Molecular Analysis for Potential Hospital-Acquired Infection Caused by Aspergillus Tubingensis Through the Environment.

The identification of Aspergillus species has been performed mainly by morphological classification. In recent years, however, the revelation of the existence of cryptic species has required genetic analysis for accurate identification. The purpose of this study was to investigate five Aspergillus section Nigri strains isolated from a patient and the environment in a university hospital. Species identification by matrix-assisted laser desorption/ionization-time-of-flight mass spectrometry identified all five black Aspergillus strains as Aspergillus niger. However, calmodulin gene sequence analysis revealed that all five strains were cryptic species, four of which, including the clinical strain, were Aspergillus tubingensis. Hospital-acquired infection of the patient with the A. tubingensis strain introduced from the environment was suspected, but sequencing of six genes from four A. tubingensis strains revealed no environmental strain that completely matched the patient strain. The amount of in vitro biofilm formation of the four examples of the A. tubingensis strain was comparable to that of Aspergillus fumigatus. An extracellular matrix was observed by electron microscopy of the biofilm of the clinical strain. This study suggests that various types of biofilm-forming A. tubingensis exist in the hospital environment and that appropriate environmental management is required.

Microbiological analysis concerning the antibacterial effect of atomized Ionless® hypochlorous acid water in a nursery school environment.

INTRODUCTION: In daycare centers, infants come in close contact with each other, and contact, droplet, and mouth-to-mouth infections may occur owing to sharing of toys. Additional effective disinfection methods should be considered aside from wiping with disinfectants-including alcohol or sodium hypochlorite solution-for environmental disinfection of daycare centers. We aimed to examine the usefulness of hypochlorous acid water atomization in the effective disinfection of the classroom environment and toys at a nursery school. METHODS: Environmental cultures of the nursery and toys were prepared to evaluate the species and bacterial load and to assess the contaminated areas. Staphylococcus aureus petri dishes were placed at high-frequency contact sites, and hypochlorous acid water was atomized to achieve a 0.03-ppm atmospheric chlorine concentration. After the atomization, the amount of S. aureus bacteria on the Petri dish and the changes in bacterial count isolated from the environment and toys were evaluated. RESULTS: Hypochlorous acid water atomization was performed for 5 h to avoid condensation. After a 3-h atomization, ≥99.99% of S. aureus was eliminated on petri dishes; furthermore, a significant disinfection effect was observed on environmental bacteria at least 1 h after atomization. For rubber and textile toys, the significant disinfection effect was observed 1 h after atomization, and for plastic toys, the effect was observed 3 h after atomization. CONCLUSIONS: Hypochlorous acid water atomization is a useful strategy to disinfect nursery school classrooms.

Risk Factors and Clinical Characteristics of Patients with Ocular Candidiasis.

Ocular candidiasis is a critical and challenging complication of candidemia. The purpose of this study was to investigate the appropriate timing for ophthalmologic examinations, risk factors for complications of ocular lesions, and their association with mortality. This retrospective cohort study applied, using multiple logistic regression analysis and Cox regression models, to cases of candidemia (age ≥ 18 years) for patients who underwent ophthalmologic consultation. Of the 108 candidemia patients who underwent ophthalmologic examination, 27 (25%) contracted patients had ocular candidiasis, and 7 experienced the more severe condition of endophthalmitis, which included subjective ocular symptoms. In most cases, the initial ophthalmologic examination was performed within one week of the onset of candidiasis with a diagnosis of ocular candidiasis, but in three cases, the findings became apparent only after a second examination within 7−14 days after onset of candidiasis. The independent risk factor extracted for the development of ocular candidiasis was the isolation of C. albicans (OR, 4.85; 95% CI, 1.58−14.90), unremoved CVC (OR, 10.40; 95% CI, 1.74−62.16), and a high ßDG value (>108.2 pg/mL) (HR, 2.83; 95% CI = 1.24−6.27). Continuous ophthalmologic examination is recommended in cases of candidemia with the above risk factors with an initial examination within 7 days of onset and a second examination 7−14 days after onset.

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.

Determination of epidemiology of clinically isolated Cryptococcus neoformans strains in Japan by multilocus sequence typing.

Cryptococcus neoformans and Cryptococcus gattii are the causative agents of cryptococcosis. Despite its importance, our knowledge of the epidemiology of cryptococcosis in Japan remains limited. To establish an epidemiological database on cryptococcosis in Japan, we determined the genetic variability of 44 Japanese clinical isolates of C. neoformans (var. grubii: serotype A) by multilocus sequence typing (MLST). The strains were clinically isolated from 1992 to 2011 in 5 different areas of Japan (the Hokkaido region [n = 1], Kanto region [n = 32], Chubu region [n = 1], Kansai region [n = 1], and Kyushu region [n = 9]). According to the method recommended by the International Society for Human and Animal Mycology cryptococcal genotyping working group, 36 isolates (82%) were identified as sequence type (ST)46. The remaining strains belonged to ST45 (n = 1) and ST47 (n = 1), and 6 isolates belonged to novel independent STs. There was little geographic difference in the ST population. Our present data are still limited; however, because most clinical isolates showed the same MLST profile in Japan, applying the current MLST scheme for Cryptococcus may at times be insufficient for investigating the infection route among outbreak cases. To solve this problem, it may be necessary to investigate other gene loci or develop a novel method with greater discriminatory power. However, in cases in which a strain belongs to a minor ST, our data may serve as useful epidemiological information in Japan.

[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.

[A case of Desulfovibrio desulfuricans cultured from blood in Japan].

We report a case of Desulfovibrio desulfuricans bacteremia in a 60-year-old-man. In our case, anaerobic blood culture bottle turned out positive after five days' incubation. Gram stain showed the presence of slightly-curved Gram negative rod. Suspecting Campylobacter and Helicobacter, we added microaerobic culture while tentatively reporting Campylobacter to the physician. We then added anaerobic culturing with Brucella HK (RS) Agar because microaerobic culture proved the absence of microaerophile. We found small colonies on the third day, then we started anaerobic culture and eventually identified Desulfovibrio desulfuricans. We believe this is the first report of Desulfovibrio desulfuricans cultured from blood in Japan. In case Gram stain shows the presence of spiral bacterium, it is recommended to observe closely considering Desulfovibrio.

Comparison of two control measures of weatherstripping in reducing blowing dust during hospital renovations.

Hospital renovation projects pose risks of invasive infection by fungi from dust that is blown about during the period in question. Control measures to reduce the amount of dust during hospital renovation are thus necessary. Currently, no study has compared different control measures for effectiveness through more than one period of renovation. In this study, we examined the capacities of two control measures of weatherstripping (0.15 mm poly film and adhesive tape) to reduce the amount of blowing dust during two different hospital renovations (in 2008 and 2009). The amount of dust in the air of the hospital before and during the renovation was measured about once a week in both 2008 and 2009, and the between-year and within-year differences were tested. Our study revealed that the weatherstripping used in 2009 (adhesive tape) was significantly more effective than the measures taken in 2008 (0.15 mm poly film) to reduce the amount of dust during the renovations (p < 0.001), while in both years the amount of dust became significantly higher during the renovations than before the renovations. Differences in the effectiveness of weatherstripping during renovations between floors of the hospital were not significant in both 2008 and 2009. The number of Aspergillus-positive samples did not significantly increase compared with the number observed before the start of the hospital renovations (2006-2007) in 2008 and 2009, respectively. The weatherstripping potentially reduced the associated risk of airborne fungal infection.

[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.

[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.