Streptococcus pneumoniae has been rarely considered as an infectious agent in appendicitis. We report a case of a 47-year-old woman with acute appendicitis caused both by serotype 35B S. pneumoniae and Klebsiella pneumoniae. The pathway of the appendix colonisation remains unclear. It could be explain by direct infection via mucosal translocation or by hematogenous spread. Pneumococcal appendicitis could progress to perforation more frequently. The use of intraoperative samples for management of appendicitis is controversial. But, culture with appropriate media is the only mean to isolate bacteria not very often encountered in appendicitis and to identify species of epidemiologic interest as serotype 35B S. pneumoniae, a non vaccinal serotype resistant to penicillin which is considered as a potential emergent pathogen. In the case of S. pneumoniae appendicitis, it could be recommended to take complementary directed samples to understand its pathophysiology.