Controlling vegetative cells and spores growth of Bacillus spp. using ethanolic Ketapang (Terminalia catappa L.) leaf extract

Terminalia catappa L. is a large, spreading type of tree which usually grows in tropical environment, especially at coastal area with sandy stones. The current study evaluated anti-Bacillus potential of the ethanolic ketapang (Terminalia catappa L.) leaf extract (EKLE) as antibacterial and sporicidal agent against vegetative cells and spores of Bacillus spp. The antibacterial activity of EKLE against Bacillus spp. (B. cereus ATCC33019, B. pumilus ATCC14884, B. subtilis ATCC6633 and B. megaterium ATCC14581) vegetative cells were determined by performing well diffusion assay (WDA), minimum inhibition concentration (MIC), minimum bacterial concentration (MBC) and time-kill curve analyses. The sporicidal activity was tested at different concentrations of EKLE. Then, the extract's stability in terms of antibacterial and sporicidal activities upon exposure to different temperatures and pHs were carried out. Results demonstrated inhibition zones of EKLE against Bacillus spp. was in the range of 9.25 ± 0.75 mm - 11.67 ± 0.47 mm. All vegetative cells of Bacillus spp. were inhibited with MIC values at 0.63-1.25 mg/mL and can be completely killed with MBC values of 0.63 - >5.00 mg/mL. Time-kill analysis showed all the Bacillus spp. tested can be completely killed at concentrations of 2.50-5.00 mg/mL from 1 to 4 h. EKLE concentration of 1% (w/v) completely killed all Bacillus spp. spores at different exposure time. The antibacterial and sporicidal activities of EKLE were not affected by exposure to different temperatures (4, 30, 50, 80 and 121 °C) and pHs (3, 7 and 10), revealing the stability of the extract against different conditions. In conclusion, Terminalia catappa L. leaf exhibits antibacterial and sporicidal activities against Bacillus spp., therefore, the extract can be developed as anti-Bacillus agent, paving the way for its utilization in food industry as a natural food preservative.