Aflatoxins comprise a group of mycotoxins that are found in the environment. Exposure to aflatoxins has been reported to cause serious health problems in humans. Since aflatoxin M-1 (AFM(1)) is secreted in breast milk, the exposure of infants to this toxin is an important concern. The aim of this study was to determine the prevalence, levels of, and factors associated with the presence of AFM(1) in breast milk of mothers in Fethiye, Turkey. Breast milk samples were taken from 100 mothers who had given birth over the period of October-November 2017. The AFM(1) content of the samples was determined via enzyme-linked immunosorbent assay. The lowest limit for milk samples in the Ridascreen (R) AFM(1) commercial test kit is 5 ng/L. Because of this, AFM(1) levels below 5 ng/L in the breast milk samples were assessed as negative. Of the breast milk samples tested, 53 were positive. The average AFM(1) amount in the positive samples was 6.36 ng/L (ppt; range 5.10-8.31 ng/L). Mothers who were housewives, lived in damp, humid houses, or ate spices or dried fruits and vegetables had significantly greater prevalence of AFM(1) in their breast milk than those who were employed, did not report dampness or mold in the home, or did not eat spices or dried fruits and vegetables. AFM(1) in breast milk could be an important risk factor for infant health. Informing the public about food safety could reduce the amount of AFM(1) being transferred into breast milk via food channels.