Tsetse flies Diptera: Glossinidae) are the biological vectors of the trypanosomes responsible sleeping sickness in humans and African animal trypanosomiasis AAT) in domestic animals in sub-Saharan Africa. Ecological factors, climate change and habitat fragmentation affect distribution in tsetse flies. but little information is available in the conserved areas. The objective of our study is to update information on the distribution and density of tsetse flies in the protected area of Folonzo, in the southwest of Burkina Faso. For the sampling and monitoring of tsetse flies, the transect method was used. Each transect consisted of 5 biconical traps set from the bank of the stream to the savannah. The total of 25 traps 5 transects) were collected every 24 hours and for 5 days per season. The overall density of the 4 tsetse species initially encountered G. tachinoides; G. palpalis gambiensis, G. morsitanssubmorsitans and G. medicorum) in 2012, was reduced by 87.11% in 2019, p <0.001) .However, no capture of G. m. submorsitans was only observed in 2019. The influence of the season effect on tsetse density has not been verified. This study shows that tsetse populations in conservation areas are also affected by the impact of climate change and human pressure. These pressures can lead to extinction G. m. submorsitans) , scarcity G. tachinoides) and the resilience G. p. gambiensis) of some species. Knowledge of tsetse distribution is an important component in understanding the dynamics of trypanosome transmission in order to effectively control AAT.