(S91F and D95G in GyrA and S87R in ParC). Both isolates were genotyped using N. gonorrhoeae multi-antigen sequence typing and the analysis showed that the ST225 which represented an emerging widespread multi-resistant clone that has also been associated with reduced susceptibility to ceftriaxone. We recommend continued surveillance of this pathogen to assess the efficacy of anti-gonococcal antibiotics in Brazil.Key words: Neisseria gonorrhoeae -fluroquinolone resistance -gyrA and parC -genotyping -MAST Two male patients, a 24-year-old and a 32-year-old, were treated at the sexually transmitted diseases (STD) clinic after presenting with urethral discharge and dysuria. The patients provided urethral culture swabs for Neisseria gonorrhoeae and were treated with a single oral dose of ciprofloxacin (500 mg). Because the symptoms persisted, a single dose of ceftriaxone (250 mg) intramuscular was administrated, which cured all of the symptoms. Tests later confirmed that both culture swabs were positive for N. gonorrhoeae. Both patients were human immunodeficiency virus-negative, although one of them was venereal disease research laboratory-reactive (1/4). One patient reported sexual contact with 10 casual partners in the last three months before infection, whereas the other patient reported two casual partners. It was not possible to know if patients had sexual contact in another state of Brazil or country.The gonococci were screened using several tests, including beta-lactamase BD BBL TM (BD, Franklin Lakes, NJ, USA), Serovar (Phadebact ® GC Serovar Test, Bactus AB, Huddinge, Sweden) and E-test ® (AB Biodisk, Solna, Sweden). The E-test ® was performed using a media containing a GC Medium Base (BD-Difco TM ) and 1% defined growth supplements to ciprofloxacin, ofloxacin, chloramphenicol, ceftriaxone, azithromycin, penicillin and tetracycline. Reference values for sensitivity, reduced sensitivity or resistance were set according to the Clinical and Laboratory Standards Institute + Corresponding author: wianfe@yahoo.com.br WAF, CMF and FGN contributed equally to this work. Received 24 September 2010 Accepted 6 June 2011 (CLSI 2005a) and (Van Dick et al. 1999), specifically to chloramphenicol. The ATCC 49226 strain was used as a control for antibiotic quality.The strains were not beta-lactamase producers and were classified in the WII-III serogroup with Bo, Bp, Br, Bs and Bt serotypes. The isolates showed resistance to ciprofloxacin, ofloxacin (> 32.00 µg/mL) and chloramphenicol (3 µg/mL and 2 µg/mL). One strain was classified as having chromosomally-mediated resistance to penicillin [non-PPNG, non-TRNG with penicillin, minimal inhibitory concentrations (MIC) of ≥ 2 µg/mL and tetracycline MIC of ≤ 2 µg/mL] and reduced susceptibility to tetracycline (0.75 µg/mL and 0.500 µg/mL). Both strains were susceptible to azithromycin (0.125 µg/ mL and 0.190 µg/mL). MIC values were 0.064 µg/mL and 0.032 µg/mL. The agar dilution method was used to confirm MICs of ciprofloxacin and ofloxacin at > 32.00 µg/mL. The major difference (p...