It is not well known if Burkholderia cepacia can be found in fish or water. This is because bacteria may enter into the water through sewage, animal waste, and soil runoff. Therefore, the current study was conducted to assay the incidence of biochemically identified Burkholderia cepacia in seventy-five Nile Tilapia and twenty-five water samples collected from Elmanzala Lake, Egypt, with API 20 NE and its antibiotic resistance. Three B.cepacia isolates were identified from 100 samples with a prevalence of 3%. Two from fish (only gills) with a prevalence of 2.7%(2/75) and one from the water sample with a prevalence of 4%(1/25). Only one isolate was MDR with a 0.43 MAR index. Besides, all isolates were susceptible to levofloxacin, meropenem, and chloramphenicol, while, recovered isolates were resistant to ceftazidime (100%), penicillin G(66.6%), and trimethoprim/ sulfamethoxazole (33.4%). This study concluded that MDR B.cepacia is important in public health and is considered a human health issue. Meropenem, chloramphenicol, and levofloxacin are used as drugs of choice for the treatment of B.cepacia.