The study was carried out to investigate the antibiotic sensitivities of bacteria isolated and identified from Chrysichtys nigrodigitatus and their antibiotic susceptibility pattern determined. A total of 45 samples of skin, gill and intestine of Chrysichtys nigrodigitatus were obtained from Ogun River, Akomoje in Abeokuta, Ogun State and analyzed microbiologically. The mean values of characteristics measured for the body weight, total length, and standard length for the 15 fish samples are 147.99±66.41, 26.55±5.55 and 20.05±4.13 respectively while the physical and chemical parameters of Ogun River, Akomoje during the study is presented as follows; Water Temperature (29.0°C), pH (7.8), Dissolved Oxygen (4.9mg/l) and Ammonia (2.3ug/dL). The total mean bacteria count in the gill samples was 1.06 ± 0.20X106CFU/g, while in the intestine samples is 1.36 ± 0.27X106CFU/g and in the skin samples it is 0.77 ± 0.21X106CFU/g. All the samples collected showed bacterial growth on Eosin methylene Blue (EMB) and Salmonella-Shigella Agar. The 45 samples gave 135 isolates; 28 (15.3%) Escherichia coli, 30 (22.2%) Staphylococcus
aureus, 34 (25.2%) Bacillus sp, 30 (22.2%) Salmonella sp, 13 (9.6%) Pseudomonas auregenosa. Five isolates were selected characterized by molecular method using 16S rRNA gene sequencing. The antibiotic test of these isolates revealed their sensitivity to ciprofloxacin, pelfloxacin, and tarivid. It was indicated that all the bacterial isolate was susceptible to Ciprofloxacin and Sparfloxacin. Escherichia coli were also susceptible to Pefloxacin, Ciprofloxacin and Tarivid. E. coli was also resistance to Streotomycin, Augmentin and Amoxacillin, Gentamycin, Septrin and Chloranphenicol.