The farming of Rainbow trout, an exotic fish has advanced on a huge scale in north western regions of India which provides the best suitable climate and topography for its culture. The advancement in farming is challenged by the infectious diseases. The isolation of Stenotrophomonas maltophilia, an opportunistic pathogen has been reported from the cultured channel catfish in china, and cultured African catfish in West Bengal, India. The present study reports isolation of Stenotrophomonas maltophilia from the kidney of Rainbow trout, Oncorhynchus mykiss along with its biochemical, 16 s rRNA sequencing and phylogenetic analysis. Antibiotic susceptibility of isolates to 10 antimicrobial drugs was investigated using Disc diffusion method. The isolates were found to be resistant to multiple drugs, characterised by resistance to all Beta-lactams (Methicillin and Ampicillin), Macrolides (Erythromycin, Azithromycin and Rifampicin), aminoglycosides (Streptomycin, Amikicin) except Gentamicin and susceptible to Tetracycline and Chloramphenicol antibiotics. The auto-aggregation capability of the isolates showed 55.02 ± 5.9 and 49.52 ± 7.6% auto-aggregation after 24 h of incubation, respectively. The isolates showed 32.77 ± 6.78 and 29.41 ± 7.71 % hydrophobicity after 2 h of extraction with xylene, respectively. The 16 s rRNA sequences of the were submitted to the GenBank database with the accession numbers OP647420 and PP077305.