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We present two cases of bacteremia caused by Leptotrichia trevisanii: a 12-year-old girl with recurrent myeloid leukemia of the mandible and a 66-year-old man with esophageal carcinoma. As this filamentous bacillus showed indefinite Gram staining and the identification based on biochemical enzymatic reactions was not definitive, identification required 16s rRNA analysis. For this organism, drug sensitivity testing showed susceptiblity to each β-lactam antibiotics and clindamycin, but resistance to fluoroquinolone and erythromycin. This filamentous bacillus needs careful identification and appropriate antibiotic treatment.

Introduction: Abiotrophia species have been referred to as nutritionally variant streptococci because of their fastidious nutritional requirements for growth. Abiotrophia species are difficult to identify with conventional solid culture., Presentation of Case: A 48-year-old woman was admitted to our hospital with severe low back pain and body temperature of 38.2°C. Magnetic resonance imaging revealed edema and contrast enhancement of the L4 and L5 vertebral bodies with high signal intensity in the L3-4 and L4-5 intervertebral discs on the T2-weighted images. The patient underwent needle biopsy of the L3-4 disk. Cultures of disk biopsy samples and blood yielded gram positive cocci in short chains with scanty growth on chocolate agar. Further subculture with supplemented medium and subsequent 16S ribosomal RNA gene sequencing identified the pathogen as Abiotrhophia adiacens. The patient was treated with intravenous ampicillin. At 6-month follow-up, the patient was free of symptoms., Discussion: Causative microorganisms remain unidentified in 25-40% of spinal infection cases. Abiotrophia species grow poorly on conventional solid media, and require pyridoxal or thiol group supplementation. Use of Brucella HK agar or GAM agar plate is helpful for detection of Abiotrophia species. We first confirmed the diagnosis by direct identification of Abiotrophia adiacens from infected disk. Abiotrophia species are one of the major pathogens of infective endocarditis accounting for 5% of cases. Considering their fastidious nature, it is likely that most cases of Abiotrophia discitis are falsely classified as culture-negative discitis; therefore, their role in pyogenic discitis may be underestimated., Conclusion: Subculture using nutritionally supplemented media is crucial for their identification., (Copyright © 2013 The Authors. Published by Elsevier Ltd.. All rights reserved.)

To better place the form and motion of the human foot in an evolutionary context, understanding how foot motions change when quadrupeds walk bipedally can be informative. For this purpose, we compared the pressures beneath the foot during bipedal and quadrupedal walking in Japanese macaques (Macaca fuscata). The pressure at nine plantar regions was recorded using a pressure mat (120 Hz), while the animals walked on a level walkway at their preferred speeds. The results revealed substantial differences in foot use between the two modes of locomotion, and some features observed during bipedal walking resembled human gait, such as the medial transfer of the center of pressure (COP), abrupt declines in forefoot pressures, and the increased pressure beneath the hallux, all occurring during the late-stance phase. In particular, the medial transfer of the COP, which is also observed in bonobos (Vereecke et al.: Am J Phys Anthropol 120 (2003) 373-383), was due to a biomechanical requirement for a hind limb dominant gait, such as bipedal walking. Features shared by bipedal and quadrupedal locomotion that were quite different from human locomotion were also observed: the heel never contacted the ground, a foot longitudinal arch was absent, the hallux was widely abducted, and the functional axis was on the third digit, not the second.

Most primates use diagonal sequence (DS), diagonal couplets (DC) gaits when they walk or run quadrupedally, and it has been suggested that DSDC gaits contribute to stability in their natural arboreal habitats compared to other symmetrical gaits. However, this postulate is based solely on studies of primate gaits using continuous terrestrial and arboreal substrates. A particular species may select suitable gaits according to the substrate properties. Here, we analyzed the gaits of Japanese macaques moving on a horizontal ladder with rung intervals ranging from 0.40 to 0.80 m to elucidate the relative advantages of each observed form of gait. The rung arrangement forced our macaques to choose either diagonal coupling or DS gaits. One macaque consistently used diagonal coupling (i.e., DSDC and LSDC gaits) across narrow and intermediate rung intervals, whereas the other macaque used DS gaits (i.e., DSDC and DSLC gaits). At wider rung intervals, both macaques shifted to a two-one sequence (TOS), which is characterized by two nearly simultaneous touchdowns of both forelimbs and one touchdown of each hind limb in a stride. The transition to the TOS sequence increased the duration of support on multiple limbs, but always included periods of a whole-body aerial phase. These results suggest that Japanese macaques prefer DSDC gaits, because the diagonal coupling and DS contribute separately to stability on complex supports compared to the lateral coupling and lateral sequence. We also postulate that stability triggers the transition from symmetrical gaits to the TOS sequence.

Purpose: When smear test is positive for acid-fast bacilli, it is important to differentiate between Mycobacterium tuberculosis complex and nontuberculosis mycobacteria (NTM) in healthcare associated infection (HAI) control. The aim of this study is to evaluate between COBAS TaqMan48 (TaqMan) and COBAS Amplicor (COBAS)., Material and Method: Tenfold dilution series of 5 x 10(4) cfu/mL of Mycobacterium bovis (ATCC19210) were used for evaluating the limit of detection (LOD) and reproducibility. 73 frozen clinical specimens (34 M. tuberculosis complex and 39 NTM) stored at below -20 degrees C before its use that were treated with NALC-NaOH were used to determine the agreement between two methods. Divided reaction reagents (include Master mix solution, Internal control and Mg2+) of TaqMan for 3 and 6 tests per batch used to evaluate reagents stability., Result: The LOD of both kits were 5 X102 cfu/mL. Regarding reproducibility, the same result was obtained when tested 3 times. The agreement rate between TaqMan and culture method was 58.8%, and between COBAS and culture method was 67.6%. When limited to smear positive eleven specimens, the agreement between TaqMan and culture method was 81.8%, and between COBAS and culture method was 90.9%. Reagents divided for 3 tests and 6 tests and stored at 4 degrees C in dark, both test reagents stability was confirmed maximum for 16 days., Conclusion: As the results of the evaluation of TaqMan, the LOD, reproducibility and the agreement were similar to COBAS results. However, in low colony forming unit of clinical specimen raise the possibility that results may contain false negative.

We examined the correlation between the density of muscle spindles in shoulder muscles and the locomotor mode in three species of prosimian primates: the slow loris (Nycticebus coucang), Garnett's galago (Otolemur garnettii), and the ring-tailed lemur (Lemur catta). The shoulder muscles (supraspinatus, infraspinatus, teres major, teres minor, and subscapularis) were embedded in celloidin and cut into transverse serial thin sections (40 microm); then, every tenth section was stained using the Azan staining technique. The relative muscle weights and the density of the muscle spindles were determined. The slow loris muscles were heavier and had sparser muscle spindles, as compared to Garnett's galago. These features suggest that the shoulder muscles of the slow loris are more adapted to generating propulsive force and stabilizing the shoulder joint during locomotion and play a less controlling role in forelimb movements. In contrast, Garnett's galago possessed smaller shoulder muscles with denser spindles that are suitable for the control of more rapid locomotor movements. The mean relative weight and the mean spindle density in the shoulder muscles of the ring-tailed lemur were between those of the other primates, suggesting that the spindle density is not simply a consequence of taxonomic status., (Copyright 2007 S. Karger AG, Basel.)

We studied changes in toxin-producing genes and drug susceptibility in Staphylococcus aureus isolated from blood cultures at the University of Tokyo Hospital between 1980-1984 (six mecA gene-positive methicillin resistant S. aureus [MRSA] strains and 20 mecA gene-negative methicillin-susceptible S. aureus [MSSA] strains) and 1999 (11 MRSA and 20 MSSA strains). The prevalence of strains with toxin-producing genes increased from 66.7% to 90.9% in MRSA, and from 30.0% to 55.0% in MSSA during the interval. Among toxin-producing gene-positive S. aureus, the dominant strains shifted from those with the enterotoxin (ET) - A gene in 1980-1984 to those with both the toxic shock syndrome toxin-1 and the ET-C genes in 1999. All strains were susceptible to vancomycin and teicoplanin. Mupirocin and arbekacin inhibited all strains at concentrations of less than or equal to 0.5 micro g/ml and 4 micro g/ml, respectively. More than half of the MRSA strains in 1999 were considered to be nonsusceptible to flomoxef. Because almost all MRSA and more than half of MSSA among recently isolated strains possessed the toxin-producing genes, we should pay attention to whether toxin-related diseases caused by MRSA and MSSA are increasing.

Amplicor Mycobacterium Kit (Roch Diagnostics: Japan) is the most widely used kit in Japan for the diagnosis of mycobacteria infections, especially those caused by Mycobacterium tuberculosis, Mycobacterium avium and Mycobacterium intracellulare. We evaluated the reliability of the kit in co-operation with 331 laboratories using the kit in routine examination. We distributed specially prepared 4 test samples to each laboratories. The negative sample was NALC-NaOH treated sputum which showed "negative" when tested by this kit and positive samples were NALC-NaOH treated sputum containing M. bovis or M. intracellurare. False-positive results were reported in 6 out of 331 laboratories (1.8%) and false-negative results were reported from 7 laboratories (2.1%). (The details were 1 out of 331 labs for TB-H sample, 5 out of 331 labs for TB-L sample and 1 out of 316 in MIN sample.) Statistical significance between MWP method and COBAS method was not significant. After receiving and evaluating the test results on the 4 samples, the follow up questionnaires were sent out to 22 laboratories which reported incorrect results and low optical density (O.D.) on positive control. Results of this questionnaire suggested that it was important to follow the package insert instructions and to follow the correct procedures for PCR assay. These results suggested that Amplicor Mycobacterium Kit is reliable for rapid diagnosis of Mycobacteria infections.

Since methicillin-resistant Staphylococcus aureus(MRSA) is resistant not only to methicillin but also to multiple species of antibacterial drugs, there are cases where infections with MRSA are difficult to treat. For laboratory identification of MRSA, S. aureus is first identified, then an oxacillin (MPIPC) sensitivity test, PCR detection of the S. aureus mecA gene and an agglutination test using the latex sensitized with an anti-PBP-2' monoclonal antibody are usually utilized. However, the detection of MRSA does not necessarily indicate MRSA infection, and many cases only demonstrate MRSA colonization. Thus, a clinical investigation is indispensable for establishing the diagnosis of MRSA infection. In addition, recent reports also suggested a current trend toward decreasing sensitivity to Bactroban(MUP, mupirocin, an anti-intranasal MRSA antibacterial drug). Taking these into consideration, surveillance of the trend in patient's sensitive to various species of antibacterial drugs including MUP must continue further.

We experienced two Burkholderia cepacia outbreaks over a 1-year period. During this period, 28 B. cepacia isolates were obtained from clinical specimens, and 2 were obtained from environmental specimens (i.e., from a nebulizer solution and a nebulizer tube). These 30 isolates were subjected to the PCR-based randomly amplified polymorphic DNA (RAPD) assay as well as to pulsed-field gel electrophoresis (PFGE). In the first outbreak, in which eight patients hospitalized in the Trauma and Critical Care Center were involved, the RAPD assay revealed that all 20 isolates obtained from clinical specimens and the 2 isolates from environmental specimens had identical DNA profiles. These RAPD data enabled us to pinpoint a possible source and to take countermeasures to prevent further spread of the epidemic-causing strain. In the second outbreak, two consecutive B. cepacia infection/colonization cases were seen in the surgery ward. The RAPD profiles of four isolates obtained were again identical, but they were distinct from those seen in the first outbreak, clearly indicating that the second outbreak was not related to the first. Thus, our experience demonstrated that the RAPD assay is a useful and reliable tool for epidemiological studies of B. cepacia isolates from nosocomial outbreaks. Since the RAPD assay could provide discriminatory potential and reproducibility comparable to those of the widely used PFGE assay with less complexity and in a shorter time, the introduction of the RAPD assay into hospital microbiology laboratories as a routine technique may help prevent nosocomial outbreaks.

The in vitro antibacterial activity of cefozopran (CZOP) against recent clinical isolates was evaluated and compared with those of ceftazidime (CAZ), cefpirome (CPR), cefepime (CFPM), cefotaxime (CTX), sulbactam/cefoperazone (S/C), imipenem (IPM), oxacillin (MPIPC), and flomoxef (FMOX). MIC80 values of CZOP for methicillin-susceptible Staphylococcus aureus (MSSA, n = 41), methicillin-resistant Staphylococcus aureus (MRSA, n = 57), Streptococcus pneumoniae (n = 45), Enterococus faecalis (n = 49), Enterobacter cloacae (n = 50), Citrobacter freundii (n = 45), Serratia marcescens (n = 45), and Pseudomonas aeruginosa (n = 100) were 1, 32, 2, 16, 4, 1, 0.25, 8 micrograms/ml, respectively. CZOP was more active than CPR against P. aeruginosa and exhibited similar activity to CPR against other species. CZOP was especially active against S. marcescens with MIC values lower than 1 microgram/ml against all strains tested. CZOP was similarly active to or more active than CFPM against all species except for C. freundii. CZOP was not active against MRSA. Thus, we investigated the in vitro combination effects of CZOP/vancomycin (VCM) and CZOP/arbekacin (ABK) using the checkerboard method. The interaction between CZOP and VCM ranged from additive activity (0.5 < FIC index < or = 1.00, n = 37) to synergistic activity (FIC index < 0.50, n = 1), except for one strain showing indifference (1.00 < FIC index < or = 2.00). The interaction between CZOP and ABK ranged from additive activity (n = 22) to synergistic activity (n = 1). These date suggest the potential effect of combination therapy of (CZOP) and VCM or ABK against MRSA. The combined therapy is suggested to be useful to reduce side effects in patients with impared renal function, to reduce the administration dose of VCM or to treat infections of sites where achievable drug concentrations are lower than those commonly achieved in the bloodstream. We also investigated the combination effects of CZOP/AMK and CZOP/GM against CZOP-resistant P. aeruginosa (MIC > 16 micrograms/ml). The combination of CZOP/AMK showed additive activity (n = 9) to synergistic activity (n = 2). The combination of CZOP/GM showed additive activity (n = 5). These results suggest that combinations of CZOP with AMK or GM are effective in treating P. aeruginosa.