Characteristics of Multidrug-Resistant Corynebacterium spp. Isolated from Blood Cultures of Hospitalized Patients in Japan.

Corynebacterium is a genus consisting of Gram-positive, rod-shaped bacteria, that is wildly distributed in nature. We report the epidemiological characterization of Corynebacterium spp. isolated from blood specimens at the Kurume University Hospital, between June 2008 and November 2011. Twenty-two strains that were likely Corynebacterium spp. were isolated from 22 hospitalized patients, of which 12 (54.5%) were identified as Corynebacterium striatum. Minimum inhibitory concentration tests were performed after biochemical and genotypic identifications. Biofilm production was detected using a 96-well microplate assay. The dissemination of C. striatum was investigated using pulsed-field gel electrophoresis (PFGE). All strains showed the tendency to be resistant to multiple drugs except vancomycin. Fourteen (82.4%) strains, including 9 C. striatum strains were capable of producing biofilms. Four distinct PFGE patterns were detected among C. striatum strains; 6 of which were identified as dominant pattern A (defined in this study) and had high biofilm production ability. During the 3-year monitoring period, these strains might have repeatedly infected the patients or could have readily colonized the hospital environments. C. striatum appeared to be a potential risk factor for bloodstream infections in hospitalized patients. More surveillance and enhanced control strategies are necessary to decrease Corynebacterium spp. infections in hospitals.

Infection control for a methicillin-resistant Staphylococcus aureus outbreak in an advanced emergency medical service center, as monitored by molecular analysis.

A methicillin-resistant Staphylococcus aureus (MRSA) outbreak occurred in an advanced emergency medical service center between 2010 and 2011. Our objective was to evaluate the status of the MRSA outbreak, as monitored by molecular analysis. Twenty-eight MRSA strains were isolated from blood samples from 11 patients, from other specimens (pharynx, nasal cavity, etc.) from 12 patients, from two environmental samples, and from the skin, middle nasal meatus, and urine of one patient each from other wards. Pulsed-field gel electrophoresis (PFGE) was performed to evaluate horizontal transmission. Molecular typing by PFGE showed that the 28 MRSA strains presented 7 patterns in total, and that 11 of the MRSA strains had the same PGFE pattern. Unselective use of intranasal mupirocin ointment, MRSA monitoring for new inpatients, and prevention of direct or indirect contact infection were performed. However, the number of inpatients with MRSA did not quickly decrease, and additional molecular typing by PFGE showed that 10 of 19 MRSA strains found (5 of 6 from blood, 5 of 13 from other specimens) were the same as those found previously. Lectures and ward rounds were performed repeatedly, and staff participation in ward rounds was suggested. Finally, the number of inpatients with MRSA significantly decreased more than 6 months after the intervention. Although the MRSA outbreak was thought to have ended, follow-up molecular typing by PFGE showed that horizontal transmission persisted. Our data suggest that various combinations of infection control measures are essential when dealing with an MRSA outbreak, and monitoring by molecular analysis using PFGE is useful to identify the status of the outbreak.

[A case with Abiotrophia defectiva isolated from postoperative blood culture of a patient with diffuse peritonitis].

We have recently detected Abiotrophia defectiva (A. defectiva) from postoperative blood culture of a patient with diffuse peritonitis. Pleomorphic Gram positive rod appearance and no growth on a standard sheep blood agar led us to suspect aerotolerant anaerobe. From the test results of simplified identification kit for anaerobes, we tentatively reported Clostridium tertium (C. tertium) to the clinician. However, the analysis of 16SrRNA sequence proved it to be A. defectiva. When Gram positive coccus or pleomorphic Gram positive rod are observed on blood culture, it is recommended to incubate Brucella HK (RS) blood agar plates for 24 hours at 35 degrees C both aerobically and anaerobically. The growth both aerobically and anaerobically suggests A. defectiva and Satellitism test and Rapid ID 32 STREP(SYSMEX) testing should be performed to identify the responsible bacteria. A. defectiva is Gram positive streptococcus and requires pyridoxal hydrochloride or L-cysteine to grow. It could be unidentified or misidentified and its particularity often leads incomplete report to the clinician. Our study shows that more careful examination will increase the detection of A. defectiva.

Candidiasis and other oral mucosal lesions during and after interferon therapy for HCV-related chronic liver diseases.

BACKGROUND: Oral lichen planus (OLP) is seen frequently in patients with hepatitis C virus (HCV) infection. The aim of this study was to evaluate the occurrence of oral candidiasis, other mucosal lesions, and xerostomia during interferon (IFN) therapy for HCV infection. METHODS: Of 124 patients with HCV-infected liver diseases treated with IFN therapy in our hospital, 14 (mean age 56.00 ± 12.94 years) who attended to receive administration of IFN once a week were identified and examined for Candida infection and other oral lesions and for the measurement of salivary flow. Serological assays also were carried out. RESULTS: Cultures of Candida from the tongue surfaces were positive in 7 (50.0%) of the 14 patients with HCV infection at least once during IFN therapy. C. albicans was the most common species isolated. The incidence of Candida during treatment with IFN did not increase above that before treatment. Additional oral mucosal lesions were observed in 50.0% (7/14) of patients: OLP in three (21.4%), angular cheilitis in three (21.4%) and recurrent aphthous stomatitis in one (7.1%). OLP occurred in one patient before treatment with IFN, in one during treatment and in one at the end of treatment. 85.7% of the oral lesions were treated with topical steroids. We compared the characteristics of the 7 patients in whom Candida was detected at least once during IFN therapy (group 1) and the 7 patients in whom Candida was not detected during IFN therapy (group 2). The prevalence of oral mucosal lesions (P=0.0075) and incidence of external use of steroids (P=0.0308) in group 1 were significantly higher than in group 2. The average body weight of group 1 decreased significantly compared to group 2 (P=0.0088). Salivary flow decreased in all subjects throughout the course of IFN treatment and returned at 6th months after the end of treatment. In group 1, the level of albumin at the beginning of the 6th month of IFN administration was lower than in group 2 (P=0.0550). According to multivariate analysis, one factor, the presence of oral mucosal lesions, was associated with the detection of Candida. The adjusted odds ratio for the factor was 36.00 (95% confidence interval 2.68-1485.94). CONCLUSION: We should pay more attention to oral candidiasis as well as other oral mucosal lesions, in patients with weight loss during IFN treatment.

[Vibrio vulnificus pollution of imported frozen Black Tiger shrimps in Japan].

The Ariake Sea area of Japan is endemic for Vibrio vulnificus infection. V vulnificus was isolated from slime from tidal flats, seawater, and fish Sea year-round as we reported previously. To identify new routes and factors of V vulnificus infection, we studied V. vulnificus pollution of imported frozen Black Tiger shrimps purchased from a fish market in Kurume, Fukuoka, Japan. V. vulnificus was isolated from 9 of 100 tails (9%) of Philippines products, 3 of 100 tails (3%) of Indonesia products, and 0 out of 100 tails (0%) of Madagascar products. Cytotoxin-hemolysin genes were identified in 7 V. vulnificus strains isolated from patients with V vulnificus septicemia, 9 strains from Philippine products, and 3 strains from Indonesian products. These results suggest that imported frozen Black Tiger shrimps are a new sources of V. vulnificus infection.

Determination of single nucleotide polymorphisms in N-acetyltransferase2 gene using an electrochemical DNA chip and an automated DNA detection system.

An electrochemical DNA chip using an electrochemically active intercalator and DNA probe immobilized on a gold electrode has been developed for genetic analysis. In this study, N-acetyltransferase2 (NAT2) gene polymorphisms (C481T G590A G857A) were determined by the electrochemical DNA chip and the automated DNA detection system that performs hybridization reaction, washing, detection, and data analysis. Human genomic DNAs were extracted from blood and DNA fragments containing the three polymorphisms were amplified by the polymerase chain reaction (PCR) method. Double-stranded PCR products were treated with T7 exonuclease and single-stranded target DNAs were obtained. A sample containing the single-stranded target DNAs was injected into a cassette including the electrochemical DNA chip and set in an automated system. The turnaround time for genotyping with this system was 90 min. A total of 38 samples were automatically genotyped by an SNP determination algorithm. The results of genotype were completely consistent with those determined by the polymerase chain reaction restriction fragment length polymorphism (PCR-RFLP) analysis. Consequently, this method requires no labeling step and has the advantage of realizing a compact and automatic system, and so the system is expected to contribute to personalized medicine based on genotype.

[Antifungal susceptibility of Candida species causing candidemia in Kurume University Hospital and Shaiseikai Hutukaichi Hospital].

Prevention of candidemia has been difficult and empirical therapy may eventually reduce morbidity and mortality. Successful empirical therapies depend on understanding of fungal features and antifungal agents. Susceptibility to amphotericin B (AMPH-B), flucytosine (5-FC), fluconazole (FLCZ), itraconazole (ITCZ), miconazole (MCZ), and micafungin (MCFG) of 41 Candida species isolated from blood were determined. Candida albicans was the most common species (23 species), followed by C. parapsilosis (5 species), C. tropicalis (4 species), C. glabrata (3 species), C. guilliermondii (2 species), C. krusei (1 specie), and Candida spp (3 species). The isolation rates of the drug-resistant (DR) fungi were 5% for 5-FC. The rates of DR and susceptible dose dependent (S-DD) fungi were 0% and 2% for FLCZ, respectively. The rates of DR and S-DD fungi were 0% and 17% for ITCZ, respectively. No shift to resistant species in C. albicans occurred in our hospitals. All C. albicans were susceptible for the antifungal agents examined.

[Seasonal change of Vibrio vulnificus in the slime of tidal flats, seawater, and fishes collected along Ariake Sea, Japan].

To investigate the seasonal change of Vibrio vulnificus in Ariake Sea, Japan, we attempted to isolate V. vulnificus from the slime of tidal flats, seawater, and fishes obtained from three harbors along Ariake Sea. The sample were collected twice a month from January to December, 2001. Also, we determined the biological characteristics of the individual isolates. V. vulnificus were isolated throughout the year, but the isolation ratios were higher during the warmer season from June to October. The isolates in the warmer season were able to grow in culture media containing 0.5% NaCl, whereas those isolated in the remaining seasons could not. Moreover, the isolates in the warmer season showed a greater hemolytic activity than those isolated in the remaining seasons. These results suggest that V. vulnificus isolated in warmer seasons are more vigorous in nature than those isolated in the remaining seasons.

Intravenous ulinastatin therapy for Stevens-Johnson syndrome and toxic epidermal necrolysis in pediatric patients. Three case reports.

BACKGROUND: More effective therapy is needed for the treatment of Stevens-Johnson syndrome (SJS) and toxic epidermal necrolysis (TEN). The clinical efficacy of intravenous ulinastatin therapy was investigated in 3 Japanese pediatric patients with SJS or TEN. METHODS: Ulinastatin was given to 1 pediatric SJS patient and 2 pediatric TEN patients within 7 days (patient 1; SJS), 6 days (patient 2; TEN), or 4 days (patient 3; TEN) after the onset of the skin rash. Ulinastatin was administered intravenously at a dose of 7,500 U/kg/day (maximum dose: 300,000 U/day). No corticosteroids were given. After the skin lesions resolved, the ulinastatin dose was reduced to between 2,500 and 5,000 U/kg/day as maintenance therapy and then the drug was withdrawn. RESULTS: Erythema, fatigue, and fever improved within 12-36 h of starting the ulinastatin infusion, and the skin lesions resolved completely after 4-7 days of ulinastatin therapy. None of the patients had cutaneous or ocular sequelae. No patient developed secondary infection or relapse and ulinastatin therapy caused no side effects. CONCLUSION: Ulinastatin dramatically reduced the febrile period with no adverse effects and was very safe in this study. Ulinastatin appears to be a useful and effective therapy for controlling SJS and TEN without sequelae.

[Drug sensitivity and beta-lactamase producibility of various types of bacteria clinically isolated during the period from December 1999 to February 2000].

beta-Lactamase activity and drug sensitivity were measured in 744 strains from 8 species of bacteria isolated at medical institutions in Chikugo District of Fukuoka Prefecture during the period from December 1999 to February 2000. Nitrocefin test revealed that beta-lactamase was positive in 48% of S. aureus, 7% of H. influenzae, and 92% of M. catarrhalis, and acidometry revealed that penicillinase/cephalosporinase were positive in 13%/14% of E. coli, 22%/8% of K. pneumoniae, 47%/97% of E. cloacae, 3%/65% of S. marcescens, and 10%/36% of P. aeruginosa. Based on the assessment of the MIC values of various types of antibacterial drugs for beta-lactamase-producing strains, there were 11 strains (1 strain of K. pneumonia, 6 strains of E. cloacae, and 4 strains of P. aeruginosa) of class-B beta-lactamase-producing bacteria out of a total of 496 strains of Enterobacteriaceae and Pseudomonas aeruginosa. The results of PCR analysis suggested that 1 strain of K. pneumonia, 1 strain of E. cloacae, and 4 strains of P. aeruginosa produced metallo-beta-lactamase. There was no strain (E. coli and K. pneumoniae) of ESBL-producing bacteria. BLNAR strains, on the other hand, were found in 9% (9/100) of H. influenzae.