Background: Cholera, an acute intestinal infection caused by the bacterium Vibrio cholera and is a historically feared epidemic diarrheal disease that remains a major public health problem in many parts of Africa, Asia and Latin America. Vibrio cholerae 01 has two main Inaba and Ogawa serotypes and is a member of the Vibrio family. This organism is transmitted through drinking water, food and vegetable thah has been contaminated with feces and also through raw seafood or inadequately heated food such as oyster. The aim of the study was isolate Vibrio cholerae 01 (Vibrionaceae) in the Karon River.Methods: In four month (April, may, June and July 2010), a total 100 samples of water from Karoon River Ahvaz were collected. During the study period the recorded river temperature was about 25-28oC and pH ranged from 7 to 8. Water samples were collected, concentrated by centrifuge (6000 rpm), and enriched in alkaline peptone water (APW (1% peptone, 1% NaCl, pH 8.4–8.6)) for isolation of Vibrio spp. After 24h incubation, sampeles were transported onto thiosulphate citrate bile sucrose, and morphological colonies compatible with Vibrio were characterized by oxidase test and agglutinated with antiserum (Difco, Detroit, MI, USA) for serotype determination. Also V. cholerae biochemical tests with API 20E (BioMerieux, Marcyl’Etoile, France).Results: Vibrio cholerae strains were isolated from the 100 karonriver water samples. Amoung of these, 8 were found to contain; Vibrio cholerae O1 (inaba) and 7 were V. parahaemolyticus strains.Conclusion: The priorities for cholera control remain public health interventions through improved water and sanitation, improved surveillance and access to health care facilities, and further development of appropriate vaccines.