In this study a total number of 15 moribund gold fish (Carassius auratus) were collected from the fish farms of Hooghly district, West Bengal, India. The bacterial infection in the eyes of the affected fishes was recorded. Isolation and biochemical characterization of the bacterial isolate was done in laboratory. The bacterial isolate (GF1) was found to be positive for catalase, methyl red, nitrate reduction test, citrate, starch hydrolysis test and negative for urease, voges proskauer, indole, oxidase, gelatin hydrolysis, casein hydrolysis and lipase test. GF1 utilized glucose and sucrose as carbon source. The bacteria could tolerate upto 10% NaCl in the nutrient broth medium. The bacteria were found to be sensitive to the recommended doses of ciprofloxacin, gentamycin, levofloxacin, streptomycin, tetracycline and nalidixic acid. Poly acrylamide gel electrophoresis (PAGE) analysis revealed that the isolate contained 11 discrete bands ranging from 24.853 to 73.941kDa proteins. Phylogenetic analysis of the 16S rRNA gene sequence was also done. Based on the morphological, biochemical and phylogenetic analysis the GF1 isolate was identified as Bacillus sp.