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As the reagent that can simultaneously detect bacterial nucleic acid/drug-resistant genes from the culture-positive liquid of blood cultures, Verigene ® system includes the Verigene ® Gram-Positive Blood Culture test (BC-GP) and the Verigene ® Gram Negative Blood Culture test (BC-GN). This study used BC-GN to identify the names of bacteria from stock strains, urine samples, and sputum specimens and detect drug-resistant genes. The stock strains included 28 clinical isolates, 9 urine samples in which the target bacterial strain grew to 106CFU/ml or more in culture, and 9 sputum specimens in which the target bacterial strain grew to 105CFU/ml or more in culture. The bacterial identification and detection of drug-resistant genes used quality Matrix Assisted Laser Desorption/Ionization Time of Flight Mass Spectrometry (MALDI-TOF MS) analysis and conventional PCR method, respectively, followed in comparison with the results of Verigene ® . As a result, the measurement results of Verigene ® for the stock strains and urine samples had a high concordance rate with MALDI-TOF MS analysis and PCR method. On the other hand, the concordance rate of the sputum specimens with the Verigene ® measurement results was only 40% (4 out of 10 specimens). These results suggest that BC‒GN can be an effective tool for AMR rapid diagnosis if the measurement target includes not only bacterial strains in the culture-positive liquid of blood cultures, but also other bacterial strains and urine.

A 79-year-old woman was admitted emergently for disturbance of consciousness. Her consciousness level was Japan coma scale 20, and she presented with hypermyotonia. Brain magnetic resonance imaging and cerebrospinal fluid examination showed normal findings. Her blood tests showed an increased ammonia level of 291 μg/dl with normal liver function. We catheterized the bladder for urinary retention. Eight hours after admission, the blood level of ammonia decreased to 57 μg/dl and the patient's consciousness level improved. Corynebacterium pseudodiphtheriticum, which is a bacteria producing urease, was detected from a urine culture. It is important to recognize that obstructive urinary tract infection caused by urease-producing bacteria can cause hyperammonemia.

Twenty-four patients with cutaneous ureterostomy at least 6 months after surgery were included in this study. Single-J ureteral catheters were indwelt in 7 patients. Pouch urine was cultured and analyzed from asymptomatic patients and from those with pyelonephritis. Forty-eight bacterial strains were isolated from asymptomatic patients. Thirty-five of these strains consisting of 18 species were isolated from the patients without ureteral catheters. Enterococcus (E.) faecalis (20.0%), Klebsiella pneumoniae (11.4%) and Proteus (P.) mirabilis (11. 4%) were isolated at high frequency. One methicilline-resistant coagulase-negative Staphylococcus (MRCNS) strain was isolated. Thirteen bacterial strains consisting of 8 species were isolated from the patients with ureteral catheters. E. faecalis (23.1%), Eschericia (E.) coli (23.1%) and Pseudomonas (P.) aeruginosa (15.4%) were isolated at high frequency. One E. coli strain harboring extended-spectrum beta lactamase (ESBL) and one MRCNS strain were isolated. Pyelonephritis occurred in 5 patients 14 times during follow-up for at least 6 months after surgery. The patients with ureteral catheters suffered from pyelonephritis (4/7, 57.1%) more frequently as compared to the patients without catheters (1/17, 5.9%) (p = 0.014). P. aeruginosa (26.9%), E. faecium (15.4%), and P. mirabilis (11.5%) were isolated inhigh frequency. One E. coli strain harboring ESBL and one P. mirabilis strain harboring ESBL were isolated. In conclusion, insertion of ureteral catheters in a cutaneous ureterostomy is a strong risk factor for pyelonephritis and development of antibiotic-resistant bacteria. Therefore, urine culture is important in these patients who are under surveillance.

A 62-year-old man, with a family history of prostate cancer, referred to our hospital because of elevated prostate-specific antigen (PSA) (6.02 ng/ml). After prophylactic administration of antibiotics (cefotiam), transrectal needle biopsy of the prostate was performed. He was admitted to the hospital due to high fever the next evening. His blood pressure was below the shock level, and his renal function deteriorated progressively. Suspecting septic shock, the patient was treated with Meropenem, γ-globulin, and dopamine, which were not effective. Then, endotoxin adsorption therapy was employed and the condition of the patient recovered soon after the initiation of the therapy. Extended spectrum β -lactamase-producing Escherichia coli was found in his urine. Pathological diagnosis of the biopsy specimen was atypical glands.

The antibacterial activity of meropenem (MEPM) and other parenteral antibiotics against clinical isolates of 2655 strains including 810 strains of Gram-positive bacteria, 1635 strains of Gram-negative bacteria, and 210 strains of anaerobic bacteria obtained from 30 medical institutions during 2009 was examined. The results were as follows; (1) MEPM was more active than the other carbapenem antibiotics tested against Gram-negative bacteria, especially against enterobacteriaceae and Haemophilus influenzae. MEPM was also active against most of the species tested in Gram-positive and anaerobic bacteria, except for multidrug resistant strains including methicillin-resistant Staphylococcus aureus (MRSA). (2) MEPM maintained potent and stable antibacterial activity against Pseudomonas aeruginosa. The proportion of MEPM-resistant strains to ciprofloxacin-resistant strains or imipenem-resistant strains were 53.1% and 58.0% respectively. (3) The proportion of extended-spectrum beta-lactamase (ESBL) strains was 3.1% (26 strains) in enterobacteriaceae. And the proportion of metallo-beta-lactamase strains was 2.0% (6 strains) in P. aeruginosa. (4) Of all species tested, there were no species except for Bacteroides fragilis group, which MIC90 of MEPM was more than 4-fold higher than those in our previous study. Therefore, there is almost no significant decrease in susceptibility of clinical isolates to meropenem. In conclusion, the results from this surveillance study suggest that MEPM retains its potent and broad antibacterial activity and therefore is a clinically useful carbapenem for serious infections treatment at present, 14 years passed after available for commercial use in Japan.

Recently, there have been reports concerning an increased frequency of isolation of multi-drug resistant Pseudomonas aeruginosa (MDRP) strains in hospitals and other clinical settings as well as the associated risk of their hospital-acquired infections; in such a situation, it has been a major challenge to establish methods of managing and treating the infections. In order to investigate the trend of P. aeruginosa, the Infection Forum in the Chugoku Region has conducted to a multi-center collaborative study to isolate P. aeruginosa strains from sputum and urine samples collected between October 2006 and September 2008, analyzed the drug susceptibility and the pulsed-field gel electrophoresis (PFGE) patterns of each strain, and assessed epidemiologic characteristics. Of the 738 P. aeruginosa strains collected in this study, 152 (20.6%), 179 (24.3%), 47 (6.4%), and 39 (5.3%) were found to be ciprofloxacin-resistant, imipenem-resistant, amikacin-resistant, and MDRP, respectively. Among the various antimicrobial agents tested, arbekacin (ABK) revealed the strongest inhibitory effects on each drug-resistant and MDRP strain; therefore, ABK was considered as a potential candidate for future treatment of diseases caused by P. aeruginosa. The study also showed that the detection rates of MDRP varied a lot from hospital to hospital. In addition, PFGE-based cluster analyses revealed several strains isolated in the same hospital exhibited a similar PFGE pattern and the same drug susceptibility, suggesting the presence of "unique" hospital-specific strains.

We evaluated performance of Abbott RealTime CT/NG assay (real-time PCR, Abbott Japan) for detect Chlamydia trachomatis and Neisseria gonorrhoeae by real-time PCR in 88 female patients with cervicitis symptoms seen at gynecological clinics and 100 male patients with urethritis symptoms seen at urological or dermatology clinics in Kitakyushu, Japan. Endocervical swab and first-voided urine (FVU) specimens were then collected from women and FVU specimens from men. Detection rates of C. trachomatis and N. gonorrhoeae by real-time PCR in the 3 types of specimens were compared to those by ProbeTec ET assay (ProbeTec, BD Diagnostic System). The overall positive concordance between real-time PCR and ProbTec were 97.1% (66/68) for C. trachomatis and 100% (33/33) for N. gonorrhoeae, C. trachomatis detection yielded 3 discordant results in endocervical specimens and 1 discordant result in male FVU by real-time PCR and ProbTec. Three of 4 reexamined using Aptime Combo 2 Assay (Fuji Rebio Inc.) were positive for C. trachomatis. Endocervical swab and FVU specimen results for C. trachomatis were discordant in 3 cases in real-time PCR and 4 in ProbeTec. Subjects with 2 or more positive endocervical awab results in female or male FVU specimens were assumed to be "true positive" for C. trachomatis. The sensitivities of real-time PCR for detecting C. trachomatis was 94.4% in endocervical swabs, 77.8% in female FVU and 97.4% in the male FVU. The sensitivities for real-time PCR for detecting N. gonorrhoeae was 100% in all 3 specimentypes. Abbott RealTime CT/NG assay was useful for detecting C. trachomatis using endocervical swabs or male FVU specimens and for detecting N. gonorrhoeae using endocervical swabs and all FVU specimens.

We evaluated species distribution and antifungal susceptibility of Candida isolates during 2002-2008. Of 177 Candida isolates from blood, species distribution was 90 (51%) Candida albicans, 30 (17%) C. parapsilosis, 22 (12%) C. glabrata, 6 (3%) C. tropicalis and 29 (16%) other Candida spp.. Of 162 Candida isolates from vascular catheter, species distribution was 87 (54%) C. albicans, 14 (9%) C. parapsilosis, 36 (22%) C. glabrata, 5 (3%), C. tropicalis, 2 (1%) C. krusei and 18 (11%) other Candida spp.. Of 1889 Candida isolates from urine, species distribution was 1165 (62%) C. albicans, 22 (1%) C. parapsilosis, 484 (26%) C. glabrata, 83 (4%) C. tropicalis, 26 (1%) C. krusei and 109 (6%) other Candida spp.. Of 782 Candida isolates from stool, species distribution was 425 (54%) C. albicans, 3 (1%) C. parapsilosis, 103 (13%) C. glabrata, 28 (4%) C. tropicalis, 5 (1%), C. krusei and 218 (28%) other Candida spp.. Both C. albicans and non-Candida spp. isolated from urine increased slightly over the past 7 years. Flucytosine, fluconazole, itraconazole and micafungin still have strong activity against Candida isolates.

Pseudomonas aeruginosa has been reported to be a leading cause of nosocomial infections. We evaluated the characteristics using 903 P. aeruginosa strains that were isolated in Fukuoka University Hospital from 2006 to 2008. Clinical specimens of P. aeruginosa were detected 47.8% from respiratory tract, 24.4% from urine, and were detected from the pus by 18.9%. Age distribution of the patients was 70 years old or older. Antimicrobial susceptibility ratio of isolates of P. aeruginosa from the outpatients against all agents tested except amikacin had more susceptible than it of isolates from the inpatients. In P. aeruginosa isolates detected from the inpatients, susceptibilities to other antibiotics were about 60% though 93.3% and 83.2% were susceptible to amikacin and piperacillin/tazobactam respectively. Isolation rate of multidrug-resistant P. aeruginosa (MDRP) in all clinical isolates was 3.3%. The report of carbapenem use was obligated in our hospital from the experience of the outbreak by MDRP in 2007, and the infection control team came to regulate the administering of antimicrobial agents more than before. Afterwards, the amounts of the antimicrobial agent use except piperacillin/tazobactam have decreased to 69% compared with the previous year. The susceptibility rates of various antimicrobial agents except amikacin have recovered by 10-17%. These results suggest that there is a necessity for attending to the drug susceptibility and the amount of the antimicrobial agent use.

Aim: Purple urine bag syndrome is a condition in which the urinary catheter bag turns purple. A tryptophan-indigo hypothesis has been proposed as the mechanism of PUBS, in which bacterial decomposition of tryptophan in gut associated with chronic constipation, bacterial overgrowth in the urinary tract and alkaline urine causes production of indigo and discoloration. We considered that further investigation of cases was needed., Methods: We investigated 6 cases exhibiting PUBS (3 males and 3 females)., Results: All cases had chronic constipation. Oral ingestion was impossible in one case. PUBS disappeared after antibiotic treatment (3 cases) or spontaneously (one case). Alkaline urine and indicanuria were not found in all cases that showed the disappearance of PUBS. In bacterial culture of urine during the exhibition of PUBS, Enterococcus faecalis was isolated together with Morganella morganii (3 cases) and Pseudomonas aeruginosa (one case). Single infections by Klebsiella pneumoniae or Citrobacter species were also found. After disappearance of PUBS, infected bacterial species changed but no cases showed sterile urine. Urine and blood alpha-amino-n-butyric acid levels reduced after the disappearance of PUBS whereas tryptophan levels did not show related changes. In one case, blood protein concentration increased after the spontaneous disappearance of PUBS. Indicanuria and alkalization of urine from urinary catheter bag were more intense than of fresh urine., Conclusions: The present results generally support the 'Tryptophan-indigo hypothesis'. Furthermore, it was suggested that additional factors associated with the occurrence of PUBS are an environment that facilitates specific bacterial growth in a hospital as well as abnormal metabolism relating to alpha-amino-n-butyric acid and reduced protein synthesis in patients.

Urine samples collected from 422 males and 53 females visiting a clinic in Kawasaki City who were suspected to have sexually transmitted infection were tested for the detection of Chlamydia trachomatis and Neisseria gonorrhoeae by BD ProbeTecET (SDA method). The detection rates of C. trachomatis by the SDA method and polymerase chain reaction (PCR) method (control) were as high as 98.1% for C. trachomatis, and as high as 99.4% for N. gonorrhoeae, and the concordance rate of detection of both bacterial species was high. The detection sensitivity and specificity of the SDA method were 90.6 and 99.3%, respectively for C. trachomatis and 98.7% and 100% for N. gonorrhoeae, when PCR was used as the standard method. There were no differences in these results between males and females. The number of patients showing a discrepancy of the results obtained between the SDA method and the PCR method was 9 for C. trachomatis and 1 for N. gonorrhoeae, but the results of redetermination by the SDA method tended to coincide with those of the PCR method. Urine samples tested by the SDA method were positive for N. gonorrhoeae even in patients in whom the culture of secretions from the male urethra was negative for N. gonorrhoeae. Based on these results, the BD ProbeTecET (SDA method) was confirmed to have the equivalent capability to the PCR method for the detection of C. trachomatis and N. gonorrhoeae in urine samples.

Aim: Patients in nursing homes are becoming more and more elderly and also prone to infectious diseases. It is important to select proper antimicrobial agents in treating such patients because of the increase in drug-resistant bacteria in recent years., Methods: Pathogenic aerobic bacteria were isolated from cultures of the pharyngeal swab obtained from patients with acute febrile episodes and those with chronic febrile conditions with a repetition of fever or continuing low-grade fever. Isolation of pathogens was also carried out in patients with a urinary tract infection that was resistant to treatment. Isolated bacteria were tested for sensitivity to commonly used antimicrobial agents., Results: Pathogenic bacteria were isolated from 33% of the patients with acute febrile episodes, while they were isolated from 75% of the patients with chronic febrile conditions. The number of major pathogenic bacteria from 85 isolates were methicillin-resistant staphylococcus aureus (MRSA) 13, methicillin-sensitive staphylococcus aureus (MSSA) 6, Streptococcus pneumoniae 8, beta-hemolytic streptococcus 5, Klebsiella pneumoniae 10, and Enterobacter cloacae 6. Only two isolates of Streptococcus pneumoniae were penicillin-sensitive (PSSP), while the others were penicillin-resistant (PRSP) (1) or penicillin-insensitive (PISP) (5). Among these 8 isolates, 5 were resistant to levofloxacin (LVFX). Escherichia coli was isolated from the pharyngeal swab of 2 patients, one of the 2 isolates being resistant to LVFX. Escherichia coli was isolated from 5 patients with urinary tract infection and 5 of the 6 isolates were resistant to LVFX; with one of them being extended spectrum beta-lactamases (ESBL)., Conclusion: The frequency of isolation of antimicrobial-resistant pathogens was extremely high among elderly patients in our nursing home compared to values reported from a nation-wide survey recently carried out in Japan. In particular, attention should be focused on the resistance of bacteria to fluoroquinolones.

We analyzed Pseudomonas aeruginosa isolates in Gifu prefecture between September and October 2004. We conducted antimicrobial susceptibility test for 266 strains isolated from 8 medical institutes and 1 clinical laboratory, based on broth microdilution method. The MIC50 and MIC90 of piperacillin, amikacin, imipenem, and ciprofloxacin were 4 and 64, 4 and 8, 1 and 16, 0.25 and 8 microg/mL, respectively. The strains isolated from urine had higher MIC level in comparison with from sputum, which was remarkable in penicillins, cephalosporins and fluoroquinolones. We isolated 7 strains of multi-drug resistant Pseudomonas aeruginosa (MDRP), in which 3 strains showed under 16 microg/mL in MIC against anti-MRSA (methicillin-resistant Staphylococcus aureus) drug arbekacin. Continuous surveillance would be needed for antimicrobial resistance on P. aeruginosa in Gifu prefecture.

We tested the drug susceptibility to 8 anti-pseudomonal agents of 97 strains of Pseudomonas aeruginosa isolated from urine between January 1998 and May 2004. The results were as follows. 1. Antimicrobial activity was, in order of superiority to biapenem (BIPM), meropenem (MEPM), ciprofloxacin (CPFX), imipenem (IPM), pazufloxacin (PZFX), amikacin (AMK), ceftazidime (CAZ), piperacillin (PIPC). 2. The resistance rate (intermediate+resistance) to carbapenem drugs was 10.3% for BIPM and MEPM, and 13.4% for IPM. Many of the IPM-resistant strains showed crossover resistance with BIPM and MEPM. 3. The resistance rate (intermediate+resistance) to fluoroquinolone drugs was 23.7% for CPFX and 20.6% for PZFX. 4. One strain showed simultaneous resistance to IPM = 16 microg/mL, CPFX = 4 microg/mL, and AMK = 32 microg/mL, and produced IMP-1 metallo-beta-lactamase. Susceptibility of P. aeruginosa isolated from urine developed resistance to fluoroquinolone drugs. It is important to promote appropriate use of antimicrobial agents and continue to survey emerging resistance in the clinical isolates.

We investigated the isolation circumstances of multiple-drug-resistant Pseudomonas aeruginosa (MDRP) in the UOEH hospital and the bacterial analysis of isolated MDRP. From January to October 2003, MDRP was isolated from 2 patients. During this period, the isolation frequency of MDRP was 0.57% (2/350). Case 1 had 2 MDRP isolates from catheter urine, and case 2 had 5 MDRP isolates from pus. Regarding serotype, 2 isolates from case 1 were B type and the other 16 isolates from case 2 were E type. Pyomelanin was produced by 9 isolates of 16 E type isolates. The same PFGE patterns were observed in 2 isolates from case 1; that is, 9 pyomelanin producers from case 2 and the other 7 isolates from case 2, respectively. Metallo-beta-lactamase was produced by 2 isolates from case 1. bla(IMP) was detected from the 2 isolates by PCR, and the clones from case 1 were quite different from the clones from case 2. Regarding the pyomelanin producing isolates from case 2, although the clones were the same genetically, the MICs of imipenem and meropenem increased from 8 to > 32 microg/ml with the progress of time. In the UOEH hospital, 6 patients with MDRP isolates have been isolated so far, but these 6 patients are not correlated with each other. It is important that we detect and report MDRP as early as possible to prevent nosocomial infection.

We investigated an epidemiological study for fungus isolation in our hospital from 1976 to 2000. For 25 years, the total sample number of fungus examination were 64,296, and after 1988, the total sample number increased suddenly. As a whole, the positive ratio was constantly about 40%. When our hospital opened, the obstetrical and gynecological samples showed 38.8% for fungus examination, but recently, samples of the respiratory organ has increased. Ratio of isolation for yeast, Candida albicans was 53.8%, and another yeasts such as Candida glabrata, Candida tropicalis, Candida parapsilosis were 12.5%, 5.3%, and 3.4%, respectively. Recently, isolation of Candida glabrata showed a tendency to increase. For genus Aspergillus, Aspergillus fumigatus was isolated, 48.1%, and Aspergillus nigar, Aspergillus terreus were isolated, 31.4% and 7.5%, respectively. For dermatophytes, Trichophyton rubrum was isolated, 63.6% indermatophytes, and another dermatophytes were Microsporum canis (17.9%), and Trichophyton mentagrophytes (15.9%), respectively. For dermatophytes, isolation of Microsporum canis showed a tendency to increase. Recently, the plural number of species showed a tendency to increase in the samples. Compared with the number of samples at the beginning in our hospital, the plural number of species in the samples increased about six times.

The results of bacteria isolated from the urine of inactive elderly patients with urinary incontinence in 1982, 87, 93 and 95 were compared and the chronological changes of urinary bacteria were analyzed. The following results were obtained: (1) The most common bacteria isolated from the patients in 1995 were D-group Enterococcus, followed by Providencia rettgeri, E. coli, Pseudomonas aeruginosa and Proteus mirabilis. It is thought that the main cause of the more frequent isolation of Enterococcus is an abuse of cephems and fecal contamination. (2) The most common bacteriaisolated from the patients who were managed with an indwelling urethral catheter were Proteus and Providencia species, and their incidence tended to decrease from 1982 to 1995. (3) Infection with Pseudomonas species tended to increase, as did that Staphylococcus species, especially Methicillin-resistant Staphylococcus aureus and then decreased. Based on these findings, it is important to assess and treat dysuria before inserting an indwelling urethral catheter, and to select an antibacterial agent that is effective in elderly patients with urinary incontinence and limited activity.

To investigate the formation of infection stones by 5 bacterial species isolated from patients with urinary calculi, such as Proteus mirabilis, Staphylococcus saprophyticus, Pseudomonas aeruginosa, Klebsiella pneumoniae and Escherichia coli, a zinc disc was implanted and 10(6) bacteria were surgically inoculated at the same time into the bladder of male Wistar rats (10 rats per group). Urinary pH was measured on days 1, 3, 5 and 7. The rats were killed on day 7 and the bladder examined for the formation of infection stones. Infection stones were formed in all rats inoculated with P. mirabilis (mean stone weight 29.7 mg), and in 60% of those inoculated with S. saprophyticus (9.7 mg). Small bladder stones were formed in 30% of the rats inoculated with K. pneumoniae and 25% of those inoculated with P. aeruginosa. No bladder stones were formed in the rats inoculated with E. coli. Urinary pH was increased by P. mirabilis and S. saprophyticus but was unaffected by the other species. In conclusion, P. mirabilis and S. saprophyticus play a significant role in the formation of infection stones.

The relationship between infection stone and Corynebacterium species was investigated in vitro and in vivo. Urease activity of urease-splitting Corynebacterium species was evaluated by 2 methods; an increase in pH of human urine after inoculation of Corynebacterium species and direct measurement of urease activity of 10(7) CFU organisms from amounts of ammonia by indophenol method. Formation of infection bladder stone was induced in male Wistar rats by implanting a zinc disc and inoculating 10(6) CFU organisms surgically into the bladder. Urine was alkalinized by the inoculation of Corynebacterium renale, C. pilosum and group D2 Corynebacterium. C. renale and C. pilosum had strong urease activity, and group D2 Corynebacterium had moderate activity. C. pseudodiphtheriticum did not produce the elevation of urinary pH and had little urease activity. Infection stones were formed in 100% of rats by inoculation of C. renale and C. pilosum and 88% of rats by group D2. Urinary pH was elevated in all inoculated rats. In conclusion, C. renale, C. pilosum and group D2 Corynebacterium may play a role in formation of infection stones.

To clarify the relationship between glitter-cells and urine osmotic pressure, neutrophils were isolated from venous blood of 4 normal volunteers and incubated in urine specimens with different osmotic pressures. Glitter-cells were detected in the urine with 100-400 mOsm/l. The rate of glitter-cells in neutrophils incubated for 4 hours were significantly less than those incubated for 0 minute (p < 0.05). We clinically investigated the relationship between the presence of glitter-cells in fresh urine and the site of urinary tract infection (UTI) or organisms isolated from urine. Fresh urine samples were obtained from 95 patients (55 male and 40 female). Urine samples were examined for bacteriuria by the quantitative culture method, and significant bacteriuria was defined as more than 10(5)/ml of bacilli for midstream urine or as more than 10(4)/ml for urine collected by catheterization. Leukocytes and glitter-cells in the unspun and unstained urine were counted on a disposable slide using a counting chamber. Patients with upper UTI showed a significantly higher incidence of glitter-cells than patients with lower UTI (p < 0.05). Patients with glitter-cells had a significantly higher incidence of polymicrobial infections than those without glitter-cell (p < 0.05). These findings suggest that patients with glitter-cells in the urine show upper UTI (pyelonephritis) and polymicrobial infections.

Ecological behaviors of uropathogenic bacteria in the infected urine were characterized, and they were compared to those in vitro cultivation. The organisms directly collected from the infected urine revealed no piliation, but when they were isolated and cultured in heart infusion broth (HIB), the pili appeared. The pili were identified as Type I pili as examined by immunogold electron microscopy. Although the organisms directly collected from the infected urine did not have pili and did not agglutinate against anti-Type I pili anti-serum, the whole cell lysate analysis by western blotting revealed that the organisms express Type I pili antigen intracellularly. The organisms in some cases revealed a capsule formation in the urine, but it disappeared when cultured in vitro. Hemolytic Escherichia coli were isolated from 9 cases out of 20 with acute cystitis, however, hemolysin was not detected from the infected urine and the organisms. When they were cultured in HIB, all strains produced the hemolysin. The composition of outer membrane proteins (OMPs) of the organisms in the infected urine was clearly different from those in vitro cultivation. As the most prominent finding, a 70 kDa OMP of the organisms in the infected urine disappeared in vitro. Antibacterial activities of several drugs examined in healthy human urine and in HIB were variable from drug to drug. The mean value of minimum inhibitory concentration (MIC) of ofloxasin was 25 times higher in the urine than in HIB, whereas the MIC of erythromycin was almost equal in both conditions.

This study was conducted to invesitigate the relative frequency of Staphylocossus spp. and coagulase negative Staphylococci (CNS) isolates from urine at Juntendo University Hospital in Tokyo. The frequency of Staphylococci spp. was encountered in about 10% of the cases from 1984 to 1994 (except 1992 and 1993), and no significant annual changes were observed. The frequency of Staphylococcus aureus has been gradually increasing from 1986. The relative frequency of CNS (321 strains) from 1989 to 1994 were as follows; Staphylococcus epidermidis 36%, Staphylococcus saprophyticus 26%, Staphylococcus haemolyticus 22%, Staphylococus caprae 8%, and other CNS 8%. S. saprophyticus, most of which were isolated from female out-patients, were suggested to be the important pathogen of acute urinary tract infections. All of S. caprae were isolated from male patients. Most of S. epidermidis isolated from inpatients of urinary tracts with other bacteria.

This study was conducted to investigate the antimicrobial susceptibilities of the strains of coagulase negative Staphylococci (CNS) isolates from urine at Juntendo University Hospital in Tokyo from 1989 to 1994. The susceptibility testing were performed according to the broth dilution method standerdized by the Japan Society of Chemotherapy. The following bacteria were tested; Staphylococcus epidermidis (59 strains), Staphylococcus haemolyticus (42 strains), Staphylococcus saprophyticus (30 strains). The antimicrobial agents tested wre as follows; Oxacillin, Cefazolin, Imipenem, Flomoxef, Gentanicin, Tobramycin, Arbekacin, Clindamycin, Tetracycline, Minicycline, Vancomycin, Sulfamethoxaxole-Trimethoprim and, Ofloxacin. 1. 100% of S. caprae, 62% of S. haemolyticus and 42% of S. epidermidis were resistant to Oxacillin. All strains of S. saprophyticus were sensitive to Oxacillin. 2. S. saprophyticus showed the highest sensitivities to all anti-microbial agents. 3. All strains of S. caprae were resistant to Tobramycin and Clindamycine. 4. Vancomycin and Arbecacin has strong antimicrobial activities to all CNS. S. saprophyticus, which is the pathogen of acute urinary tract infections, showed high sensitivities to many antimicrobial agents. On the other hand, S. haemolyticus and S. caprae, which are the predominate microorganisms of bacteriuria of inpatients, showed high resistance rates to antimicrobial agents.

Routine urinalysis is performed as a screening test for urinary tract infection (UTI) in out-patients or in-patients. We assessed the usefulness of microscopic examination of unspun and unstained urine using a disposable slide with counting chambers (Kova Slide 10 grid, Miles-Sankyo) for diagnosis of significant bacteriuria. 173 fresh urine samples were obtained from 173 subjects (89 male and 84 (female), including 117 inpatients, aged from 0 to 96 years. Urine samples were examined for bacteriuria by the standard culture method and counting chamber method. Significant bacteriuria was defined as 10(5)/ml or more of bacilli for midstream urine and urine collected by bags and 10(4)/ml or more for urine collected by catheterization and from indwelling catheters. Urine leukocytes were also counted on disposable slide. The rapid dipstick test (N-multistix-SG-10, Miles-Sankyo) of leukocyte esterase activity and nitrite were measured in the urine specimens read by a photometer (Clinitek-10, Miles-Sankyo). Significant bacteriuria was diagnosed by standard culture method in 67 urine samples. Close correlation was obtained between bacterial counts determined by the bacterial culture and counting chamber method (Spearman's correlation coefficient p < 0.001). Sensitivity and negative predictive value for significant bacteriuria were 94.0 and 95.1%, respectively, when bacteriuria or pyuria was present in the counting chamber. Dipstick test had a sensitivity and negative predictive value of 86.6 and 89.9%, respectively, when either leukocyte esterase activity of + or more, or nitrite of + was found. In out-patients, both sensitivity and negative predictive value were as high as 100% in counting chamber method. Thus, we can conclude that urine microscopy on disposable counting chambers is a very sensitive, simple, time-saving and lost-effective method for diagnosis of UTI.

Using clinically isolated 114 strains of Pseudomonas aeruginosa that were collected from April to October 1994, activity of antipseudomonal agents against these organisms was determined using the method of liquid microdilution. In addition, antimicrobial activities of the agents were graded according to serological groups of organisms. The results of this study are summarized as follows. 1. Many strains of P. aeruginosa were isolated mainly from sputum, pus and urine. 2. Serological group G organisms of sputum origin, group I of pus and bile origin, and group E of urine origin were isolated most frequently. 3. The most powerful antipseudomonal agent was cefclidin. Its MIC50 and MIC90 were 0.78 and 6.25 micrograms/ml, respectively. The second most powerful agent was ciprofloxacin whose MIC50 and MIC90 were 0.39 and 12.5 micrograms/ml, respectively. 4. The proportions of resistant strains ranged from 0.9% for cefclidin to 40.4% for ofloxacin. The antipseudomonal agents to which 30% or more of strains were resistant were cefpirome, gentamicin and ofloxacin. 5. Cefclidin showed the most powerful activity against strains that were resistant to ceftazidime, imipenem, gentamicin and ofloxacin. Its MIC90 against all strains resistant to ceftazidime, gentamicin and ofloxacin was 6.25 micrograms/ml. The MIC90 of cefclidin and tobramycin against imipenem-resistant strains was 3.13 micrograms/ml. 6. Group E organisms were found among strains resistant to ceftazidime, gentamicin and ofloxacin at high rates, but no group E strains were found among imipenem-resistant organisms. 7. Agents with highest activities by serological group of organisms were cefclidin against group A, tobramycin and ciprofloxacin against group B, imipenem against group E, ciprofloxacin against group G, and cefclidin and ciprofloxacin against group I.(ABSTRACT TRUNCATED AT 250 WORDS)

We reported a 53-year-old female who was admitted due to partial loss of consciousness. She had been diagnosed as old pulmonary tuberculosis and diabetes mellitus. She was diagnosed as diabetic keto-acidosis on admission. We isolated Escherichia coli in the blood, transtracheal aspiration (TTA) and from the urine. We have experienced 6 cases where the same bacteria was isolated from the blood and TTA at the same time. In all 6 cases, we have found single bacteria in the blood and a few other bacteria in TTA. Blood culture is the most certain method to detect the origin of infectious diseases. But the compromised host, as in this case, has multifocal infections in many cases. In order to understand the pathological aspects of the infection, we must obtain many kinds of samples and as many as possible.

The role of clarithromycin (CAM) in biofilm formation has recently been reported. Inhibition of the production or promotion of the dissolution of the glycocalyx, a major component of biofilm, has been implicated in its mechanism of action. However, the details remain unclear. We used an experimental model of complicated urinary bladder infection and automated simulation of the variations in urinary antimicrobial concentration to study the efficacy of CMA in inhibiting biofilm formation and obtained the following results. 1) Prior to biofilm formation, Pseudomonas aeruginosa (P. aeruginosa) was exposed to ciprofloxacin (CPFX, MIC: 8 micrograms/ml), which was active against the organism, at a dose of 200 mg t.i.d. for 7 days. The bacteria were apparently eradicated from the culture medium in the experimental model of bladder infection (model bladder) after 32 hours. However, when the medium was changed to eliminate the antimicrobial agent on Day 7, bacterial regrowth was initiated after 4 hours. Scanning electron microscopy demonstrated sequential biofilm formation on the surface of glass beads in the model bladder diverticulumn, suggesting inside the biofilm were a source of regrowth. 2) Prior to biofilm formation, P. aeruginosa was also exposed to CAM alone, which has no antimicrobial activity against the organism (MIC: > 128 micrograms/ml) at a dose of 200 mg t.i.d. for 7 days. In this situation, CAM was not active against P. aeruginosa and the bactericidal concentration in the model bladder did not decrease markedly, reaching the initial level (10(7) CFU/ml) within 48 hours. However, although numerous bacteria were attached to the glass beads in the diverticulum, no biofilm was formed. 3) Exposure to a combination of CPFX and CAM (each at 200 mg t.i.d. for 7 days) resulted in the eradication of bacteria from the model bladder at 32 hours, and no bacterial regrowth was demonstrated after the medium was exchanged on Day 7. In addition, no biofilm was formed and the bacteria did not become attached to the glass beads. 4) The content of alginate, a major component of P. aeruginosa biofilm, was measured per 5 glass beads on Day 3, 5, and 7 after starting drug administration. The alginate content increased with time when CPFX was given alone at a dose of 200 mg t.i.d..(ABSTRACT TRUNCATED AT 400 WORDS)

With the view of making an adaptation study for biofilm of the urinary tract, we devised a computer-controlled highly complicated bladder model (biofilm model of the urinary tract) that allowed the simulation of time-course changes in the urinary concentration of antimicrobial agents. Clarithromycin (CAM), which is reported to have an anti-biofilm action, was examined at urinary levels approximating clinical concentrations and its effect on biofilm was determined. The following results were obtained. 1) Ofloxacin (OFLX, 200 mg x 2/day, MIC; 8 micrograms/ml), which is active against Pseudomonas aruginosa, caused apparent microbial elimination from the model at 42 hours, but bacterial regrowth occurred 4 hours after withdrawal of this agent. No disappearance of the biofilm was noted with OFLX suggesting that this was the cause of bacterial regrowth. 2) The combination of OFLX (200 mg x 2/day) and CAM (MIC; > 128 micrograms/ml, 200 mg x 2/day) on anti-biofilm agent, with no effect on P. aeruginosa, eliminated bacteria from the bladder model more rapidly and prolonged the regrowth time to 10 hours after withdrawal of the antimicrobial agents. Disappearance of most of the biofilm and only slight microbial adhesion was noted. 3) The combination of OFLX (200 mg x 2/day) and CAM (400 mg x 3/day) caused microbial elimination from the bladder model with no regrowth at 30 hours after withdrawal of the antimicrobial agents. The biofilm disappeared completely and no microbial adhesion was noted. 4) CAM alone (400 mg x 3/day) allowed microbial recovery to the initial level within 48 hours after withdrawal, but led to disappearance of the biofilm and the adhesion of microbes without a glycocalyx. 5) These results suggest that the anti-biofilm action of CAM is dose-dependent, and that combined use of an appropriate antimicrobial agent and anti-biofilm agent like CAM may be effective for biofilm infections of the urinary tract.

For prevention of group B streptococcal (GBS) infection in neonates, GBS carriers of pregnant women should be detected. Procedures for detection of carriers were needed to get higher positive rate and to be handled with easy. Urine samples were shown to detect higher rates than vaginal smear samples in pregnant women. GBS in the urine were adsorbed to charcoal granula for easier transporting. These adsorbed charcoal were tested as to whether they could be preserved with different temperatures and days. Urine with 10(1) CFU/ml of GBS added could be detected after 7 days at temperatures of 4, 25, 37, 42 degrees C. The cultures of this procedure with the cultures of urine sediment of pregnant women were compared. Detecting rates of this procedure and urine sediment were 177 (26.2%) and 136 (20.1%) of 676 pregnant women, respectively. These data suggested that adsorbed charcoal culture was useful with higher positive rates for detection of the GBS carriers in pregnant women.

It is known that the neonatal group B streptococcal disease in most cases is acquired by contamination during passage through the birth canal at the time of delivery. Therefore, to investigate the influence of colonization of group B streptococci in the female genital tract by age, we carried out the isolation of group B streptococci from the urine of healthy children of school age group, and those age groups were as follow: kindergarten primary school students, junior high school students, high school students and women over twenty years of age. The isolation rates of group B streptococci from the urine of the healthy persons by each group were as follow: 12.5-18.2% in 93 females and 0-0.3% in 90 males in kindergarten aged 3-5 years, 6.4-25.3% in 285 females and 0.3-3.2% males in primary school aged 6-11 years, 11.8-19.1% in 183 females and 0-7.1% in 233 males in junior high school aged 12-14 years, 16.4-23.7% in 416 females and 2.6-7.1% in 114 males in high school aged 15-17 years, and 35.0-50.0% in adult females aged 20-60 years. Isolation rates according to sex were 2.2% in 756 males to 17.5% in 977 females of aged 3-17 years, and there was a clearly highly significant difference between males and females (p < 0.01). Group B streptococci isolated from urine of 1,818 persons were 220 strains (12.1%), and the 146 (66.4%) in all strains could be typed by the serological test.(ABSTRACT TRUNCATED AT 250 WORDS)

This study was conducted to investigate susceptibilities of urinary tract isolates to antimicrobial agents examined at 123 hospital laboratories in Japan from September to December in 1991. In this study, identification and susceptibility testing were carried out at each laboratory. The susceptibility testing was performed according to the disk diffusion method recommended by National Committee for Clinical Laboratory Standards. MRSA showed high resistance rates to all agents tested except to minocycline (MINO). MSSA had good susceptibility to imipenem (IPM) and cephalosporins with susceptibility rates higher than 90%. Enterococcus faecalis was susceptible to IPM, ampicillin and piperacillin (PIPC) and highly resistant to cephalosporins, gentamicin (GM), erythromycin, clindamycin and MINO. Escherichia coli, Klebsiella pneumoniae and Proteus mirabilis were susceptible to beta-lactam drugs, GM and ofloxacin (OFLX). High susceptibility rates were observed for strains of Enterobacter cloacae, Citrobacter freundii to IPM, GM, and OFLX and of Serratia marcescens for IPM and GM. Pseudomonas aeruginosa strains were highly susceptible to IPM, ceftazidime, AMK, cefsulodin, cefoperazone, PIPC in this order.

Using a severely complicated in vitro bladder model with a computer-controlled autosimulation system for antimicrobial concentration in urine, we examined the significance of the peak concentration value and time above MIC, and obtained the following results. 1. Regarding the initial bactericidal speed, the higher the peak concentration value was, the more quickly the bacterial concentration decreased. 2. Concerning the minimum value of bacterial concentration in the model bladder, the higher the peak concentration value was, the more the level tended to decrease. 3. Regarding inhibition of regrowth of bacteria (ERT) 1) When a particular antimicrobial agent had no PAE on bacteria, the viable bacterial count curve was similar in cases of peak antimicrobial concentration values of higher than 8 MIC. 2) On the other hand, when an antimicrobial agent had PAE on bacteria, regrowth of bacteria occurred quickly below a certain peak concentration (24 MIC or 64 MIC value), although there was some difference depending on the type of antimicrobial agents. Thus, it was suggested that the peak concentration above that level be attained and maintration above that level be attained and maintained for some time were in order to obtain PAE adequately. 4. It is concluded from the aforementioned facts that two factors, a certain high concentration and a certain continued time are necessary for both antimicrobial agent with metabolic disorder and antimicrobial agent with cell wall synthetic inhibition to obtain proper antimicrobial effect. The condition of their necessity differed slightly according to the two different agents.

Mechanisms for purple discoloration of the plastic urine bag in purple urine bag syndrome (PUBS) were investigated. Activities of bacterial indoxyl sulfatase catalyzing the conversion of indoxyl sulfate to indigo (or indirubin) were detected in strong alkaline liquid media but not in normal ones. These enzyme activities were particularly high in simple and combined cultures of Proteus mirabilis and/or Klebsiella pneumoniae. These results suggest that occurrence of PUBS is associated with strong alkaline urine as well as urinary tract infections induced by some species of bacteria with indoxyl sulfatase.

Purple urine bag syndrome is a rare phenomenon in which bags turns purple or blue following catheterization. This phenomenon was reported to occur by Providencia stuartii in the presence of indicanuria. We reported herein three females above 75 years of age having a similar phenomenon. All were bedridden chronically and constipated. In these cases, no growth of P. stuartii was found on repeated urine culture, but Escherichia coli and Morganella morganii were concomitantly isolated during the blue bags. It is suggested that purple urine bag syndrome develops in a combination of E. coli and M. morganii besides P. stuartii.

Anti staphylococcal activity by Pseudomonas aeruginosa was investigated through the use of the reversed agar plate and the filter paper stamp methods. Investigation was also conducted on the dye production of different clinical isolates of Pseudomonas aeruginosa, the relationship between drug susceptibility and anti staphylococcal activity, and the influence of erythromycin on anti staphylococcal activity. Seventy four strains of Pseudomonas aeruginosa were prepared which included 20 strains from pus, 34 strains from sputum and 20 strains from urine. These were then inoculated with methicillin resistant Staphylococcus aureus (MRSA). They were then cultured for 48 hours by using the reversed agar plate and the filter paper stamp methods. Anti staphylococcal activity was observed in 16 strains from pus (80%), 19 strains from sputum (55.9%) and 8 strains from urine (40%). The Pseudomonas aeruginosa strains which have no pigment tended to show poor anti staphylococcal activity. Drug susceptibility was tested using PIPC, AMK, IPM, CFS and OFLX. The strains which showed resistance to OFLX tended to show poor anti staphylococcal activity. Erythromycin inhibited the dye production of Pseudomonas aeruginosa but exhibited no effect on anti staphylococcal activity. Consequently, these results suggest erythromycin has exhibited a previously unknown pharmacological effect, furthermore, anti staphylococcal activity was not caused by pigmentation only.

We have reported a method for detection of Chlamydia trachomatis by polymerase chain reaction (PCR) with two oligonucleotides based on sequences within the major outer membrane protein gene from C. trachomatis serovar L2. In the previous report, in addition to treatment of the mixture of first-voided urine (FVU) sediment and 1 ml of urine with proteinase K. DNA purification by phenol extraction was necessary for preparation of template DNA for PCR. In this study, FVU sediment was suspended in 1 ml of Chlamydiazyme dilution buffer and a part of the suspension was treated with proteinase K for DNA extraction. The DNA extraction solution could be used as template for PCR without purification of DNA by phenol extraction. One hundred FVU specimens obtained from male urethritis patients were examined with the two methods (PCR and IDEIA) for detection of C. trachomatis. In 33 of 100 specimens, the DNA fragments of C. trachomatis was amplified by the PCR and in 32 of 100, the chlamydial antigen was detected by IDEIA. The positive and negative coincidence rate of the PCR to IDEIA were 93.8% (30.32) and 95.6% (65/68) respectively, resulting in a high overall coincidence rate at 95%. Thus, the improved method with PCR using FVU as a specimen is proved to be a useful, non-invasive diagnostic tool for diagnosis of chlamydial urethritis.

The effects of chlorhexidine digluconate preadded to urine samples for a neuroblastoma-screening test in preventing the break-down of creatinine (Cre), vanillylmandelic acid (VMA), and homovanillic acid (HVA) by creatinine-cleaving bacteria and the influence of the added disinfectant on the HPLC determination of urinary Cre, VMA, vanillillactic acid (VLA) and HVA. Laboratory or field experiments showed that preadded disinfectant (0.02% volume) nearly completely inhibited the growth of the bacteria and satisfactorily protected urinary Cre, VMA, and HVA from bacterial decomposition. Chlorhexidine digluconate was comparable to benzalkonium chloride in its inhibitory effects on the growth and activities of creatinine-cleaving bacteria. Unlike benzalkonium chloride, chlorhexidine digluconate added to urine samples gave no troubles in subsequent analytical processes. Addition of the disinfectant (0.02% volume) to standard Cre, VMA, VLA, and HVA solution did not affect retention times and sensitivities in HPLC analyses. To estimate influences of the added disinfectant on the serial HPLC determinations of VMA, VLA, and HVA on a large number of urine samples (40, 80, 120, 160, 200, or 5, 500), HPLC analyses were performed on standard VMA, VLA, and HVA solution with repeated injection of the disinfectant in great amounts into a column. The results showed no appreciable changes in the retention times and sensitivities of VMA, VLA, and HVA, and almost complete elution of the disinfectant retained on the column were possible with water-methanol (1:1, 2:8) or methanol washing.(ABSTRACT TRUNCATED AT 250 WORDS)

Neisseria gonorrhoeae was detected from first-voided urine sediments of male patients with urethritis by polymerase chain reaction (PCR). Urine and urinary sediment were treated with proteinase K, and DNA was further purified by phenol extraction. Two oligonucleotides based on sequences within a ribosomal RNA gene from N. gonorrhoeae were used as primers for the PCR. A DNA fragment of 206 bp specific for N. gonorrhoeae was amplified by PCR and detected by agarose gel electrophoresis. In 19 specimens of urine sediments collected from 21 patients in whom N. gonorrhoeae was isolated from urethral swab by culture, 206 bp DNA fragment was amplified by PCR. In all specimens of urine sediments from 24 patients in whom cultures for N. gonorrhoeae were negative, no DNA was amplified by the PCR. The overall coincidence rate between the PCR for detecting N. gonorrhoeae in first-voided urine sediments and culture in urethral swab was 95.6% (43/45). PCR procedure for detection of pathogens from first-voided urine sediments would be noninvasive and would be applied for the diagnosis of gonococcal urethritis and chlamydial urethritis.

The effectiveness of oral administration of an antibacterial alone was compared with that of intravenous administration of an antibiotic in patients with benign prostatic hyperplasia undergoing transurethral resection of the prostate (TUR-P). In group A (23 patients), surgery was carried out by administering 200 mg of ofloxacin (OFLX) alone, during the day before surgery and in the morning of the day of surgery and three times a day from the morning after surgery for two days, making the total dose 1,800 mg, and total period of administration 3 days. In group B (22 patients), 1 g of Cefotetan (CTT) was intravenously infused twice a day for 3 days from the operative day, making the total dose 6 g. The patients in both groups were given no other antibacterials or antibiotics. The maximum body temperature was recorded for post-operative 2 weeks. Negativization of bacteria by bacterial culture in the urine and disappearance of the pyria, were followed up on an outpatient bases. The mean maximum body temperature up to post-operative week 2 was 37.1 +/- 0.3 degrees C and 37.0 +/- 0.3 degrees C in groups A and B, respectively. The mean number of days required until negativization of bacteria in the urine was 41.5 +/- 38.9 days and 43.3 +/- 24.9 days in groups A and B, respectively. The mean number of days required until disappearance of the pyuria was 76.5 +/- 23.0 days and 68.2 +/- 17.9 days in groups A and B, respectively. No significant differences were noted by Student's t-test between groups A and B.(ABSTRACT TRUNCATED AT 250 WORDS)

Chlamydia trachomatis was detected from first-voided urine sediments of 97 male patients with urethritis by polymerase chain reaction (PCR). Since urine and urinary sediments only treated with proteinase K inhibited DNA amplification by PCR, DNA was further purified by phenol extraction and concentrated. Two oligonucleotides based on sequences within the major outer membrane gene from C. trachomatis serovar L2 were used as primers. A DNA fragment of 242 bp specific for C. trachomatis was amplified by PCR and detected by agarose gel electrophoresis. The DNA fragment was amplified by PCR in all specimens of urine sediments from 50 patients with Chlamydiazyme-positive urethral swab. In 38 specimens of urine sediments from 47 patients with Chlamydiazyme-negative urethral swab, PCR was negative. The overall coincidence rate between the PCR for detecting C. trachomatis in first-voided urine sediments and Clamydiazyme in urethral swab was 90.7% (88/97). Detection of C. trachomatis from first-voided urine sediments by PCR was considered to be noninvasive and useful for the diagnosis of male urethritis due to C. trachomatis.

The effectiveness of a single 300-mg dose of tosufloxacin and of 3-day treatment with tosufloxacin (300 mg daily) were compared in a prospective trial in order to clarify the cost-effectiveness of tosufloxacin for acute uncomplicated cystitis. Fifty female patients (25 patients of each group) with acute uncomplicated cystitis received one of these treatment regimens, and the clinical data and follow-up culture results were analyzed. The effectiveness rates of single-dose treatment and 3-day treatment after 3 days were 100% and 96%, respectively. The recurrence rates of single-dose treatment and 3-day treatment after 14 or 28 days were 0% (0/13 patients) and 15% (2/13 patients), respectively. There was no significant difference in the effectiveness rate and the recurrence rate between these two groups, statistically. All isolated strains were eliminated in both regimens. From these results, it is suggested that single-dose treatment of tosufloxacin is effective enough for acute uncomplicated cystitis.

Human cytomegalovirus (CMV) in this study, a polymerase chain reaction (PCR) assay was developed to detect human cytomegalovirus (CMV) from urine specimens taken from renal transplant patients and one congenital CMV infection. Twenty eight urine specimens of 20 renal transplant patients were analyzed by the PCR. CMV infection was detected in 11 specimens (8 recipients). The incidence (40.0%) of PCR positive in the renal transplant patients was highest among the other conventional methods including fluorescent antibody study, anti-IgM EIA and complement fixation test. In two cases, CMV infection could be identified by PCR before antibody development. The PCR procedure is more sensitive and rapid than conventional methods for the diagnosis of CMV. Rapid diagnosis of CMV infection using PCR may be useful in clinical diagnosis and have therapeutic value.

Clinical features of urinary fungal infections were analyzed in 152 patients in whom fungi were cultured from urine and mycologically identified between February, 1989 and June, 1990. The average age was 66 years old, and approximately 70% of the patients were 60 years old or older. Systemic and urinary tract underlying diseases were observed in 145 patients (95.4%) and 70 patients (46.1%) had urinary indwelling catheters. Previous use of antimicrobial agents before isolation of urinary fungi was recorded in 103 patients (67.8%), the 2nd and 3rd generation cephems and new quinolones being most frequent. A mixture of fungi and bacteria was observed in 68 patients (44.7%). Gram-positive cocci were isolated from 50% of them. Of 173 strains of fungi, C. glabrata was the most frequent (31.2%), followed by C. albicans (27.8%), C. tropicalis (17.3%) and T. beigelii (15.0%). All strains of C. glabrata, C. albicans, C. tropicalis and T. beigelii were highly sensitive to flucytosine (5-FC), miconazole (MCZ) and amphotericin B (AMPH), except for 3 strains which were resistant to 5-FC. The consecutiveness was confirmed by repeated culture in 59.4% of the patients. Fungi in the urine, however, spontaneously disappeared in the other patients.

Once a catheter has been passed into the bladder without contamination, there are several possible routes of subsequent infection during drainage period, such as: 1. Entry of bacteria alongside the catheter in the urethra. 2. Introduced bacteria adhered easily to the indwelling catheter and drainage system and colonized. 3. The catheter tip is covered rapidly by various nutrient materials which becomes a good culture medium of stuck bacteria, which is a supply source of bacteria into the bladder urine. 4. Although the motile bacilli ascend very slowly through the stagnant tube and no bacteria ascend against a slowly moving column, rapid transport of organisms occurs in the swirling fluid caused by the passage of rising air bubbles. 5. Continuous urethral catheter drainage permits an average residual urine volume of 7.3 ml. 6. The catheter destroys the antibacterial defense mechanisms of the urinary bladder. The reasons why in many cases of intermittent self-catheterization (CID) urine becomes sterile despite non-sterile procedure, are as follows. In addition to that CIC has none of the disadvantages of the indwelling catheter, it improves the vesical defense mechanisms deteriorated by high pressure voiding. The number of bacteria reintroduced during catheterization is relatively small and they can be eradicated by usual scheduled catheterization within 4-5 hours without residual urine. CIC must be started before trabeculation or diverticuli are formed, in which bacteria remain. Actual determination of residual urine volume after catheterization will help to prescribe a rational program of CIC.