2 Salmonella Typhimurium

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1 Salmonella Typhimurium 2 1) 1) 2) 1) 2) Salmonella Typhimurium (ciprofloxacin (CPFX) MIC 16 mg/ml) S. Typhimurium 2 fosfomycin (FOM) 1 PCR gyra parc RAPD-PCR DNA S. Typhimurium Key words: Salmonella Typhimurium, gyra parc Salmonella 1) Salmonella Typhimurium 2), 3) S. Typhimurium definitive phage type 104 (DT104) 4) Salmonella 5) 8) ( ) TEL: FAX: ogura@tamanan-hp.com S. Typhimurium 9) 2 S. Typhimurium DNA I kg, (Fig. 1): (WBC) 9,100/ ml, CRP 9.5 mg/dl 36 Vol. 16 No

2 97 Fig. 1. Clinical course of case 1 and case 2. (Fig. 1): fosfomycin (FOM) 3 g/ / S. Typhimurium Vol. 16 No

3 kg, 40.3 (Fig. 1): 7,300/ml, CRP 8.5 mg/dl (Fig. 1): 40 FOM 600 mg/ / 3 FOM 1,500 mg/ / S. Typhimurium II. 1. 1, 2 Salmonella sp Micro Scan Walk/Away NC5J DADE 3. (MIC) ampicillin (ABPC), cefotaxime (CTX), kanamycin (KM), streptomycin (SM), minocycline (MINO), chloramphenicol (CP), FOM, norfloxacin (NFLX), ofloxacin (OFLX), levofloxacin (LVFX), sparfloxacin (SPFX), tosufloxacin (TFLX), ciprofloxacin (CPFX) 13 CLSI (Clinical and Laboratory Standards Institute) 10) 4. DNA DNA Kado and Liu 11) sodium lauryl sulfate (SDS) DNA (0.9 ) 5. Randomly amplified polymorphic DNA-PCR (RAPD-PCR) DNA DNA Okazaki 12) PCR DNA (2.4 ) 6. Giraud 13) DNA gyra, gyrb, parc, pare (quinolone resistance-determining regions, QRDR) DNA PCR PCR DNA 14) III. 1, 2 (Table 1) Salmonella sp. S. Typhimurium (O-4: i: 1, 2) (Table 2): 1, 2 MIC (mg/ml) NFLX (32, 64), OFLX (8, 8), LVFX (8, 8), SPFX (16, 32), TFLX (16, 16), CPFX (16, 16), ABPC ( 128, 128), SM 128, 128), MINO (64, 32), CP ( 128, 128) Table 1. Di#erentiation of fluoroquinoloneresistant strains of Salmonella Typhimurium strains Characteristics Reaction Strain 1 Strain 2 Oxidase ONPG IPA VP Indole Lysin decarboxylase Ornithine decarboxylase Arginine decarboxylase Urease Malonate utilization Glucose Adonitol Arabinose Inositol Rhamnose Sorbitol Sucrose 38 Vol. 16 No

4 99 Table 2. Strain MICs of 13 antimicrobial agents against fluoroquinolone-resistant strains of Salmonella Typhimurium MIC ( mg/ml of a NFLX OFLX LVFX SPFX TFLX CPFX ABPC CTX KM SM MINO FOM CP S. Typhimurum S. Typhimurum a Abbreviations: NFLX, norfloxacin; OFLX, ofloxacin; LVFX, levofloxacin; SPFX, sparfloxacin; TFLX, tosufloxacin; CPFX, ciprofloxacin; ABPC, ampicillin; CTX, cefotaxime; KM, kanamycin; SM, streptomycin; MINO, minocycline; FOM, fosfomycin; CP, chloramphenicol. ABPC, SM, MINO, CP RAPD-PCR (Fig. 2): 1, 2 DNA RAPD-PCR DNA QRDR 1, 2 DNA gyra 2 parc 1 GyrA ParC 80 (Table 3) Fig. 2. Plasmid profiles (A) and RAPD profiles (B) of fluoroquinolone-resistant strains of S. Typhimurium. Lane 1, S. Typhimurium strain 1; lane 2, S. Typhimurium strain 2. IV. ABPC, CP, SM, Salmonella Salmonella FOM 1 Table 3. Strain Mutations in quinolone resistance-determining regions of gyra (GyrA) and perc (PerC) of fluoroquinolone resistant Salmonella Typhimurium strains gyra/(gyra) parc/(parc) S. Typhimurium a GGC GAT TCC GCA GTG TAT GAC ACC GGC GAC AGC GCC TGC (NA s ) (Gly) (Asp) (Ser) (Ala) (Val) (Tyr) (Asp) (Thr) (Gly) (Asp) (Ser) (Ala) (Cys) S. Typhimurium T A C 1 (Phe) (Asn) (Arg) S. Typhimurium T A C 2 (Phe) (Asn) (Arg) a Nalidixic acid-susceptible strain. Vol. 16 No

5 100 9) 2 Salmonella 2 S. Typhimurium DNA RAPD DNA (Fig. 2) 2 ABPC, CP, SM, NFLX Salmonella O-4 S. Typhimurium S. Typhimurium DNA 15) gyra, gyrb, parc, pare 1, 2 DNA gyra 2 parc 1 3 IV B gyrb pere S. Typhimurium gyra perc S. Typhimurium 1) : ) Threlfall, E. J., J. A. Frost, L. R. Ward, et al Increasing spectrum of resistance in multiresistant Salmonella Typhimurium. Lancet 347: ) Glynn, M. K., C. Bopp, W. Dewitt, et al Emergence of multidrug-resisutant Salmonella enterica serotype Typhimurium DT 104 infections in the United States. N. Eng. J. Med. 338: ) Wall, P. G., E. J. Threllfall, L. R. Ward, et al Multiresistant Salmonella Typhimurium DT104 in cats: A public health risk. Lancet 348: ) : ) Salmonella serovar Typhimurium definitive type : ) : ) : ) Salmonella enterica serotype Typhimurium definitive phage type : ) Clinical and Laboratory Standards Institute Performance standards for antimicrobial susceptibility testing: 15th informational supplement CLSI/NCCLS M100-S15. Clinical and Laboratory Standards Institute, Wayne, Pa, USA. 11) Kado, C. I., S. T. Liu Rapid procedure for detection and isolation of large and small plasmids. J. Bacteriol. 145: ) Okazaki, M., T. Watanabe, K. Morita, et al Molecular epidemiological investigation 40 Vol. 16 No

6 101 using a randomly amplified polymorphic DNA assay of Burkholderia cepacia isolates from nosocomial outbreaks. J. Clin. Microbiol. 37: ) Giraud, E., A. Brisabois, J.-L. Martel, et al Comparative studies of mutations in animal isolates and experimental in vivo- and in vivoselected mutants of Salmonella spp. suggest a counterselection of highly fluoroquinoloneresistant strains in the field. Antimicrob. Agents Chemother. 43: ) Lee, L., C. R. Connell, S. L. Woo, et al DNA sequencing with dye-labeled terminators and T7 DNA polymerase: E#ect of dyes and dntps on incorporation of dye-terminators and probability analysis of termination fragments. Nucleic Acids Res. 20: ) : Two Cases of Infantile Diarrhea Caused by Fluoroquinolone-resistant Salmonella Typhimurium Ken-ichi Ogura, 1) Fuyuhiko Saito, 1) Koji Morita 2) 1) Department of Clinical Laboratory, Tama-Nambu Chiiki Hospital 2) Department of Microbiology, Kyorin University Scool of Health Sciences Two infants, 7-year-old female and 7-month-old male, were hospitalized because of diarrhea. Stool cultures of both patients were positive for Salmonella Typhimurium with high-level fluoroquinoloneresistance (ciprofloxacin MIC of 16 mg/ml). They were treated with fosfomycin, and recovered completely. Two S. Typhimurium strains, from above two cases of infantile diarrhea, had some identical features including di#erentiation of S. Typhimurium strains resistance pattern, plasmid profile, and genotype on randomly amplified polymorphic DNA-PCR. Furthermore, mutations in quinolone resistance-determining regions of GyrA (amino acids change at Ser-83 and Asp-87) and ParC (amino acid change at Ser-80) were also coserved in two S. Typhimurium strains. These results suggest that two infantile diarrhea patients investigated in this case study were infected with the identical clone of fluoroquinolone-resistant S. Typhimurium. Though a conclusive evidence of infection routes were unknown, the pet-related infections were suspected in both cases of infantile diarrhea. Extensive surveillance for fluoroquinolone-resistance Salmonella serovars is necessary. Vol. 16 No