This study aimed to determine the effect of maternal exposure to a glyphosate-based herbicide, during pregnancy and lactation, upon body parameters, glucose homeostasis and liver histopathology in adult offspring male mice that were fed on a high fat diet. C57bl/6 female mice received 0.5% glyphosate (Roundup Original DIâ) in their drinking water (GBH group) or filtered water (CTRL group), during pregnancy and lactation. From postnatal day 60 (PND60) until PND150, offspring (F1) received a high-fat diet (GBH-HFD-F1 and CTRL-HFD-F1 groups). At PND 150, GBH-HFD-F1 mice presented lower body weight gain and retroperitoneal and perigonadal fat weights than mice in the CTRL-HFD-F1 group. Although both groups presented similar values for glycemia and insulinemia, the GBH-HFD-F1 group had a higher tolerance to glucose and higher insulin sensitivity, when compared to the CTRL-HFD-F1 group. Both groups demonstrated hepatic steatosis, although this condition was milder in the GBH-HFD-F1 animals. Therefore, mice exposed to glyphosate during gestation and lactation and fed on a high-fat diet in adulthood, exhibited alterations in glycemic and lipid metabolism, we speculate that these metabolic changes may be associated with decreased body weight and fat accumulation in these mice, Further studies are needed to elucidate the underlying mechanisms responsible for these effects.