This study was conducted in 78 roosters Hisex white layer to investigate the possible effect of use or misuse of antibiotics on liver and kidneys function as well as some other important organs. Three antibiotics of varying groups i.e., tylosin, ciprofloxacin and oxytetracycline were chosen for the study.  Antibiotics were daily orally administered in different doses as described in the experimental body. The possible adverse effect of antibiotics on liver and kidneys functions were followed by testing liver and kidneys serum biomarkers such as Glutamate oxido transaminase (GOT), Glutamate pyruvate transaminase (GPT), acid phosphatase (ACPase), alkaline phosphatase (ALKPase), total protein, albumin, globulin etc. Antibiotics residues clearance in plasma samples were also studied and correlated to administered dose. Histopathological investigations were further supported our biochemical data on the adverse effect of antibiotics on kidney, liver, heart, and other tissues. The present study clearly concluded a direct adverse effect of antibiotics, regardless of the chosen group, on liver and kidney and heart function.