BACKGROUND Ciprofloxacin replaced chloramphenicol (C), the best choice of antibiotic in the treatment of enteric fever, when C-resistant enteric fever emerged and caused outbreaks in different parts of the world. C-sensitive S. enterica serovar Typhi emerged again due to withdrawal of the antibiotic pressure. OBJECTIVE To assess the in vitro efficacy of C against Salmonella enterica serovar Typhi isolates (1991-2003). METHODS A total of 464 blood culture isolates of S. enterica serovar Typhi were subjected to C susceptibility by disc diffusion and agar dilution methods using Mueller-Hinton agar. The antibiotic susceptibility of S. enterica serovar Typhi isolates obtained in the year 2002 and 2003 was determined using ampicillin, cotrimoxazole, ciprofloxacin, nalidixic acid, ceftriaxone and cefotaxime, in addition to C. Escherichia coli strain ATCC 25922 was used as the control. Changes in C sensitivity of the isolates were analyzed using chi2 test with Yates correction. CONCLUSIONS All the isolates of 1991 were C-resistant with minimum inhibitory concentration values (MICs) of 2000-5000 mg/ml. In the following years decrease in frequency of C resistance was noticed: 1992 (50%), 1993 (32%), 1994 (27%) and 1995 (05%). The isolates of 1996-99 and 2001 were 100% C-sensitive. In 2000, sensitivity was also high (79%). The strains isolated in the year 2002 and 2003, showing reduced susceptibility of ciprofloxacin, were nalidixic acid resistant, but sensitive to the third-generation cephalosporins (ceftriaxone and cefotaxime). The MICs for C-sensitive isolates (1991-2003) ranged 0.1-5 mg/ml. Results suggest the necessity for re-evaluation of C therapy in typhoid fever.