In this study, the combinations with Olea europaea (olive) oil of fatty acids of Punica granatum, Calendula officinalis and Helianthus annuus seeds to detect interactions on Escherichia coli, Enterococcus faecalis, Staphylococcus aureus, Bacillus subtilis, Candida albicans and Candida parapsilosis were determined by antimicrobial checkerboard method. It was revealed that linoleic, oleic, palmitic and stearic acid were to be main component of O. europaea, C. officinalis and H. Annuus. By contrast, P. granatum seed oil contained more than 70% conjugated linolenic acids (CLnA), followed by linoleic acid, oleic acid, stearic and palmitic by using GC-FID. Exposed microorganisms to combinations with O. europaea oil of fatty acids of P. granatum, C. officinalis, H. annuus overnight, according to FICI (Fractional Inhibitory Concentration Index) interpretation model, remarkable IndE was observed as 71.43% against E.coli and 76.19% against S. aureus, B.subtilis, C.albicans while lower AntE noted as 28.57% against E.coli and 23.81% against S.aureus, B.subtilis, C.albicans.