The purpose of this study was to determine the levels of trans-octadecenoic acid (C18:1-trans) and trans-isomers of linoleic acid (18:2-trans), as well as long-chain polyunsaturated fatty acids (LC-PUFA), in the plasma from infants of adolescent mothers at 3 months of age, exclusively breastfed, and the relationship with the levels of the same isomers in plasma and milk of the mothers. Samples of blood and mature milk were obtained from 49 healthy adolescent mothers and their exclusively breastfed infants treated at the Instituto Fernandes Figueira-Fundação Oswaldo Cruz (IFF-FIOCRUZ) in Rio de Janeiro, Brazil. trans-Fatty acids (TFA) were analyzed by gas chromatography. The results of this study showed low levels of TFA in milk (1.53%), maternal plasma (0.50%), and plasma of infants (0.74%). The results show that, although TFA have been found in the plasma of the studied infants, the LC-PUFA levels are kept within normal limits. No association between TFA presence and parameters of nutritional status of the infants was observed, probably due to the low levels of these fatty acids found in this study.