Fishes and fishery by-products are a good source of proteins, high-quality amino acids, fatty acids, and minerals. Fishes are next only to meat and poultry as a staple food of most of the countries. Aquaculture has thus become an important activity in many countries which contributes to 25% of the total world seafood supply (Srinivasan and Saranraj, 2017). The World Bank Group Agriculture action plan highlights the expected projection in global fish supply (1,86,842 MT) and consumption (1,51,771 MT) by 2030. The remaining will be wasted. The dumping of this waste into the sea is a very unscientific practice as it undergoes decomposition and generates toxic by-products. This waste can be converted into a more functional form, hydrolysate, which possesses a high nitrogen content (Wangkheirakpam et al., 2019). The present study focused on finding the potential of fish protein hydrolysate (FPH) as a low-cost alternative of peptone in microbial culture media. The efficiency of fish protein hydrolysate (Catla catla and Labeo rohita) as a peptone source for bacteria (pathogenic and non-pathogenic strains) was compared with commercial peptone. The growth of bacterial strains was found better in FPH as compared to that in commercial media. The growth of Acinetobacter baumanii A87 was obtained 17% higher in formulated media as compared to that in commercial media. Similarly, among the pathogenic strains, Salmonella bongori BR1859 exhibited 10% higher growth in formulated media. Conversion of fish visceral waste into protein hydrolysate is thus an environment-friendly approach and also produces reasonable raw material for microbial media formulation (Parvathy et al., 2018). Such an application of fish visceral waste has been stated earlier; however, a comparison of both pathogenic and non-pathogenic strains together has not been reported yet.