Longitudinal assessment of mycotoxin co-exposures in exclusively breastfed infants

Early-life development of infants may be critically affected by man-made or natural contaminants including mycotoxins. However, data on the occurrence of food contaminants in breast milk is scarce and prohibits a comprehensive exposure and risk assessment for mothers and their infants. Here, we present a longitudinal exposure assessment over the first 211 days of a single newborn girl (studyA) by measuring multiple mycotoxins in milk. Eighty-seven consecutive breast milk samples were obtained from the newborn's mother living in Austria and following a regular mixed diet. Mycotoxins were analyzed by utilizing a highly sensitive LC-MS/MS approach covering 29mycotoxins and key metabolites. In addition to this longitudinal study, three mothers provided breast milk samples each on five consecutive days, for a preliminary comparison of inter-day and inter-individual variation in exposures (studyB). StudyA revealed that mycotoxin occurrence in breast milk was limited to the emerging mycotoxins alternariol monomethyl ether (AME), beauvericin (BEA), enniatins (A, A₁, B, B₁) and to ochratoxin A (OTA), which is regulated in commercial infant food. These mycotoxins were, if present, mostly detected at very low concentrations (<10 ng/L), except AME which exceeded this concentration on two distinct days by a factor of 3x and 5x. Overall, longitudinal results indicated chronic low-dose exposure to the detected mycotoxins. Other regulated mycotoxins including the carcinogenic aflatoxins or the estrogenic zearalenone and their biotransformation products were absent in all tested samples. StudyB confirmed the results of studyA, with minimal inter-day and inter-individual variation. In addition, a preliminary correlation of OTA levels occurring in breast milk and matched urine samples was found (r = 0.64, p = 0.034) in study B. Based on the data set obtained in studyA, exposure of the infant was estimated. Exposure estimates of individual mycotoxins were on average below 1 ng/kg body weight per day. Our preliminary findings suggest that recommended maximum daily intake levels might not be exceeded in the Austrian population. However, exposure is likely to be higher in populations with lower food safety standards. In the light of co-occurrence of several emerging mycotoxins in breast milk, future studies should address low-dose mixture effects. This also includes other environmental contaminants which may be present in this bio-fluid and should involve an exposome-scale risk assessment. All these efforts must be intended to minimize exposure of mothers and infants in a window of high susceptibility.