Abstract: Tephritidae fruit flies usually harbour different bacterial symbionts in their digestive system. These symbionts have been known to play significant role in different fitness parameters of flies. In the present study bacterial symbionts from the mid-gut of laboratory host reared Oriental fruit fly (Bactrocera dorsalis) (Hendel) were isolated and identified using conventional culture based techniques. The mid-gut of ten days old adult female was dissected and processed aseptically and initially total viable bacterial count was determined using nutrient agar medium. Different and discrete colonies on this media were isolated and subcultured. Pure culture of each isolate was subjected to a series of different morphological, cultural and biochemical tests for identification. Total viable bacterial count was revealed as 4.5x105 CFU mL-1 in the mid-gut of B. dorsalis. Eleven bacterial genera with thirteen species were identified. The identified bacterial gerera of B. dorsalis were Listeria, Citrobacter, Moraxella, Proteus, Streptobacillus, Enterobacter, Serratia, Vibrio, Aeromonas, Klebsiella, Morganella. Experimental result reported the presence of Moraxella and Streptobacillus, Aeromonas and Morganella for the first time in mid-gut of laboratory host reared B. dorsalis.