172( 38 ) THE JAPANESE JOURNAL OF ANTIBIOTICS 57 2 Apr. 2002 1 19 6 2002 5 8 4 254 254 (PBP) 90 83 65 142 PBP pbp1a, pbp2x, pbp2b 121 (49%), pbp1a, pbp2x 30 (12%), pbp2x, pbp2b 16 (6%), pbp2x 61 (24%), pbp1a 1 (1%), pbp2b 1 (1%) 24 (9%) cefditoren cefcapene panipenem, biapenem, imipenem meropenem tosufloxacin sparfloxacin mefa, ermb mefa 74 (29%) ermb 134 (53%), mefa ermb 10 (4%) 36 (14%) 55 6 17 30.9% 40 8 14.5% 9 6 10.9% 15 5 9.1%

Apr. THE JAPANESE JOURNAL OF ANTIBIOTICS 57 2 173( 39 ) Penicillin resistant Streptococcus pneumoniae (PRSP) 6 11/29 37.9% 9 3 10.3% 15 3 10.3% PRSP 29 Pulsed Filed Gel Electrophoresis 15 1) penicillin-resistant Streptococcus pneumoniae (PRSP) 1 1988 2) 3,4) 5) I. 1. 2002 5 8 4 6 254 penicillin-susceptible S. pneumoniae (PSSP) 24 penicillin-intermediate S. pneumoniae (PISP) 230 1 5 2. PCR Polymerase chain reaction (PCR) (PRSP) PCR (LytA) PCR 3,4,6) PCR PSSP, PISPS PRSP pbp1a, pbp2x, pbp2b PSSP 1 2PISP 3 PRSP 3. penicillin G (PCG), piperacillin (PIPC), amoxicillin (AMPC), cefaclor (CCL), cefixime (CFIX), cefdinir (CFDN), cefteram (CFTM), cefpodoxime (CPDX), cefditoren (CDTR), cefcapene (CFPN), imipenem (IPM), panipenem (PAPM), meropenem (MEPM), biapenem

174( 40 ) THE JAPANESE JOURNAL OF ANTIBIOTICS 57 2 Apr. (BIPM), levofloxacin (LVFX), ciprofloxacin (CPFX), tosufloxacin (TFLX), sparfloxacin (SPFX), clarithromycin (CAM), azithromycin (AZM), minocycline (MINO), clindamycin (CLDM) 22 5% 80 C MIC 37 C, 5% 2 4. 55 5. PRSP PRSP PRSP 29 Sma I 7) Pulsed Field Gel Electrophoresis (PFGE) II. 1. 1556 57 52 19 55 254 MIC2000, MIC2000, SEPTOR System, SN 2. 90 (34%), 83 (33%), 65 (26%), 14 (6%), 2 (1%)0 5 139 (55%), 6 10 15 (6%), 11 64 40 (16%), 65 53 (22%), 7 (3%) 0 5 0 25 (10%), 1 45 (18%), 2 15 (6%), 3 31 (12%), 4 19 (7%), 5 4 (2%) 60 (24%), 106 (41%), 62 (24%), 9 (4%), 17 (7%) 3. PCG PSSP 4 (21%) PISP 15 (79%), PSSP 64 (45%) PISP 77 (54 ), PSSP 23 (15%) PISP 96 (85%), PSSP 7 (14%) PISP 45 (86%), PSSP 18 (56%) PISP 14 (44%), PSSP 93 (18%) PISP 434 (82%) PSSP 24 PISP50 230 b - pbp1a, pbp2x, pbp2b Table 1 pbp1a, pbp2x, pbp2b (PRSP) 121 (48%), pbp1a, pbp2x (PISP) 30 (12%),

Apr. THE JAPANESE JOURNAL OF ANTIBIOTICS 57 2 175( 41 ) Table 1. Isolation frequency of abnormal penicillin binding protein (PBP) genes in Streptococcus pneumoniae isolated from hospitals in Gifu area. Table 2. Isolation frequency of macrolide-resistant genes in Streptococcus pneumoniae isolated from hospitals in Gifu area. pbp2x, pbp2b (PISP) 16 (6%), pbp2x (PISP) 61 (24%), pbp1a (PISP) 1 (1%), pbp2b (PISP) 1 (1%), (PSSP) 24 (9%) pbp1a, pbp2x, pbp2b 50% pbp1a, pbp2x pbp2b (p 0.05) mefa, ermb Table 2 mefa74 (29%), ermb 134 (53%), mefa, ermb 10 (4%), 36 (14%) mefa, ermb (p 0.05) 4. S. pneumoniae PSSP, PISP PRSP Table 3 MIC 50, MIC 90,

176( 42 ) THE JAPANESE JOURNAL OF ANTIBIOTICS 57 2 Apr. Table 3. Antibacterial activity of antibacterial compounds against Streptococcus pneumoniae clinical isolates of normal and harboring abnormal penicillin binding protein (PBP) genes.

Apr. THE JAPANESE JOURNAL OF ANTIBIOTICS 57 2 177( 43 ) Table 3. (Continued)

178( 44 ) THE JAPANESE JOURNAL OF ANTIBIOTICS 57 2 Apr. Table 3. (Continued)

Apr. THE JAPANESE JOURNAL OF ANTIBIOTICS 57 2 179( 45 ) Table 3. (Continued)

180( 46 ) THE JAPANESE JOURNAL OF ANTIBIOTICS 57 2 Apr. Table 3. (Continued)

Apr. THE JAPANESE JOURNAL OF ANTIBIOTICS 57 2 181( 47 ) Table 3. (Continued) MIC rangepbp1a PBP2b 1 MIC range PCG PSSP 24 1 2 PISP pbp1a; 1 pbp2x; 61 pbp2b; 1 pbp1a pbp2x; 30 pbp2x pbp2b; 16 3 PRSP 121 PCG PSSP, PISP PRSP MIC 90 0.03 m g/ml, 0.06 m g/ml 0.5 m g/ml 2 mg/ml PIPC AMPC PCG CCL CFIX PSSP 1 PISP PRSP CFDN CPDX PSSP PISP CFTM PSSP PISP CDTR PSSP, PISP PRSP MIC 90 0.03 m g/ml, 0.25 m g/ml 1 mg/ml 1 m g/mlcfpn CDTR PRSPMIC 8 mg/ml 2 4 mg/ml 3 CFPN PISP MIC 4 mg/ml 1 PRSP 16 m g/ml 5 8 mg/ml 3 PSSP, PISP PRSP PSSP MIC 90 0.015 m g/mlprsp MIC 90 0.125 m g/ml 0.5 m g/ml

182( 48 ) THE JAPANESE JOURNAL OF ANTIBIOTICS 57 2 Apr. PSSP 8 16pbp1a 2 1 LVFX CPFX MIC 90 1 mg/ml 2 mg/mltflx SPFX MIC 90 0.125 mg/ml 0.5 mg/ml CAM AZM PSSP MIC 50 0.03 mg/ml 0.125 m g/ml CLDM MINO 5. S. pneumoniae Table 4 PSSP CAM, AZM CLDM MIC 90 0.06 m g/ml, 0.125 m g/ml 0.06 mg/ml mefa CLDM PSSP CAM AZM ermb mefa ermb 6. Table 5 55 17 6 (17/55, 30.9%), 40 (8/55, 14.5%), 9 (6/55, 10.9%), 15 (5/55, 9.1%) PRSP 6 (11/29, 37.9%), 9 (3/29, 10.3%), 15 (3/29, 10.3%) 7. PRSP 29 PFGE Table 6 15 Table 4. Antibacterial activity of macrolide antibiotics against Streptococcus pneumoniae clinical isolates of normal and harboring mefa and ermb gene.

Apr. THE JAPANESE JOURNAL OF ANTIBIOTICS 57 2 183( 49 ) Table 5. Serotypes of Streptococcus pneumoniae isolates in Matsubara Otorhinolaryngology Clinic. H, M B N 5 43 III. 1977 APPELBAUM PRSP 8) 1988 2) PRSP PRSP 3,4,11) 1999 81 11) CDTR MIC 8 mg/ml 2 4 mg/ml 3 PRSP CFPN MIC 4 m g/ml 1 PISP, 16 mg/ml 5 8 m g/ml 3 PRSP PRSP CDTR 1 mg/ml

184( 50 ) THE JAPANESE JOURNAL OF ANTIBIOTICS 57 2 Apr. Table 6. Diversity of PRSP isolates in Matsubara Otorhinolaryngology Clinic using pulsed field gel electrophoresis. Time above MIC 0.5 0.7 mg/ml CDTR 30% pharmacokinetics/pharmacodynamics (PK/PD) PK/PD 19 6 23 14 9 15 PRSP 9,12) 19 23 14 40 9 151 PFGE PRSP 7) PFGE PRSP PK/PD 9) 1 1 10) 3000 0 10,598.18 km 2, 210 199 /km 2

Apr. THE JAPANESE JOURNAL OF ANTIBIOTICS 57 2 185( 51 ) 13) 1) KLUGMAN, K. P.: Pneumococcal resistance to antibiotics. Clin. Microbiol. Rev. 3: 171 196, 1990 2) b - 1 62: 682 683, 1988 3) UBUKATA, K.; Y. ASAHI, K. OKUZUMI, et al.: Incidence of penicillin-resistant Streptococcus pneumoniae in Japan, 1993 1995. J. Infect. Chemother. 2: 77 84, 1997 4) UBUKATA, K.; T. MURAKI, A. IGARASHI, et al.: Identification of penicillin and other b -lactam resistance in Streptococcus pneumoniae by PCR. J. Infect. Chemother. 3: 190 197, 1997 5) FELMINGHAM, D.; R. R. REINERT, Y. HIRAKATA & A. RODLOFF: Increasing prevalence of antimicrobial resistance among isolates of Streptococcus pneumoniae from the PROTEKT surveillance study, and compatative in vitro activity of the ketolide, telithromycin. J. Antimicrob. Chemother. 50 (Suppl. S1): 25 37, 2002 6) NAGAI, K.; Y. SHIBASAKI, K. HASEGAWA, et al.: Evaluation of the primers for PCR to screen Streptococcus pneumoniae isolates, b-lactam resistance and to detect common macrolide resistance determinants. J. Antimicrob. Chemother. 48: 915 918, 2001 7) SHIMADA, J.; N. YAMANAKA & M. HOTOMI: Household transmission of Streptococcus pneumoniae among siblings with acute otitis media. J. Clin. Microbiol. 40: 1851 1853, 2002 8) APPELBAUM, P. C.; A. BHAMJEE, J. N. SCRAGG, et al.: Streptococcus pneumoniae resistant to penicillin and chloramphenicol. Lancet 2: 995 997, 1977 9) 1998 2000 Streptococcus pneumoniae 51: 60 70, 2003 10) HORVAT, R. T.; N. E. KLUTMAN, M. K. LACY, D. GRAUER & M. WILSON: Effect of duplicate isolates of methicillinsusceptible and methicillin-resistant Staphylococcus aureus on antibiogram data. J. Clin. Microbiol. 41: 4611 4616, 2003 11) Jpn. J. Antibiotics 53: 652 659, 2000 12) Streptococcus pneumoniae 51: 551 560, 2003 13) pp. 3 6 2001

186( 52 ) THE JAPANESE JOURNAL OF ANTIBIOTICS 57 2 Apr. EPIDEMIOLOGY OF Streptococcus pneumoniae ISOLATES IN GIFU PREFECTURE Gifu Working Group of Forum on Microbial Resistance HIROSHIGE MIKAMO Department of Obstetrics and Gynecology, Division of Organ Pathobiology, Gifu University School of Medicine HIROYUKI SUEMATSU Clinical laboratories, Chuno General Hospital KAORI TANAKA and KUNITOMO WATANABE Division of Anaerobe Research, Life Science Research Center, Gifu University HARUKI SAWAMURA Gifu University School of Medicine SHIGENORI MATSUBARA Matsubara Otorhinolaryngology Clinic YOKO MATSUKAWA Clinical laboratories, Gifu Prefectural Tajimi Hospital SHIOMI ISHIGO Depatment of Clinical Laboratory Medicine, Ogaki Municipal Hospital MASASHI MIYASATO and ETSUJI ICHIKAWA Meiji Seika Kaisha, Ltd. We analyzed Streptococcus pneumoniae isolates confirmed by direct PCR in Gifu prefecture between May 2002 and August 2002. We analyzed isolates of 254 strains from 6 hospitals to determine antibiotic susceptibility, genotype of penicillin-binding protein (PBP) genes and macrolide resistant genes, and the serotypes distribution of isolates from Matsubara Otorhinolaryngology Clinic. Isolates in which abnormal PBP genes of pbp1a, pbp2x, and pbp2b were identified by PCR were classified based on PCR results as follows; (i) penicillin-susceptible (PSSP) with 3 normal PBP genes, (ii) penicillin-intermediate (PISP) with an abnormal pbp2x, (iii) PISP with an abnormal pbp2b, (iv) PISP with abnormal pbp2x and pbp2b, (v) PISP with abnormal pbp1a and pbp2x, (vi) penicillin-resistant (PRSP) with 3 abnormal PBP genes. The overall incidence of PRSP, PISP and PSSP was 121 (49%), 109 (42%) and 24 (9%), respectively, and there was a significant difference among some hospitals (p 0.05). However, there was no significant difference among the hospitals for the incidence of abnormal macrolide-resistant genes (mefa, ermb). Panipenem showed an excellent antimicrobial activity for injectable carbapenems against PRSP, following biapenem, imipenem, and meropenem. Cefditoren (CDTR) showed an excellent antimicrobial activity for oral cephalosporins against PRSP, following cefteram and cefcapene. Interestingly, there were 2 and 3 strains on MIC of CDTR for 8 and 4 m g/ml, respectively. The prevalent pneumococcal serotypes of isolates in Matsubara Clinic were 6 (17/55), following by 40 (8/55), 9 (6/5) and 15 (5/ 55). The endemic strains were observed in this study using pulsed field gel electrophoresis. These findings suggest the needs to continue the surveillance of bacterial resistance not only in the nationwide but also in the distict.