Spodiobacter cordis gen. nov. sp. nov., a member of the family Flavobacteriaceae isolated from patients with infective endocarditis.

Two gram-negative, catalase-negative, oxidase-positive strains (PAGU 1467T and PAGU 1468) isolated from patients with infective endocarditis were investigated to determine their taxonomic status. 16S rRNA gene sequence analysis indicated that the two strains were members of the Bergeyella-Chryseobacterium-Riemerella branch of the family Flavobacteriaceae. Strains PAGU 1467T and PAGU 1468 were highly related to each other (98.8% 16S rRNA gene sequence similarity). Phylogenetically closely-related species to PAGU 1467T comprised Bergeyella zoohelcum (95.0% 16S rRNA gene sequence similarity), Riemerella anatipestifer (94.3%) and Cloacibacterium normanense (94.3%). The major fatty acids of the two isolates were iso-C15:0 , iso-C17:0 3-OH and iso-C15:0 3-OH. The presence of C16:0 3-OH and iso-C15:0 2-OH allowed these isolates to be distinguished from B. zoohelcum. Menaquinone MK-6 was the only respiratory quinone in these organisms; this is a consistent characteristic of the family Flavobacteriaceae. The guanine-plus-cytosine content of the genomic DNA was 42.0%, which is higher than that of other close phylogenetic relatives. On the basis of their phenotypic properties and genetic distinctiveness, isolates PAGU 1467T and PAGU 1468 were classified within the novel genus Spodiobacter, as Spodiobacter cordis gen. nov., sp. nov., which is also the type species. The type strain of S. cordis is PAGU 1467T ( = CCUG 65564T = NBRC 109998T ).

Molecular typing of methicillin-resistant Staphylococcus aureus: Comparison of PCR-based open reading frame typing, multilocus sequence typing, and Staphylococcus protein A gene typing.

The recently developed PCR-based open reading frame typing (POT) method is a useful molecular typing tool. Here, we evaluated the performance of POT for molecular typing of methicillin-resistant Staphylococcus aureus (MRSA) isolates and compared its performance to those of multilocus sequence typing (MLST) and Staphylococcus protein A gene typing (spa typing). Thirty-seven MRSA isolates were collected between July 2012 and May 2015. MLST, spa typing, and POT were performed, and their discriminatory powers were evaluated using Simpson's index analysis. The MRSA isolates were classified into 11, 18, and 33 types by MLST, spa typing, and POT, respectively. The predominant strains identified by MLST, spa typing, and POT were ST8 and ST764, t002, and 93-191-127, respectively. The discriminatory power of MLST, spa typing, and POT was 0.853, 0.875, and 0.992, respectively, indicating that POT had the highest discriminatory power. Moreover, the results of MLST and spa were available after 2 days, whereas that of POT was available in 5 h. Furthermore, POT is rapid and easy to perform and interpret. Therefore, POT is a superior molecular typing tool for monitoring nosocomial transmission of MRSA.

First Japanese case of infectious endocarditis due to Enterococcus faecalis small-colony variants.

A male patient was admitted to our hospital due to infectious endocarditis. He had been treated with levofloxacin for 6 weeks, sulbactam/cefoperazone for 4 weeks, and benzylpenicillin for 2 days prior to valve replacement surgery. Gram-positive cocci, with morphology consistent with γ-Streptococcus, were detected in blood cultures obtained at admission, as well as in vegetation obtained from the aortic valve. However, the strain could not be identified using biochemical methods. Sequencing of the 16S rRNA gene indicated that the culture was a small-colony variant of Enterococcus faecalis. This is the first case in Japan of infectious endocarditis due to E. faecalis small-colony variants. Small-colony variants are subpopulations of bacteria with slow growth, reduced sugar fermentation, and unstable phenotype. As a result, these strains tend to be misidentified. Further, small-colony variants are associated with recurrent and persistent infections such as prosthetic joint infection and infectious endocarditis. These strains are found in various bacterial species, including Staphylococcus aureus, Staphylococcus epidermidis, Escherichia coli, and Pseudomonas aeruginosa, but rarely in Enterococcus species. The case highlights the need to be vigilant of E. faecalis small-colony variants, especially in patients who received long-term courses of antibiotics.

First report of KPC-2 Carbapenemase-producing Klebsiella pneumoniae in Japan.

We investigated a novel Japanese isolate of sequence type 11 (ST11), the Klebsiella pneumoniae carbapenemase-2 (KPC-2)-producing K. pneumoniae strain Kp3018, which was previously obtained from a patient treated at a Brazilian hospital. This strain was resistant to various antibiotic classes, including carbapenems, and harbored the gene blaKPC-2, which was present on the transferable plasmid of ca. 190 kb, in addition to the blaCTX-M-15 gene. Furthermore, the ca. 2.3-kb sequences (ISKpn8-blaKPC-2-ISKpn6-like), encompassing blaKPC-2, were found to be similar to those of K. pneumoniae strains from China.

Catheter-related bloodstream infection by Tsukamurella inchonensis in an immunocompromised patient.

We report a case of catheter-related bloodstream infection by Tsukamurella inchonensis, identified using 16S rRNA gene sequencing, in a patient with myelofibrosis who underwent a bone marrow transplant. Tsukamurella species infections are rare. To our knowledge, this is the first case of T. inchonensis bloodstream infection in an immunocompromised patient.

First report of sepsis caused by Rhodococcus corynebacterioides in a patient with myelodysplastic syndrome.

We report a case of sepsis caused by Rhodococcus corynebacterioides, identified using 16S rRNA gene sequencing, in a myelodysplastic syndrome patient who had undergone hematopoietic stem cell transplantation. This is the first report of R. corynebacterioides infection in a human.

uspA1 of Moraxella catarrhalis clinical isolates in Japan and its relationship with adherence to HEp-2 cells.

The surface protein, UspA1, of Moraxella catarrhalis is involved in adherence to human epithelial cells. We examined the expression of uspA1, and adherence to HEp-2 cells of clinical isolates. The uspA1 gene was detected in 204 of 208 isolates. The 4 uspA1-negative isolates belonged to the 16S rRNA type II with an A to G substitution at nucleotide 445 of 16S rRNA. In 13 isolates of the 16S rRNA type II, transcription of uspA1 was decreased or absent. A relationship between the extent of uspA1 transcription and adherence to HEp-2 cells was found in 5 isolates of the type I. In contrast, the 16S rRNA type II strains still had considerable adherence to HEp-2 cells. The type I uspA1 gene was expressed in Escherichia coli JM109 and the transformants adhered to HEp-2 cells at rates about 7 times higher than the host strain. These data indicated that the uspA1 was virtually ubiquitous in clinical isolates of M. catarrhalis and was responsible for adherence to HEp-2 cells of 16S rRNA type I isolates. However, the data also suggested that adherence of 16S rRNA type II strains to HEp-2 cells was attributed to factor(s) other than UspA1.

Comparison of agar dilution and broth microdilution methods for Clostridium difficile antimicrobial susceptibility testing.

In this study, the performance of the broth microdilution (BMD) method for testing the antimicrobial susceptibility of Clostridium difficile in comparison with the agar dilution (AD) method used by the Clinical and Laboratory Standards Institute (CLSI) was evaluated. In total, 70 non-duplicate C. difficile clinical isolates were used in this study. The minimum inhibitory concentrations (MICs) of clindamycin, moxifloxacin, metronidazole and vancomycin were examined using AD and BMD. The results showed that BMD is acceptable for routine antimicrobial susceptibility testing of C. difficile as its performance was comparable with that of the AD method. In addition, it was noted that metronidazole- and vancomycin-resistant isolates are extremely rare in Japan.

Relapsing Campylobacter coli bacteremia with reactive arthritis in a patient with X-linked agammaglobulinemia.

A patient genetically diagnosed with X-linked agammaglobulinemia repeatedly developed bacteremia due to Campylobacter coli (C. coli) for one year and seven months in spite of immunoglobulin replacement therapy. Throughout the clinical course, C. coli with identical genetic patterns was repeatedly isolated from both blood and stool cultures, thus indicating that the patient had latent intestinal infection. The bacteremia was always accompanied by reactive arthritis. Since the immunoglobulin level was extremely low with severe B cell deficiency, the reactive arthritis must have been induced in a humoral immunity-independent manner. Adding oral minocycline following intravenous meropenem was very effective; the stool cultures became negative and the patient has been well for more than one year without relapse of bacteremia.

Multicenter study to evaluate bloodstream infection by Helicobacter cinaedi in Japan.

Helicobacter cinaedi has being recognized as an important human pathogen which causes bloodstream infections. Although the first case of bacteremia with this pathogen in Japan was reported in 2003, the true prevalence of H. cinaedi as a pathogen of bloodstream infections in this country is not yet known. Therefore, the aim of our study was to assess the incidence of bacteremia with H. cinaedi in Japan. We conducted a prospective, multicenter analysis in 13 hospitals during 6 months in Tokyo, Japan. Among positive blood cultures from 1 October 2003 to 31 March 2004, isolates suspected of being Helicobacter species were studied for further microbial identification. Identification of the organisms was based on their biochemical traits and the results of molecular analysis of their 16S rRNA gene sequences. A total of 16,743 blood culture samples were obtained during the study period, and 2,718 samples (17.7%) yielded positive culture results for coagulase-negative staphylococci. Among nine isolates suspected to be Helicobacter species, six isolates were finally identified as H. cinaedi. The positivity rate for H. cinaedi in blood culture was 0.06% of total blood samples and 0.22% of blood samples with any positive culture results. All patients with bacteremia with H. cinaedi were found to have no human immunodeficiency virus (HIV) infection, but many of them had complications with either malignancy, renal failure, or a history of surgical operation. Therefore, our results suggest that bacteremia with H. cinaedi is rare but can occur in compromised hosts other than those with HIV infection in Japan.

Moraxella catarrhalis does not grow on nutrient agar without sodium chloride supplementation.

None of the 58 Moraxella catarrhalis strains grew on nutrient agar without sodium chloride supplementation, whereas 49 of 51 commensal Neisseria spp. strains tested did. Growth on nutrient agar without sodium chloride supplementation could be used for screening between M. catarrhalis and commensal Neisseria spp.

Isolation of Helicobacter cinaedi from blood of an immunocompromised patient in Japan.

We report the isolation of Helicobacter cinaedi (previously called " Campylobacter-like organism") from the blood of an immunosuppressed Japanese patient receiving immunosuppressive therapy after renal transplantation. The identification of H. cinaedi was based on the findings of microscopic examinations, biochemical properties, and 16S rRNA gene nucleotide sequences. H. cinaedi bacteremia should be considered as one of the critical infectious diseases in immunocompromised patients, and the sequencing of 16S rRNA may be a useful method to confirm the identification of this organism.