The inoculated medium was incubated in a shake incubator (The Lab Companion IS-971 (R/RF) Floor Model Incubated Shaker, GMI Inc., USA) at 30°C and agitation speed of 140 rpm for 48 h.
Then the medium was incubated at 26°C in incubator (Memmert Type INB500, Germany) and was examined at the different life stages of Leishmania tropica and was identified under 100x magnification of microscope for every 24 hours till up to 10 days.
Briefly, 1 ml of complete medium was incubated with PRP-SF bioscaffolds for 24 hours.
Each inoculated medium was incubated at agitation speed 150 rpm at 37 °C for overnight.
Then, the medium was incubated at 45 °C for 72 h.
The medium was incubated at 37 °C, 220 rpm, for 12 h.
After the inoculation, the production medium was incubated at 37 °C and pH 6.5 for 4 days.
The inoculated medium was incubated at 30 °C and 200 r/min in the dark for 5 days.
The inoculated medium was incubated at 28°C ± 2°C and agitated at 150 rpm for 10 days.
The solid LB medium was incubated at 37 °C for overnight since it is easier to get highly active bacteria in solid LB than in liquid LB.
After inoculation with 5 mLs obtained from the overnight culture of each isolate, the medium was incubated under static conditions (no shaking) at temperature 30 °C and pH 7.
