EV0132 Antimicrobial susceptibility profiles of cinaedi strains isolated from patients in Japan 25thECCMID 2015 Junko Tomida1, Yuji Morita1, Takuichi Sato2, Yoshiaki Kawamura 1 Copenhagen, Denmark 1Department of Microbiology, School of Pharmacy, Aichi Gakuin University, Nagoya, Japan 2Division of Oral Ecology and Biochemistry, Tohoku University Graduate School of Dentistry, Sendai, Japan

Introduction Results and Discussion Table 1. Effects of 16S rRNA gene mutations on PAGU1676-Hospital L A (Hokkaido) PAGU1565-Hospital L PAGU1662-Hospital N 16S rRNA gene sequence •Helicobacter cinaedi is classified as an 8 isolates tetracycline resistance. PAGU1620-Hospital N PAGU1619-Hospital N B (Hokkaido) Japan Number of isolates for which the PAGU1615-Hospital N analysis indicated that enterohepatic Helicobacter species Mutations PAGU1916-Hospital C 1 isolate Tetracycline MIC (mg/mL) PAGU1626-Hospital C H. cinaedi isolates were causing fever, diarrhea, , , on AGA PAGU1286-Hospital A C (Fukushima) 926-928 PAGU1285-Hospital A ≤0.06 0.12 0.25 0.5 1 2 4 PAGU1591-Hospital C divided into 4 clusters proctitis and bacteremia. The number of 20 isolates E (Tokyo) PAGU1516-Hospital C 3 isolates Cluster PAGU1295-Hospital A (Fig. 2). cases of nosocomial infections is growing. D (Chiba) TGC 3 4 4 2 3, 4 PAGU1913-Hospital K 3 isolates F (Tokyo) PAGU1896-Hospital K GTA 1 PAGU1282-Hospital A •There are no recommended guidelines 5 isolates PAGU1283-Hospital A K (Shizuoka) PAGU1279-Hospital A G (Tokyo) ATC 1 PAGU1281-Hospital A H. cinaedi strains with available for susceptibility testing and 9 isolates PAGU1497-Hospital C 1 isolate Cluster PAGU1906-Hospital K L (Aichi) AGC 14 14 16 17 2 PAGU1514-Hospital C double-base-pair mutations treatment of a diagnosed infection by 2 isolates H (Tokyo) 1, 2 PAGU1613-Hospital C 1 isolate GGA 1 3 4 3 PAGU1492-Hospital C on the 16S rRNA (position H. cinaedi. N (Fukuoka) PAGU1616-Hospital N I (Tokyo) PAGU1419-Hospital C 10 isolates AGA 14 2 3 PAGU1764-Hospital M 926-928) showed higher 1 isolate PAGU1617-Hospital N PAGU1909-Hospital K tetracycline MIC values We aimed to assess the PAGU1643-Hospital O J (Tokyo) PAGU1478-Hospital F 11 isolates PAGU1618-Hospital N (MIC ≥ 1 mg/ml) than strains resistance of H. cinaedi isolates Strains of H. cinaedi isolated throughout Japan were PAGU1763-Hospital M M (Osaka) PAGU640-Canada PAGU641-Sweden with single-base-pair or no to various antimicrobial agents, 2 isolates found to be highly resistant to fluoroquinolones. PAGU642-USA O (Kumamoto) H. cinaedi PAGU597T mutations (Table 1). 17 isolates Ciprofloxacin-resistant strains had single or double point PAGU1744-USA -USA in order to improve the PAGU1872-Hospital H PAGU1912-Hospital K Electron microscopic management of H. cinaedi USA 3 isolates mutations in gyrA (Table 2). PAGU1893-Hospital J PAGU1873-Hospital I image of H. cinaedi. Canada 2 isolates PAGU1486-Hospital D infections. Outside Japan PAGU1889-Hospital J Sweden 3 isolates Isolates were resistance to and PAGU1534-Hospital C PAGU611-Hospital O Cluster 1 Australia 1 isolate PAGU1660-Hospital N fluoroquinolones. Penicillins and PAGU1493-Hospital D PAGU1887-Hospital J Fig. 1. Hospital locations in Japan. PAGU1885-Hospital J showed moderate MIC values. Isolates were susceptible PAGU612-Hospital O Materials and Methods PAGU613-Hospital O to and (Table 3). PAGU1749-Sweden PAGU1753-Sweden  Table 2. MICs and amino acid changes in PAGU1884-Hospital J Strains used in this study PAGU1474-Hospital F PAGU1472-Hospital F H. cinaedi strains were isolated in hospitals (A-O) fluoroquinolone-resistant and -susceptible strains of H. cinaedi. PAGU1470-Hospital F PAGU1752-Australia PAGU1406-Hospital D Cluster 2 throughout Japan (Fig. 1). In addition to the No. of MIC (mg/ml) Mutation Fig. 2 Phylogenetic tree based on PAGU1742-Hospital C Isolation PAGU1886-Hospital J Isolates Ciprofloxacin Levofloxacin Moxifloxacin of GyrA PAGU625-Hospital O Japanese isolates we assayed 9 reference strains 16S rRNA gene sequences PAGU1476-Hospital F PAGU1124-Hospital O isolated outside Japan. USA, Canada, Phylogenetic tree based on 16S rRNA gene PAGU1123-Hospital O 7 0.06-0.25 0.06-0.12 0.06 - PAGU1125-Hospital O Mutation on 926-928 Sweden sequences constructed with the neighbor- PAGU1810-Hospital C PAGU629-Hospital E joining method. PAGU1888-Hospital J AGA→TGC group  susceptibility testing Sweden, Australia Thr84Ile PAGU1024-Hospital O 2 16-32 4 0.25 PAGU627-Hospital O Antimicrobial susceptibility was determined by PAGU628-Hospital E PAGU639-Canada Hospital PAGU1614-Hospital N broth microdilution method using modified Thr84Ile Cluster 3 PAGU1784-Hospital B 58 8-64 2-64 0.25-8 PAGU1625-Hospital O A,C,D,E,F,L,N,O PAGU1663-Hospital N levinthal broth. A modified levinthal broth 0.002 PAGU1294-Hospital A Thr84Ile, PAGU1382-Hospital O contained 10 ml of defibrinated horse blood, Hospital C 1 16-31 16 4 PAGU1008-Hospital O Asp88Gly PAGU1894-Hospital J 2+ PAGU1811-Hospital I 1000 ml of mueller hinton broth, 25 mg of Ca , Cluster 4 PAGU1890-Hospital J Thr84Ile, PAGU1892-Hospital J 2+ Hospital C,N 2 32 16 8 PAGU1891-Hospital J and 12.5 mg of Mg (per liter). After heating at Asp88Asn MIT93-1909(U18766) ATCC51401T(AY631945) ATCC49282T 100 ◦C for 5min the modified broth was filter- Thr84Ile, T Hospital C,N,O 12 ≥64 32-64 8-32 Hh-2 (U07574) (M80205) sterilized. Plates were incubated for 72 h at 37 ◦C Asp88Gly, Asn under microaerobic conditions. Table 3. Patterns of primary antibiotic susceptibility of H. cinaedi. MIC50 and MIC90, MICs at which 50 and 90%, respectively, of the isolates are inhibited. Nucleotide sequence analysis Penicillins Cephalosporins Carbapenems Aminoglycosides Tetracycline Macrolides Fluoroquinolones Piperacillin/ Imipenem/ 16S rRNA and gyrA gene sequences were MIC (mg/mL) Ampicillin Amoxicillin Carbenicilin Piperacillin Cefepime Ceftriaxone Meropenem Gentamicin Kanamycin Tetracycline Erythromycin Ciprofloxacin Levofloxacin Moxifloxacin Metronidazole determined. Multiple sequence alignments of Tazobactam Cilastatin DNA sequences and deduced amino acid Range of MIC 0.5-64 1-16 0.5-64 1-8 1-8 0.06-32 1-16 0.06-4 0.06-0.5 0.06-2 0.12-2 0.06-4 8->64 0.06->64 0.06->64 0.06->32 0.06->64

sequences were compared using DNASIS pro MIC50 8 8 8 4 4 4 4 0.5 0.06 0.5 0.5 0.25 >64 32 4 1 16 software. MIC90 16 16 16 8 8 16 8 0.12 0.25 1 1 1 >64 64 64 8 >64