Early-life is a critical window in offspring development. An unhealthy diet during this period can negatively impact offspring metabolic function and eating behaviour later in life​. Microbiota-targeted interventions hold promise for the treatment of aberrant functioning of eating behaviour. This study evaluates the impact of an early-life high-fat/high-sugar (HFHS) diet on eating behaviour later in life, and whether this can be attenuated by microbiota-targeted strategies.Male and female C57BL/6J mice were exposed to either control or HFHS diet from birth for 5 weeks. Afterwards, all mice received control diet. Two groups with early-life HFHS diet were supplemented with a prebiotic (Fructo-Oligosaccharides+Galacto-Oligosaccharides, FOS+GOS) or a bacteria (B. longum APC1472) throughout the study. Meal pattern, saccharin and food preference were investigated at 6 and 11 weeks.Five-week-old female and male pups exposed to early-life HFHS showed greater weight-gain compared to control pups with no microbiota-targeted intervention effects. Body weight was normalised after 1 and 2 weeks respectively, after switching to control diet. Early-life HFHS diet increased meal size, eating rate and total HFHS intake in 11-week-old females. Early-life HFHS increased preference towards palatable food at 6 and 11 weeks in both sexes whilst simultaneously increasing food grinding behaviour. APC1472 alleviated HFHS-induced alterations in food preference and food grinding behaviour in both sexes. APC1472 and FOS+GOS alleviated the alterations in meal pattern in females.These findings demonstrate that an unhealthy diet in early-life has enduring effects on eating behaviour later in life in a sex-specific manner, which can be alleviated by microbiota-targeted strategies.