The Novel Arylamidine T-2307 Demonstrates In Vitro and In Vivo Activity against Candida auris.

The in vitro and in vivo activity of the arylamidine T-2307 against Candida auris was evaluated. T-2307 demonstrated in vitro activity (MIC ranges ≤ 0.008 to 0.015 µg/ml at 50% inhibition; 0.125 to >4 µg/ml at 100% inhibition). Treatment with T-2307 (3 mg/kg subcutaneous [SC] once daily) also significantly improved survival (70% at 21 days postinfection) and reduced kidney fungal burden (5.06 log10 CFU/g) compared to control (0% survival and 7.09 log10 CFU/g) (P < 0.01).

The Novel Arylamidine T-2307 Selectively Disrupts Yeast Mitochondrial Function by Inhibiting Respiratory Chain Complexes.

The novel arylamidine T-2307 exhibits broad-spectrum in vitro and in vivo antifungal activities against clinically significant pathogens. Previous studies have shown that T-2307 accumulates in yeast cells via a specific polyamine transporter and disrupts yeast mitochondrial membrane potential. Further, it has little effect on rat liver mitochondrial function. The mechanism by which T-2307 disrupts yeast mitochondrial function is poorly understood, and its elucidation may provide important information for developing novel antifungal agents. This study aimed to determine how T-2307 promotes yeast mitochondrial dysfunction and to investigate the selectivity of this mechanism between fungi and mammals. T-2307 inhibited the respiration of yeast whole cells and isolated yeast mitochondria in a dose-dependent manner. The similarity of the effects of T-2307 and respiratory chain inhibitors on mitochondrial respiration prompted us to investigate the effect of T-2307 on mitochondrial respiratory chain complexes. T-2307 particularly inhibited respiratory chain complexes III and IV not only in Saccharomyces cerevisiae but also in Candida albicans, indicating that T-2307 acts against pathogenic fungi in a manner similar to that of yeast. Conversely, T-2307 showed little effect on bovine respiratory chain complexes. Additionally, we demonstrated that the inhibition of respiratory chain complexes by T-2307 resulted in a decrease in the intracellular ATP levels in yeast cells. These results indicate that inhibition of respiratory chain complexes III and IV is a key factor for selective disruption of yeast mitochondrial function and antifungal activity.

Efficacy of T-2307, a novel arylamidine, against ocular complications of disseminated candidiasis in mice.

OBJECTIVES: T-2307, a novel arylamidine, shows broad-spectrum activity against pathogenic fungi, including Candida albicans. Ocular candidiasis is one of the serious complications associated with Candida bloodstream infection and is known to be refractory to conventional antifungal agents. The aim of the present study was to clarify the effectiveness of T-2307 against ocular candidiasis using a mouse model. METHODS: We evaluated ocular fungal burden in mice infected with C. albicans that received treatment with antifungal agents [T-2307, liposomal amphotericin B (LAMB) or fluconazole] for 3 consecutive days. We also assessed survival rates of mice after C. albicans infection followed by treatment for 7 consecutive days. In addition, ocular T-2307 concentrations and in vitro effectiveness against C. albicans biofilm formation were evaluated. RESULTS: The ocular fungal burdens were significantly reduced after T-2307 treatment compared with the control group (no treatment received) and were comparable with those observed following treatment with LAMB or fluconazole in both early- and late-phase treatment experiments. In addition, all of the mice treated with antifungal agents survived for 3 weeks after infection, whereas mice in the control group died within 3 days. The ocular T-2307 trough concentration was maintained above the MIC in the infected mice. An in vitro biofilm inhibition experiment showed that T-2307 suppressed C. albicans biofilm formation at the sub-MIC level, which was comparable with amphotericin B. CONCLUSIONS: Given these results in experimental disseminated candidiasis, T-2307 may be an effective treatment against the complication of ocular candidiasis.

Antifungal susceptibility trend and analysis of resistance mechanism for Candida species isolated from bloodstream at a Japanese university hospital.

We compared the susceptibility of six commercially available antifungal agents (fluconazole, itraconazole, voriconazole, caspofungin, micafungin, and amphotericin B) against 133 Candida bloodstream isolates between 2008 and 2013 at Aichi Medical University Hospital. C. albicans was the most common isolate, followed by C. parapsilosis, C. glabrata, and C. tropicalis. MIC90s of voriconazole against C. albicans, C. parapsilosis, and C. tropicalis were the lowest and that of micafungin against C. glabrata was the lowest among the agents tested. Of the 133 isolates, two strains were identified as drug-resistant. One was a fluconazole-resistant C. glabrata strain, in which the ATP-binding cassette (ABC) transporter gene expression was upregulated. The other was a micafungin-resistant C. glabrata strain, that had 13 amino acid substitutions in FKS1 and FKS2, including a novel substitution V1342I in FKS1 hotspot 2. We also evaluated the susceptibility of T-2307, a novel class of antifungal agents used in clinical trials, against the fluconazole- and micafungin-resistant C. glabrata strain; the MICs of T-2307 were 0.0039 and 0.0078 µg/mL, respectively. In conclusion, the incidence of bloodstream infection caused by drug-resistant Candida spp. was rare from 2008 to 2013 at our hospital. Of 133 isolates, only two strains of C. glabrata were resistant to azoles or echinocandins, that upregulated the ABC transporter genes or had novel FKS mutations, respectively.

In vitro and in vivo antifungal activities of T-2307, a novel arylamidine, against Cryptococcus gattii: an emerging fungal pathogen.

Objectives: T-2307, a novel arylamidine, exhibits potent broad-spectrum activities against the majority of fungal pathogens. In this study, the antifungal activity of T-2307 against Cryptococcus gattii was evaluated in comparison with those of amphotericin B, fluconazole and voriconazole in vitro and in vivo . Methods: The MICs for 15 clinical isolates were determined according to CLSI guidelines and time-kill studies were performed using C. gattii YF2784. In a murine model for intranasal pulmonary infection caused by C. gattii YF2784, the test compounds were administered once daily for 7 days from 2 h or 14 days post-infection. The viable counts in the lungs and brain were determined at 21 days post-infection. Results: The MIC range, MIC 50 , MIC 90 and geometric mean MIC of T-2307 were 0.0078-0.0625, 0.0313, 0.0625 and 0.0394 mg/L, respectively. The MIC of T-2307 was significantly lower than those of fluconazole, voriconazole and amphotericin B. T-2307 showed concentration-dependent fungicidal activity at 4 times the MIC or higher. Administration of T-2307 at 2 mg/kg/day, amphotericin B at 1 mg/kg/day and fluconazole at 160 mg/kg/day from 2 h post-infection significantly reduced viable counts in the lungs and brain. However, when the administration was started 14 days post-infection, only T-2307 significantly reduced the viable counts in both the lungs and the brain at 1 mg/kg/day. Conclusions: T-2307 shows excellent in vitro and in vivo antifungal activities against C. gattii and would be a promising new candidate for the treatment of cryptococcosis.

Impact of the pneumococcal conjugate vaccine on serotype distribution of adult non-invasive Streptococcus pneumoniae isolates in Tokai region, Japan, 2008-2016.

The introduction of the 7-valent pneumococcal conjugate vaccine (PCV7) in February 2010 markedly reduced the burden of invasive pneumococcal disease and changed serotype distribution in Japan. In November 2013, PCV7 was replaced by the 13-valent pneumococcal conjugate vaccine (PCV13). We investigated the serotype distribution and susceptibility trends of non-invasive Streptococcus pneumoniae isolates collected from adult patients. A total of 504 pneumococcal isolates were collected during 4 periods between 2008 and 2016 (Period 1; between June 2008 and April 2009, Period 2; between September 2010 and March 2011, Period 3; between October 2011 and March 2012, Period 4; between August 2015 and January 2016). The coverage of PCV7 and PCV13 significantly decreased from 38.6% and 60.5% in Period 1 to 6.6% and 31.1% in Period 4. This change was mainly due to a large decrease in the frequency of serotype 19F, 6B, and 14. Serotype 3 was the most frequently isolated, and gradually increased. Additionally, non-PCV13 serotypes 11A, 33F, and 35B significantly increased. Most of the PCV7 serotypes 19F, 23F, 6B, and 14 had mutations of penicillin-binding protein genes and macrolide resistance genes, and these serotypes showed low susceptibilities to cefdinir and clarithromycin. On the other hand, a significant change in susceptibility to various antimicrobial agents was not observed throughout the study period, except for decreased susceptibility to carbapenems. Continuous surveillance studies of pneumococcal serotype changes and drug susceptibility are necessary in future.

The novel arylamidine T-2307 demonstrates in vitro and in vivo activity against echinocandin-resistant Candida glabrata.

OBJECTIVES: Candida species are major causes of invasive mycoses in immunocompetent and immunocompromised hosts. Treatment options are limited in the setting of antifungal resistance and increased rates of echinocandin-resistant Candida glabrata have been reported. The novel arylamidine T-2307 demonstrates potent in vitro antifungal activity against Candida species. Our objective was to evaluate the in vitro and in vivo activity of T-2307 against resistant C. glabrata. METHODS: In vitro activity was determined against 42 clinical C. glabrata isolates, including 17 echinocandin-resistant strains. Neutropenic ICR mice were inoculated intravenously with an echinocandin-resistant C. glabrata isolate (T-2307; caspofungin MICs ≤0.008 and 0.5 mg/L, respectively). Therapy with vehicle control, T-2307 (0.75, 1.5, 3 or 6 mg/kg subcutaneously once daily) or caspofungin (1 or 10 mg/kg intraperitoneally once daily) began 1 day post-challenge. Kidneys were collected on day 8 and fungal burden was assessed by counting cfu. RESULTS: T-2307 demonstrated potent in vitro activity against C. glabrata (geometric mean MIC 0.0135 mg/L), which was maintained against echinocandin-resistant isolates (geometric mean MIC 0.0083 mg/L). T-2307 also demonstrated in vivo efficacy in mice infected with echinocandin-resistant C. glabrata. Significant reductions in fungal burden were observed at each dosage level of T-2307 compared with control. Reductions in fungal burden were also observed with high-dose caspofungin. CONCLUSIONS: T-2307 demonstrated potent in vitro activity against C. glabrata, including echinocandin-resistant isolates, which translated into in vivo efficacy against invasive candidiasis caused by an echinocandin-resistant C. glabrata strain. These results demonstrate the potential for T-2307 as therapy against echinocandin-resistant Candida.

T-2307, a novel arylamidine, is transported into Candida albicans by a high-affinity spermine and spermidine carrier regulated by Agp2.

OBJECTIVES: T-2307, a novel arylamidine, exhibits potent broad-spectrum activities against pathogenic fungi, particularly Candida albicans. We previously reported that T-2307 uptake was mainly mediated by a saturable high-affinity carrier at the MIC for C. albicans. Since we hypothesized that the potent anticandidal activity arose from accumulation via the high-affinity carrier, we characterized the specificity and kinetic features of the carrier. METHODS: The MICs of T-2307 for C. albicans strains were evaluated in the presence and absence of potential competitive substrates. The cells were exposed to [(14)C]T-2307, [(14)C]spermine or [(14)C]spermidine in the presence of unlabelled T-2307, pentamidine, propamidine, or competitive substrates if necessary, and the radioactivity in the cells was measured. C. albicans gene deletion was performed using a one-step PCR-based technique. RESULTS: Coapplication with exogenous spermine or spermidine decreased the antifungal activity and uptake of T-2307 in C. albicans strains. T-2307 competitively inhibited spermine and spermidine uptake with inhibition constants similar to its Km for the high-affinity carrier. The comparison of MICs and kinetic values between T-2307 and other diamidine compounds suggested that the different antifungal properties could be partially attributable to the variations in their affinity with the carrier. Studies of gene deletion mutants revealed that T-2307 was transported into C. albicans by a high-affinity spermine and spermidine carrier regulated by Agp2. CONCLUSIONS: Uptake of T-2307 via the high-affinity spermine and spermidine carrier regulated by Agp2 could contribute to its potent antifungal activity. Further investigation is required to identify the high-affinity carrier for potential targeting with novel therapies.

The novel arylamidine T-2307 maintains in vitro and in vivo activity against echinocandin-resistant Candida albicans.

We evaluated the in vitro and in vivo activities of the investigational arylamidine T-2307 against echinocandin-resistant Candida albicans. T-2307 demonstrated potent in vitro activity, and daily subcutaneous doses between 0.75 and 6 mg/kg of body weight significantly improved survival and reduced fungal burden compared to placebo control and caspofungin (10 mg/kg/day) in mice with invasive candidiasis caused by an echinocandin-resistant strain. Thus, T-2307 may have potential use in the treatment of echinocandin-resistant C. albicans infections.

[Sensitivity surveillance of Streptococcus pneumoniae isolates for several antibacterial agents in Gifu and Aichi prefectures (2011-2012)].

We investigated the susceptibility to antibacterial agents, genotype of penicillin-binding protein (PBP) genes and macrolide resistant genes, and the serotypes against 270 strains of Streptococcus pneumoniae isolated from medical facilities in Gifu and Aichi prefectures between October 2011 and April 2012. These results were compared with those against S. pneumoniae isolated in 2008-2009 and 2010-2011. The number of gPSSP with 3 normal PBP genes, gPISP with 1 or 2 normal PBP genes and gPRSP with 3 abnormal genes isolated in 2011-2012 was 15 (5.6%), 162 (60.0%) and 93 (34.4%) strains, respectively. Compared with those isolated in 2008-2009 and 2010-2011, the numbers of gPRSP were decreasing. On the other hand, the isolates with no macrolide-resistant gene, only mefA, only ermB, and both mefA and ermB were 16 (5.9%), 75 (27.8%), 153 (56.7%) and 26 (9.6%). Compared with those isolated in 2008-2009 and 2010-2011, the numbers of isolates with ermB, which was usually associated with high-level resistance, were increasing. The prevalent pneumococcal serotypes in children were type 3 (14.4%), following by type 15 and 19F (9.3%). The coverages of 7-valent pneumococcal conjugate vaccine (PCV7) and 13-valent pneumococcal conjugate vaccine (PCV13) were calculated as 22.9% and 49.2%, respectively. The coverages of PCV7 and PCV13 in gPRSP isolated from children were 47.7% (21/44 strains) and 72.7% (32/44 strains). The MIC90 of each antibacterial agent was as follows; 0.125pg/mL for imipenem, panipenem and garenoxacin, 0.25 µg/mL for meropenem and doripenem, 0.5 µg/mL for cefditoren, moxifloxacin and tosufloxacin, 1 µg/mL for amoxicillin, clavulanic acid/amoxicillin, cefteram, cefcapene and ceftriaxone, 2 µg/mL for benzylpenicillin, ampicillin, sulbactam/ampicillin, piperacillin, tazobactam/piperacillin and levofloxacin, 4 µg/mL for cefdinir, flomoxef and pazufloxacin, 16 µg/mL for minocycline, > 64 µg/mL for clarithromycin and azithromycin, and these MIC90s were about the same as those in 2010-2011.

Characterization of plasmid-mediated quinolone resistance determinants in Klebsiella pneumoniae and Escherichia coli from Tokai, Japan.

The spread of plasmid-mediated quinolone resistance (PMQR) determinants was evaluated in 150 ceftazidime or cefotaxime-resistant clinical isolates of Klebsiella pneumoniae and Escherichia coli from Tokai, Japan between 2008 and 2011. In this study, qnrB, qnrS, and aac(6')-Ib-cr genes were detected in 12 (50.0%), 4 (16.7%), and 1 (4.2%) of 24 K. pneumoniae isolates, respectively, while qnrA, aac(6')-Ib-cr, and qepA genes were detected in 1 (0.8%), 11 (8.7%), and 2 (1.6%) of 126 E. coli isolates, respectively. qnr genes were mainly found in K. pneumoniae (66.7%) and to a lesser extent in E. coli (0.8%). We determined the genetic environment of the qnrB4 gene in 24.6 kb class 1 integron structure, including aar-2, cmlA, blaOXA-10, aadA1, qacEdelta1, sul1, and blaDHA-1. In a time-kill assay, introduction of the qnrB4 or qnrS1 plasmid to the recipient E. coli strain decreased the bactericidal activities of fluoroquinolones such as ciprofloxacin, levofloxacin, and pazufloxacin.

Impact of the pneumococcal conjugate vaccine on serotype distribution and susceptibility trends of pediatric non-invasive Streptococcus pneumoniae isolates in Tokai, Japan over a 5-year period.

Introduction of the 7-valent pneumococcal conjugate vaccine (PCV7) in February 2010 markedly reduced the burden of invasive pneumococcal disease (IPD) and changed serotype distribution in Japan. We investigated the serotype distribution and susceptibility trends of non-invasive Streptococcus pneumoniae isolates collected from pediatric patients. A total of 564 pneumococcal isolates were collected over a 5-year period between 2008 and 2012. The coverage of PCV7 significantly decreased throughout the study period, from 49.3% in period 1 (between June 2008 and April 2009) to 23.4% in period 4 (between October 2011 and March 2012). This change was mainly due to a large decrease in the frequency of 19F (from 20.6% to 9.9%) and 6B (from 10.3% to 2.7%) and an increase in serotype 3 (from 5.1% to 13.5%) and serogroup 15 (from 4.4% to 9.0%). According to serotype replacement, the susceptible ratios of S. pneumoniae to ß-lactams increased slightly while macrolide resistance remained high. The high frequency of macrolide-resistant pneumococcal isolates may continue because of the high frequency of erm(B) in replace serotypes such as serotype 3 and serogroup 15. The continuous surveillance study is essential following the introduction of a second generation 13-valent pneumococcal conjugate vaccine (PCV13).

[Antimicrobial activity of several drugs against extended-spectrum beta-lactamase positive Enterobacteriaceae isolates in Gifu and Aichi prefecture].

We investigated the antimicrobial activity of several drugs against 131 extended-spectrum beta-lactamase (ESBL) positive clinical isolates in Gifu and Aichi prefecture from 2007 to 2011. Meropenem (MEPM) and doripenem (DRPM) gave the lowest MIC50 at 0.0313 microg/mL. MEPM gave the lowest MIC90 at 0.0625 microg/mL. According to the Clinical and Laboratory Standards Institute breakpoints, the susceptible rates of carbapenems, tazobactam/piperacillin (TAZ/PIPC) and cefmetazole (CMZ) were higher than 90%. The susceptible rates of MEPM, DRPM, imipenem (IPM), TAZ/PIPC and CMZ were 98.5%, 98.5%, 94.7%, 94.7% and 92.4%. We used the PCR method and identified the molecular types of the ESBL positive isolates. Seventy-two strains had CTX-M-9 group gene and CTX-M-9 group gene is the most frequently detected. Against the CTX-M-9 group gene harboring strains which were the most common in our investigation, the susceptible rates of TAZ/PIPC, MEPM, DRPM and IPM were 100%. It is suggested that not only carbapenems but also TAZ/PIPC and CMZ are useful against infections caused by ESBL positive isolates.

[Sensitivity surveillance of Streptococcus pneumoniae isolates for several antibacterial agents in Gifu and Aichi prefecture (2010-2011)].

We investigated genotype of penicillin-binding protein (PBP) genes and macrolide resistant genes, the serotypes and the susceptibility to antibacterial agents against 258 strains of Streptococcus pneumoniae isolated from medical facilities in Gifu and Aichi prefectures between January 2010 and March 2011. These results were compared with those against 377 strains of S. pneumoniae isolated in 2008-2009. The number of genotype penicillin-susceptible S. pneumoniae (gPSSP) with 3 normal PBP genes, genotype penicillin-intermediate S. pneumoniae (gPISP) with 1 or 2 normal PBP genes and genotype penicillin-resistant S. pneumoniae (gPRSP) with 3 abnormal genes was 11 (4.3%), 135 (52.3%) and 112 (43.4%) strains, respectively. The isolates with no macrolide-resistant gene, only mefA, only ermB, and both mefA and ermB were 17 (6.6%), 65 (25.2%), 143 (55.4%) and 33 (12.8%). The prevalent pneumococcal serotypes isolated from children were type 19F (18.2%), following by type 6A and 15 (11.7%). The potential coverage of pneumococcal conjugate vaccine (PCV7) was 43.8%. The prevalent pneumococcal serotypes isolated from adults were high in order of type 19F (12.8%), type 6A, 3 and 11 (10.3%), excepting non-typable strains (17.9%), and from elderly persons were type 6B (23.2%) and type 3 (13.4%). The MIC90 of each antibacterial agents was as follows; 0.0625 microg/mL for garenoxacin, 0.125 microg/mL for panipenem, 0.25 microg/mL for imipenem, doripenem, tosufloxacin, 0.5 microg/mL for cefditoren, meropenem, moxifloxacin, 1 microg/mL for amoxicillin, clavulanic acid/amoxicillin, cefteram, cefcapene, ceftriaxone, 2 microg/mL for benzylpenicillin, piperacillin, tazobactam/ piperacillin, pazufloxacin, levofloxacin, 4 microg/mL for cefdinir, flomoxef, 16 microg/mL for minocycline, > 64 microg/mL for clarithromycin, azithromycin and these MIC90s were about the same as those in 2008-2009.

T-2307 causes collapse of mitochondrial membrane potential in yeast.

T-2307, an arylamidine compound, has been previously reported to have broad-spectrum in vitro and in vivo antifungal activities against clinically significant pathogens, including Candida species, Cryptococcus neoformans, and Aspergillus species, and is now undergoing clinical trials. Here we investigated the mechanism of action of T-2307 using yeast cells and mitochondria isolated from yeast and rat liver. Nonfermentative growth of Candida albicans and Saccharomyces cerevisiae in glycerol medium, in which yeasts relied on mitochondrial respiratory function, was inhibited at 0.001 to 0.002 µg/ml (0.002 to 0.004 µM) of T-2307. However, fermentative growth in dextrose medium was not inhibited by T-2307. Microscopic examination using Mitotracker fluorescent dye, a cell-permeant mitochondrion-specific probe, demonstrated that T-2307 impaired the mitochondrial function of C. albicans and S. cerevisiae at concentrations near the MIC in glycerol medium. T-2307 collapsed the mitochondrial membrane potential in mitochondria isolated from S. cerevisiae at 20 µM. On the other hand, in isolated rat liver mitochondria, T-2307 did not have any effect on the mitochondrial membrane potential at 10 mM. Moreover, T-2307 had little inhibitory and stimulatory effect on mitochondrial respiration in rat liver mitochondria. In conclusion, T-2307 selectively disrupted yeast mitochondrial function, and it was also demonstrated that the fungal mitochondrion is an attractive antifungal target.

In vitro and in vivo antimalarial activities of T-2307, a novel arylamidine.

T-2307, a novel arylamidine, has been shown to exhibit broad-spectrum antifungal activities against clinically significant pathogens. Here, we evaluated the in vitro and in vivo antimalarial activity of T-2307. The 50% inhibitory concentrations (IC50s) of T-2307 against Plasmodium falciparum FCR-3 and K-1 strains were 0.47 and 0.17 µM, respectively. T-2307 at 2.5 to 10 mg/kg of body weight/day exhibited activity against blood stage and liver stage parasites in rodent malaria models. In conclusion, T-2307 exhibited in vitro and in vivo antimalarial activity.

In vitro antichlamydial activity of garenoxacin against Chlamydia trachomatis.

Garenoxacin showed the most potent chlamydial activity against Chlamydia trachomatis D/UW-3/Cx among three tested quinolones and azithromycin. The DNA gyrase genes, gyrA and gyrB, of C. trachomatis D/UW-3/Cx were cloned and the GyrA and GyrB subunits of DNA gyrase protein were separately expressed as histidine-tagged proteins in Escherichia coli. The mean 50% inhibitory concentration (IC(50)) of garenoxacin against the supercoiling activity of C. trachomatis D/UW-3/Cx gyrase was 2.9 ± 0.4 µg/ml, which was the most potent inhibitory activity against DNA gyrase among the quinolones tested in this study. At an extracellular concentration of 0.5 µg/ml, the cellular-to-extracellular concentration ratio of garenoxacin was 15.3 ± 1.3, equivalent to that of moxifloxacin and greater than that of levofloxacin. In a time-kill experiment, after exposure to garenoxacin at a concentration of 0.5 µg/ml at 0-6, 5-11, and 24-30 h after infection, the percentages of recoverable chlamydial inclusion-forming units were 11.1 ± 3.3, 0.6 ± 0.1, and 2.6 ± 0.5%, respectively. On transmission electron microscopy observation, after exposure to garenoxacin at 24-30 h after infection, some C. trachomatis elementary bodies remained in the inclusion body; however, the reticulate bodies were completely disrupted. In conclusion, garenoxacin is expected to be a useful quinolone in the treatment of infectious diseases caused by C. trachomatis.

[Antibacterial susceptibility surveillance of Haemophilus influenzae isolated from pediatric patients in Gifu and Aichi prefectures (2009-2010)].

We investigated the susceptibility to antibacterial agents of 197 strains of Haemophilus influenzae isolated from pediatric patients at medical facilities in Gifu and Aichi prefectures between 2009 and 2010. Those strains were also examined for the mutations of ftsI coding for penicillin-binding protein 3, presence of bla TEM-1, serotype and beta-lactamase producing ability. Among the 197 strains, the most prevalent serotype was non-typeable (89.8%), followed by serotype b (8.1%), e (1.5%) and f (0.5%). Based on the susceptibility among the 197 strains to antibacterial agents, beta-lactamase nonproducing ampicillin-susceptible H. influenzae (BLNAS) accounted for 27.4%, beta-lactamase nonproducing ampicillin-resistant H. influenzae (BLNAR) for 62.4%, beta-lactamase producing ampicillin-resistant H. influenzae (BLPAR) for 6.1% and beta-lactamase producing amoxicillin/ clavulanic acid-resistant H. influenzae (BLPACR) for 4.1%. According to PCR-based genotyping, the strains were classified into 6 categories: gBLNAS, gLow-BLNAR, gBLNAR, gBLPAR, gBLPACR-I and gBLPACR-II. The incidences of each resistant class were 17.3% for gBLNAS, 6.6% for gLow-BLNAR, 66.0% for gBLNAR, 5.6% for gBLPAR and 4.6% for gBLPACR-II. The combined incidence of gLow-BLNAR and gBLNAR was 72.6%, which was higher than that of BLNAR (62.4%). The MIC90s of antibacterial agents against the 197 strains were as follows; 0.0156 microg/mL for tosufloxacin and garenoxacin, 0.0313 microg/mL for levofloxacin and pazufloxacin, 0.0625 microg/mL for norfloxacin, 0.25 microg/mL for tazobactam/piperacillin (TAZ/PIPC) and ceftriaxone, 0.5 microg/mL for TAZ/PIPC (1:8) and cefditoren, 1 microg/mL for piperacillin, cefteram, cefotaxime, meropenem, tebipenem and minocycline, 2 microg/mL for doripenem, 4 microg/mL for cefcapene, imipenem and azithromycin, 8 microg/mL for sulbactam/ampicillin, clavulanic acid/amoxicillin (1:2, CVA/AMPC) and cefdinir, 16 microg/mL for CVA/AMPC (1:14), flomoxef and clarithromycin, 32 microg/mL for ampicillin. Although there was no rapid increase in the antibacterial resistance, the prevalence of BLNAR was still over 50%. In order to ensure the appropriate chemotherapy, it is important to continue the surveillance of susceptibility among H. influenzae.

[Sensitivity surveillance of Streptococcus pneumoniae isolates for several antibacterial agents in Gifu and Aichi prefecture (2008-2009)].

We investigated the susceptibility to antibacterials, genotype of penicillin-binding protein (PBP) genes and macrolide resistant genes, and the serotypes against 377 strains of Streptococcus pneumoniae isolated from medical facilities in Gifu and Aichi prefectures between June 2008 and April 2009. These results were compared with those against 160 strains of S. pneumoniae isolated in 2004. Referring to CLSI (M100-S17), the overall incidence of penicillin-susceptible (PSSP), penicillin-intermediate (PISP) and penicillin-resistant (PRSP) S. pneumoniae was 143 (38%), 185 (49%) and 49 (13%) strains, respectively. PISP and PRSP were isolated higher in the material of nasal cavity and throat, and PRSP was isolated higher in the area of Chuno district. The number of gPSSP with 3 normal PBP genes, gPISP with 1 or 2 normal PBP genes and gPRSP with 3 abnormal genes was 23 (6.1%), 173 (46%) and 181 (48%) strains, respectively. The isolates with no macrolide-resistant gene, only mefA, only ermB, and both mefA and ermB were 28 (7.4%), 138 (37%), 166 (44%) and 45 (12%). The prevalent pneumococcal serotypes were type 19 (92 strains; 24%), following by type 23 (60 strains; 16%) and type 6 (56 strains; 15%). The 80% of pneumococcal serotypes of PRSP were serotype 19 and 6. The MIC90 of each antibacterial was as follows; 0.1 microg/mL for imipenem, panipenem and garenoxacin, 0.2 microg/mL for moxifloxacin, 0.39 microg/mL for meropenem and tosufloxacin, 0.78 microg/ mL for amoxicillin, clavulanic acid/amoxicillin, cefditoren and cefcapene, 1.56 microg/mL for benzylpenicillin, piperacillin, cefteram and levofloxacin, 3.13 microg/mL for cefotiam, flomoxef and pazufloxacin, 6.25 microg/mL for cefdinir, 12.5 microg/mL for norfloxacin and minocycline, > 100 microg/mL for clarithromycin, and these MIC90s were about the same as those in 2004.

[Sensitivity surveillance of Pseudomonas aeruginosa isolates for several antibacterial agents in Gifu and Aichi prefecture (2008)].

We investigated the susceptibility to antibacterial agents of 334 strains of Pseudomonas aeruginosa isolated from medical facilities in Gifu and Aichi prefectures from May to September 2008. For the beta-lactams, meropenem (MEPM) and doripenem (DRPM) gave the lowest MIC50 at 0.5 microg/mL, and tazobactam/piperacillin (TAZ/PIPC) gave the highest susceptible rate of the breakpoint by Clinical and Laboratory Standards Institute (CLSI) at 93.1%. For the quinolones, ciprofloxacin (CPFX) gave the lowest MIC50 at 0.25 microg/mL, followed by pazufloxacin (PZFX) at 0.5 microg/mL, and levofloxacin (LVFX) at 1 microg/mL, and susceptible rate was 76.0% for CPFX and 73.4% for LVFX. Susceptible rates to amikacin (AMK) and tobramycin (TOB) of aminoglycocides and colistin (CL) of polypeptides were 98.2%, 97.6% and 96.4%. In 334 strains, IMP-1 MBL producing P. aeruginosa was 1 strain, and the strain showed resistance to all antibacterial agents except AMK and CL used in this study. The strains isolated from urine were lower susceptible rate in comparison with those from sputum, notably the susceptible rate to CPFX from urine was less over 30% than those from sputum. Because the results of the susceptibility test against P. aeruginosa were different in each area, it is important for us to pay attention to the susceptibility to antibacterial agents and the emergence of resistance in the clinical strains through continuous susceptibility surveillance.