Rickettsia felis is an obligate intracellular bacterium belonging to the spotted fever group, suspected to cause a murine typhus-like illness in humans, with a cosmopolitan distribution. This study was designed to estimate presence and occurrence of this pathogen in fleas collected from dogs and cats in different areas of Italy. Two species of fleas were identified, Ctenocephalides felis (80.3%) and Ctenocephalides canis (19.7%). Overall, 320 fleas (257 C. felis and 63 C. canis) collected from 117 animals (73 dogs and 44 cats) were tested. Thirty-eight (11.9%) C. felis fleas, 13 from cats (17.6%) and 25 from dogs (10.2%) were positive for R. felis. No C. canis was positive. Fleas from cats showed a tendency to be more positive than fleas from dogs. Prevalence of R. felis among areas and within provinces of the same area was extremely variable, ranging from 0 to 35.3%. Overall, prevalence in north-eastern Italy (23.2%) was significantly higher than in south-western Italy (7.1%). This study confirmed the occurrence of R. felis in cat and dog fleas (C. felis) from Italy, similar to other European countries. The results also suggest that R. felis should be considered in the human differential diagnosis of any spotted-like fever in Italy, especially if the patient is known to have been exposed to flea bites.