The present work is a trial to compare effect of short and long term administrations of sucrose as a natural widely used sweetener, aspartame as a synthetic non-caloric sweetener and stevia as a novel natural non-caloric sweetener on rats' brain hippocampus. This was achieved using 64 male albino rats. Rats were divided into four groups. The first group served as control group receiving distilled water. The second group represented the sucrose group receiving 10% sucrose solution (weight /volume). The third group served as aspartame group receiving aspartame (75 mg/kg body weight /day).The fourth group represented the stevia group and received (40 mg/kg body weight/day).Animals received different experimental treatments by oral gavage and they were dissected after a short duration of 30 days and long experimental duration of 90 days. Histological investigation of hippocampal sections of hippocampus revealed neuronal degeneration with pyknotic nuclei, dilatation and congestion in blood vessels and spongiform changes in the neuropil after both sucrose and aspartame treatments. On the other hand, stevia treatment to rats showed near to normal pattern of the granular and the neuropil. Histochemical, stained sections of Bromophenol blue stain to hippocampal tissue recorded significant decrease in protein content in hippocampus in sucrose group and aspartame groups respectively after 90 days compared with stevia and control rats. Staining hippocampus tissue of rats treated with sucrose or aspartame with Congo red stain revealed cellular homogenous pink deposits of amyloid throughout the hippocampus tissue. This was decreased and observed as limited or few amyloid depositions in stevia group compared to controls. In conclusion, Stevia, a non-calorific sweetener, is a better alternative to the synthetic sweetener aspartame because stevia has the potential to assist individuals in regulating their weights without any recognized side effects on hippocampus tissue besides its health benefits as a natural antioxidant.