This research was carried out to study for the first time the colonization, the biology and to collect first baseline data on susceptibility status of Phlebotomus papatasi the vector of Cutaneous Leishmaniasis (CL) in Sudan. Samples of P. papatasi females were collected from Surogia village (located at the north site of Khartoum State, Sr), White Nile forest (White Nile State, WN) and Dinder National Park located at southern east of Sudan (Gedaref State, DNP). The P. papatasi females were collected with different methods using both light and sticky oil traps. A colony of P. papatasi was established using standard protocols for rearing sandflies with minor modifications. The investigations on P. papatasi resistance to four insecticides(DDT, permethrin, malathion and propoxur) were carried out using WHO standard susceptibility test. For more investigation also a biochemical analysis was conducted to investigate the activities of four enzyme systems involved in insect metabolism and able the insect to acquire resistance to insecticide (acetylcholinesterase (AChE), non-specific esterases (EST), glutathione-S-transferases (GST) and cytochrome p450 monooxygenases (Oxy)).