This study assessed the phytochemical content and antifungal effectiveness of Ocimum basilicum seed extracts utilizing phytochemical screening, Gas Chromatography-Mass Spectrometry (GC-MS), and Fourier-Transform Infrared (FTIR) spectroscopy. Extracts were used to inhibit the germination of Aspergillus flavus and Aspergillus parasiticus spores and to limit aflatoxin production both in vitro and in stored cattle feed. Phytochemical analysis confirmed the presence of many bioactive compounds. GC-MS analysis indicated solvent-dependent variations: alcohol extract was abundant in esters and fatty acids (e.g., tridecanoic acid methyl ester 9.82%, hexadecanoic acid ethyl ester 19.86%, tridecanoic acid 29.70%), chloroform extract exhibited high levels of estragole and aromatic esters, whereas n-hexane extract contain hydrocarbons and fatty acids. FTIR spectra validated the presence of functional groups. In antifungal experiments the alcohol extract was the most efficacious, limiting spore germination of A. parasiticus and A. flavus by 80% and 85% at a concentration of 250 mg mL-1, and reducing aflatoxin formation by 86%. In untreated controls, A. flavus generated 72 µg L-1 of AFB2 and 90 µg L-1 of AFB1, whereas A. parasiticus produced 55-100 µg L-1 of AFG2, AFG1, AFB2, and AFB1. Aflatoxin levels in cattle feed controls reached approximately 100 ppb AFB2 and 110 ppb AFB1 after six months, while seed powder treatment inhibited accumulation in concentration-dependent manner; at 20% supplementation, A. parasiticus was entirely suppressed, and A. flavus aflatoxins were diminished by up to 98%. The findings illustrate the efficacy of O. basilicum seed extracts and powder as environmentally sustainable inhibitors of aflatoxin contamination in animal feed.
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