friendly anticoccidial candidate. Future in vivo studies are warranted to validate its efficacy and practical application in poultry production.