Background: Fish are considered as bio-indicators of the health of aquatic ecosystems. Recently, native fish species were declining due to anthropogenic pollution and records of fish availability in a specific area of the Padma Riverine System aided in understanding the current status of fish and which fish need conservation. Methods: The fish diversity in Murshidabad district’s Padma River was studied for twelve months, from October 2021 to September 2022, with the help of local fishermen. Freshly caught fish is immediately transported to a laboratory, identification is followed by Talwar and Jhingran (1991) and Rahman (2005) techniques. Result: A total of 49 fish species belonging to 11 orders, 24 families were recorded. Siluriformes (17 species) had the most dominance, followed by cypriniformes (12 species) and anabantiformes. Among the species collected, eight were threatened, thirty were less threatened, one was extremely rare and one had not yet had their status determined. These fishes were classified as threatened (16.33%), less threatened (61.23%) and exceedingly rare (2%). Based on the findings, it is concluded that the Padma River could be used as a refuge for the conservation of Murshidabad’s threatened freshwater fishes.