Didelphis albiventris
is a marsupial with synanthropic habits, whose contact with domestic animals and humans is increasing. They are considered potential disseminators of diseases, present as definitive hosts or reservoirs of various pathogenic agents. The present study aim to report, for the first time, the parasitism by
Ctenocephalides felis
in
D. albiventris
in Rio Grande, Southern Rio Grande do Sul state, Brazil. An adult female White-eared Opossum (
D. albiventris
), killed by being run over, was collected on the roads of the city of Rio Grande, Rio Grande do Sul, Brazil and sent to the laboratory for necropsy. During external inspection, a flea was found, which was stored in 70
o
alcohol until taxonomic identification, being classified as
C. felis
. This study reports the occurrence of parasitism by
C. felis
in
D. albiventris
for the first time in the location studied, demonstrating that these arthropods circulate between the wild and urban environments, and play an important role in the cycle of zoonotic pathogens among wild fauna, companion animals and humans.