A cluster of Candida parapsilosis displaying fluconazole-trailing in a neonatal intensive care unit successfully contained by multiple infection-control interventions.

Objective: This study aimed to investigate and contain a cluster of invasive candidiasis cases caused by fluconazole-resistant Candida parapsilosis (FRC) in a neonatal intensive care unit. Methods: Active surveillance was initiated. Direct observations of hand-hygiene compliance (HHC) among staff were conducted before and after the implementation of hand-hygiene (HH) education. Thirty-five environmental cultures were obtained. Phylogenetic analysis of FRC was performed using Fourier-transform infrared spectroscopy and microsatellite genotyping. Results: A total of 14 patients (mean birth weight = 860 g, gestational age = 25 weeks) infected with FRC were identified using the fully automated analyzer, including 5 with clinical infection (three with catheter-related bloodstream infection, one with cutaneous infection, and one with fatal peritonitis) and 9 with colonization. The HHC rate in nurses before performing a sterile or aseptic procedure significantly improved after the HH education (P < .05). Sinks near the patients were contaminated with FRC. All FRC strains were confirmed to be susceptible to fluconazole using the CLSI method, and the microdilution procedure indicated a trailing effect. Phylogenetic analysis showed that all the fluconazole-trailing isolates from patients were clustered together and had the same genotype. Sinks were successfully decontaminated using accelerated hydrogen peroxide and drainage pipes were replaced. Ultraviolet-C decontamination was applied in the milk preparation room. No new cases were detected after the education and disinfection interventions. Conclusions: Sinks are an important reservoir of C. parapsilosis. Active surveillance, environmental hygiene, and constant staff education on maintaining a high level of HHC are necessary to limit the spread of C. parapsilosis.

Evaluation of MALDI-TOF mass spectrometry coupled with ClinProTools as a rapid tool for toxin-producing Clostridioides difficile.

INTRODUCTION: The performance of MALDI-TOF MS combined with analysis platform for identification of toxin-producing Clostridiodes difficile is yet to be known. METHODS: Between August 2018 and September 2020, 61 isolates from stool specimens of patients with C. difficile-associated diarrhea were analyzed using the MALDI Biotyper system. A C. difficile toxin-producer detection model was developed using ClinProTools. The model was validated using 28 known strains that differed from the isolates used to develop the model. RESULTS: The sensitivity and specificity of the Genetic Algorithm (GA) model using isolates grown on Brucella with hemin and vitamin K (BHK) agar plates were 91.7% and 44.4%, respectively. When isolates grown on cycloserine-cefoxitin mannitol agar were analyzed by the model, sensitivity and specificity were 6.3% and 100%, respectively. The GA model using BHK medium showed the highest discriminatory performance in detection of toxin-producing C. difficile. However, a discrepancy in detection of toxin-producing C. difficile was observed in the results generated when the model was being developed and when the model was validated which suggests that incubation conditions may have affected the results. CONCLUSION: MALDI-TOF analysis using ClinProTools has a potential to be a cost-effective tool for rapid diagnosis and contribute to antimicrobial stewardship by differentiating toxin-producing C. difficile from non-producers.

Screening for Metallo-Beta-Lactamases Using Non-Carbapenem Agents: Effective Detection of MBL-Producing Enterobacterales and Differentiation of Carbapenem-Resistant Enterobacterales.

Metallo-beta-lactamases (MBLs) are enzymes that break down carbapenem antibiotics, leading to carbapenem-resistant organisms. Carbapenemase-resistant Enterobacterales (CRE) is one of them. Outbreaks of CRE infection can occur in healthcare facilities and lead to increased deaths, illness, and medical costs. This study was conducted to detect MBLs using non-carbapenem agents and exclude MBLs among CRE isolates. A total of 3776 non-duplicate sequential Enterobacterales isolates from a single facility were screened between January 2019 and December 2022 using non-carbapenem agents, ceftazidime and cefoperazone/sulbactam. Positive 153 isolates (4.0%) were further tested using carbapenemase-confirmation tests and verified through polymerase chain reaction (PCR) testing. Fifteen imipenemase (IMP)-type MBL-producing Enterobacterales (0.4%) including one susceptible to carbapenems were identified. Moreover, 160 isolates (4.2%) meeting the criteria for CRE were directly subjected to PCR testing. All fourteen CRE isolates with MBLs identified through PCR testing were found to be the same strains screened using ceftazidime and cefoperazone/sulbactam. Screening using ceftazidime and cefoperazone/sulbactam can effectively detect MBL-producing Enterobacterales strains. This screening method showed comparable results to screening with meropenem, potentially serving as a supplementary approach and contributing to differentiating between MBL- and non-MBL-producing CRE strains. Our findings support these screening methods, particularly in regions where IMP-type MBLs are prevalent.

Rapid Detection of SARS-CoV-2 RNA Using Reverse Transcription Recombinase Polymerase Amplification (RT-RPA) with Lateral Flow for N-Protein Gene and Variant-Specific Deletion-Insertion Mutation in S-Protein Gene.

Rapid molecular testing for severe acute respiratory coronavirus 2 (SARS-CoV-2) variants may contribute to the development of public health measures, particularly in resource-limited areas. Reverse transcription recombinase polymerase amplification using a lateral flow assay (RT-RPA-LF) allows rapid RNA detection without thermal cyclers. In this study, we developed two assays to detect SARS-CoV-2 nucleocapsid (N) gene and Omicron BA.1 spike (S) gene-specific deletion-insertion mutations (del211/ins214). Both tests had a detection limit of 10 copies/µL in vitro and the detection time was approximately 35 min from incubation to detection. The sensitivities of SARS-CoV-2 (N) RT-RPA-LF by viral load categories were 100% for clinical samples with high (>9015.7 copies/µL, cycle quantification (Cq): < 25) and moderate (385.5-9015.7 copies/µL, Cq: 25-29.9) viral load, 83.3% for low (16.5-385.5 copies/µL, Cq: 30-34.9), and 14.3% for very low (<16.5 copies/µL, Cq: 35-40). The sensitivities of the Omicron BA.1 (S) RT-RPA-LF were 94.9%, 78%, 23.8%, and 0%, respectively, and the specificity against non-BA.1 SARS-CoV-2-positive samples was 96%. The assays seemed more sensitive than rapid antigen detection in moderate viral load samples. Although implementation in resource-limited settings requires additional improvements, deletion-insertion mutations were successfully detected by the RT-RPA-LF technique.

Personal protective equipment use by health-care workers in intensive care units during the COVID-19 pandemic in Japan: comparative analysis with the PPE-SAFE survey.

Aim: We investigated personal protective equipment (PPE) use and supply shortage, training, and adverse events among health-care workers (HCWs) in the intensive care unit (ICU) during the coronavirus disease (COVID-19) pandemic in Japan and compared the results with an international survey that used the same methodology. Methods: This Web-based survey was carried out from 14 April to 6 May, 2020, in Japan and included HCWs directly involved in ICU management of COVID-19 patients. A survey invitation was emailed using the Japanese Society of Intensive Care Medicine's mailing list. Results: We analyzed 460 valid responses from among 976 responses. The N95/FFP2 mask (77%) was the most frequently used, although half of our respondents reported reuse of single-use N95/FFP2 masks. The median duration (1 h) of uninterrupted PPE use per shift was less than that in the international study. The most common PPE-related adverse event was experiencing intense heat (75%). Logistic regression analysis revealed that being a nurse was independently associated with experiencing intense heat. Conclusion: Shortage of PPE and frequent mask reuse were prevalent during the COVID-19 pandemic in Japan. Intense heat is the most significant symptom, especially for nurses, even with short-duration PPE use. Strategies to protect HCWs from dehydration and intense heatstroke are needed.

Evaluation of presumptive identification of Enterobacterales using CHROMagar Orientation medium and rapid biochemical tests.

BACKGROUND: The use of matrix-assisted laser desorption/ionization time-of-flight mass spectrometry is gradually spreading among large-scale laboratories; however, this method is impractical for small-scale laboratories. In laboratories without access to these rapid identification methods, problems related to them remain unsolved. In this study, we aimed to develop a rapid and inexpensive method to presumptively identify Enterobacterales using CHROMagar Orientation medium. METHODS: The algorithm for presumptive identification of Enterobacteriaceae using CHROMagar Orientation medium was based on our previous studies. Modified property tests for indole, lysine decarboxylase, ornithine decarboxylase, and hydrogen sulfide were performed to evaluate the differentiation of the bacterial species. RESULTS: Using the type strains and clinical isolates, it was possible to conduct the property tests at a low cost, within 4 hours. The spot indole test was performed without any nonspecific reactions for the bacteria forming colored colonies. The presumptive identification of bacteria was thereby possible within 24 hours after specimen submission. CONCLUSION: All these results suggest that the rapid presumptive identification of Enterobacterales is possible with this new identification method using CHROMagar Orientation medium. This is therefore a prompt and economical method that can be used in routine laboratory work.

Moraxella species: infectious microbes identified by use of time-of-flight mass spectrometry.

PURPOSE: To report the clinical manifestations, identification, antimicrobial susceptibilities, and treatment outcomes of ocular infections caused by Moraxella species. STUDY DESIGN: Retrospective study. PATIENTS AND METHODS: The medical records of all patients treated at the Departments of Ophthalmology of the Ogaki Municipal Hospital and the Gifu University Graduate School of Medicine for ocular infections caused by Moraxella species between January 2011 and June 2017 were examined. The stored Moraxella species isolated from ocular samples were identified by matrix-assisted laser desorption/ionization time-of-flight mass spectrometry (MALDI-TOF MS), molecular identification, and the biochemical properties. RESULTS: Sixteen eyes of 16 patients were treated for Moraxella ocular infections. The patients' median age was 72 years. A predisposing systemic or ocular condition was identified in 15 of the patients. Nine of the patients developed keratitis; four, conjunctivitis; and three, blebitis. M lacunata (6 eyes), M catarrhalis (6), M nonliquefaciens (3), and M osloensis (1) were identified by MALDI-TOF MS. All isolates were sensitive to levofloxacin, tobramycin, ceftazidime, ceftriaxone, and cefazolin. Twelve patients with keratitis or blebitis were treated with various topical antimicrobial combinations, and systemic antibiotics were used in 10 of the 12 patients. The mean time for the complete closure of the epithelial defects with keratitis was 24 days. The visual outcomes after treatment were favorable except in 1 keratitis patient who underwent enucleation. CONCLUSIONS: The use of duo-therapy with a combination of fluoroquinolone and cefmenoxime should be considered in cases nonresponsive to monotherapy, such as keratitis and bleb-associated infections. MALDI-TOF MS is useful for the identification of Moraxella to the species level.

Daily Review of Antimicrobial Use Facilitates the Early Optimization of Antimicrobial Therapy and Improves Clinical Outcomes of Patients with Bloodstream Infections.

Insufficient information is available to confirm the beneficial effects of implementing an antimicrobial stewardship program in reducing mortality of patients with bloodstream infections. A single institutional cohort study was conducted to evaluate clinical outcomes after implementation of a daily review of antimicrobials used to treat patients with bloodstream infections. Subjects were allocated to groups receiving either intervention or nonintervention. After implementation of an antimicrobial stewardship program, the day from the onset of infection required to administer effective intravenous antimicrobial treatment was significantly shortened (p=0.022), and the rate of de-escalation was significantly elevated (p<0.001) compared with the nonintervention group. Further, the rate of 30-d death associated with bloodstream infection was siginificantly reduced from 11.4 to 5.4% (p=0.030) compared with the nonintervention group. The incidence of adverse events was significantly lower in the intervention group than in the nonintervention group (7.7 vs. 28.0%, p<0.001). Our present findings suggest that daily review of the use of antimicrobials was highly effective for optimizing early antimicrobial therapy and improved clinical outcomes of patients with bloodstream infections.

Case of late-onset bleb associated endophthalmitis caused by Rothia mucilaginosa.

Rothia mucilaginosa is a gram-positive coagulase-negative coccus of the family Micrococcaceae. Although R. mucilaginosa forms part of the oropharyngeal microflora, it has only recently been isolated in ocular infections. We report a case of a 41-year-old man who developed late-onset bleb-related endophthalmitis (BRE). He had undergone glaucoma surgery 21 years earlier and had a thin-walled cystic bleb prior to the development of endophthalmitis in his right eye. He immediately received intravitreal injections of ceftazidime and vancomycin, topical levofloxacin and cefmenoxime, and intravenous cefozopran. Culture of the aqueous humor specimen identified R. mucilaginosa by 16S rRNA sequence analysis. To our knowledge, this is the first report of late-onset BRE caused by R. mucilaginosa. Our case indicates that R. mucilaginosa can be a cause of late-onset BRE, and that molecular analysis is an accurate method to identify R. mucilaginosa.

Beneficial effects of a cardiac support device on left ventricular remodeling after posterior myocardial infarction: an evaluation by echocardiography, pressure-volume curves and ventricular histology.

PURPOSE: Posterior myocardial infarction (MI) can induce LV remodeling and ischemic mitral regurgitation (IMR). The protective effects of a cardiac support device (CSD) against LV remodeling and IMR after posterior MI have been poorly documented. METHODS: Posterior MI was induced by ligation of the left circumflex coronary artery in beagle dogs. After 7 days, the dogs were randomized to a CSD placement (CSD group, n = 8) or no treatment (CTL group, n = 8). RESULTS: At 3 months after MI, the LV remodeling was less marked and the LV and RV systolic functions were better in the CSD group than in the CTL group. Neither the RV nor LV diastolic function (min dP/dt, Tau and EDPVR) was disturbed by the CSD. IMR was consistently prevented in our canine model. CONCLUSION: Early application of a CSD after posterior MI can attenuate LV remodeling without causing any deterioration of the biventricular diastolic function.

[A retrospective study of the relationship between bacterial numbers from central venous catheter tip cultures and blood cultures for evaluating central line-associated bloodstream infections].

Catheter-related bloodstream infection (CRBSI) is an infectious disease requiring special attention. It is a common cause of nosocomial infections; catheter insertion into the central veins particularly increases the risk of infection (CLA-BSI: central line-associated bloodstream infection). We examined the relationship between the number of bacterial colonies cultured from shredded central venous catheter (CVC) tips and from blood cultures in our hospital from 2011 to 2012. Coagulase-negative staphylococci topped the list of microbe isolated from the CVC tip culture, followed by Pseudomonas aeruginosa, Staphylococcus aureus, and Candida spp. S. aureus and Candida spp., with growth of over 15 colony-forming units in the CVC tip culture, were also detected at high rates in the blood culture. However, gramnegative bacilli (Enterobacteriaceae and P. aeruginosa) did not show a similar increase in colony number in the CVC tip culture. Because microbes adhering to shredded catheter tips are readily detected by culture, this method is useful as a routine diagnostic test. In addition, prompt clinical reporting of the bacterial number of serious CLA-BSI-causing S. aureus and Candida spp. isolated from CVC tips could contribute to earlier CLA-BSI diagnosis.

Moraxella lacunata infection associated with septicemia, endocarditis, and bilateral septic arthritis in a patient undergoing hemodialysis: a case report and review of the literature.

We report the first case of both endocarditis and bilateral septic arthritis in a patient caused by Moraxella lacunata and successful management of the infection with antimicrobial therapy. The route of entry leading to bacteremia may have been the oral cavity given the poor oral hygiene of the patient as evidenced by bleeding gums. We hypothesize that the bacteremia led to septic arthritis and mitral valve infective endocarditis. In this case report, we also review the literature on M. lacunata infections and conclude that this organism should be considered in bilateral septic arthritis in a patient with underlying heart abnormalities and/or with renal failure.

A case of sepsis and meningitis caused by probable travel-related Neisseria meningitidis serogroup B infection: the first report of N. meningitidis ST-4893 in Japan.

Herein we present a case of Neisseria meningitidis-related sepsis and meningitis in a 60-year-old woman. The N. meningitidis strain was identified as serogroup B and sequence type (ST)-4893 by multilocus sequence typing (MLST). The patient in this case had visited France prior to development of symptoms. No meningococcal isolate belonging to ST-4893 has been identified in Japan previously, whereas an ST-4893 strain from France has been reported in the MLST database. These results strongly suggest that this case is likely to have been imported from France.