The study of the conditions which affect the pharmacological activity of medicinal plants is the basis for the development of phytomedicine. This study aimed at identifying some optimal conditions for antibacterial activity of extracts of Harungana madagascariensis. The leaves, bark, and roots were harvested in Douala, Littoral Region of Cameroon, in August 2019 at three times of the day: early in the morning (6 a.m.), at midday (12 p.m.), and in the afternoon (6 p.m.). They were dried at room temperature and ground to give a fine powder. Each powder was macerated in methanol for 72 h and boiled in distilled water and palm wine. The obtained extracts were evaluated for their antibacterial activity by broth microdilution method on 31 clinical and 4 reference bacterial strains. The results show that the best extraction yield was obtained using the bark extracted from palm wine by decoction. The methanol bark extract was found to be more active than other solvents (8 and 512 µg/mL). In fact, the activity of the 6 a.m. extract was significant on 4 of the 35 strains tested (MIC< 100 µg/mL) and moderate on 27 of the 35 strains tested (100≤ MIC≤ 625 μg/mL). The activity of the noon extract was significant on 8 strains and moderate on 23 strains and, finally, the activity of the extract of 6 p.m. was significant on 9 strains and moderate on 25 strains. The extract harvested at 6 p.m. inhibited the growth of the 35 strains tested and revealed significant activity (MIC <100 μg/mL) on Staphylococcus aureus, Escherichia coli, Citrobacter freundii, Serratia marcescens, Salmonella enterica serovar typhi, and Yersinia enterocolitica, making this hour the best harvesting time. This study further confirms that the bark of H. madagascariensis is a potent antibacterial agent, being better still when harvested at 6 p.m. and extracted with methanol.