Implementation of MALDI-TOF Mass Spectrometry to Identify Fungi From the Indoor Environment as an Added Value to the Classical Morphology-Based Identification Tool.

Introduction: During the last decades, molds in the indoor environment have raised concern regarding their potential adverse health effects. The genera Aspergillus, Cladosporium, Penicillium, Alternaria, and yeasts, the most common fungi found indoors, include species with high allergenic and toxigenic potentials. Identification of these molds is generally performed by microscopy. This method has, however, some limitations as it requires mycologists with high expertise while identification is often limited to the genus level. Therefore, it is necessary to seek for fast and accurate tools, such as Matrix-assisted laser desorption/ionization time-of-flight mass spectrometry (MALDITOF MS), enabling an identification to the species level and guiding general practitioners in their search for the underlying cause of a health problem. Methods: In this study, 149 fungal air and dust isolates from 43 dwellings in Brussels were taken in collaboration with Brussels Environment RCIB/CRIPI and identified by both microscopy and MALDI-TOF MS in Sciensano's Indoor Mycology laboratory. Spectra obtained via MALDI-TOF MS were compared with data available in an in-house created reference database containing over 1,700 strains from the BCCM/IHEM fungal collection. Results: A total of 149 isolates including 18 yeasts and 131 filamentous fungi were analyzed. Microscopic analysis indicated 18 yeast species and allowed identification of 79 isolates (53%) to the genus level. Only 36 isolates (24%) could be identified to the species complex level. Fifteen molds (10%) could not be identified, and one was indicated as sterile mycelia. No isolate was identified to species level. MALDI-TOF MS analysis identified 137 (92%) of the 149 isolates with a logscore > 1.7. Of these 137 isolates, 129 (87%) were identified to the species level (logscore > 2.0). For only 8 isolates (5%), identification was limited to the genus/section level (1.7 < logscore <2.0), and 12 isolates (8%) could not be identified. Conclusion: A comparison of results obtained with both methods indicates an increased precision in identifications with MALDI-TOF MS analysis for 92% of the isolates, emphasizing its highly added value to the standard microscopic analysis in routine practice. In addition, MALDI-TOF MS also enables to assess the accuracy of microscopic identifications.

Systemic antifungal drug use in Belgium-One of the biggest antifungal consumers in Europe.

BACKGROUND: Reports on the consumption of systemic antifungal drugs on a national level are scarce although of high interest to compare trends and the associated epidemiology in other countries and to assess the need for antifungal stewardship programmes. OBJECTIVES: To estimate patterns of Belgian inpatient and outpatient antifungal use and provide reference data for other countries. METHODS: Consumption records of antifungals were collected in Belgian hospitals between 2003 and 2016. Primary healthcare data were available for the azoles for the period 2010-2016. RESULTS: The majority of the antifungal consumption resulted from prescriptions of fluconazole and itraconazole in the ambulatory care while hospitals were responsible for only 6.4% of the total national consumption and echinocandin use was limited. The annual average antifungal consumption in hospitals decreased significantly by nearly 25% between 2003 and 2016, due to a decrease solely in non-university hospitals. With the exception of specialised burn centres, antifungals are mostly consumed at ICUs and internal medicine wards. A significant decline was also observed in the consumption of azoles in primary health care, attributed to itraconazole. The major part of azoles was prescribed by generalists followed by dermatologists. CONCLUSIONS: In spite of the downward trend in annual use of systemic antifungal drugs, Belgium remains one of the biggest consumers in Europe.

Clonal Spread of Candida glabrata Bloodstream Isolates and Fluconazole Resistance Affected by Prolonged Exposure: a 12-Year Single-Center Study in Belgium.

Candida glabrata is a major cause of candidemia in immunocompromised patients and is characterized by a high-level of fluconazole resistance. In the present study, the acquisition of antifungal resistance and potential clonal spread of C. glabrata were explored at a single center over a 12-year period by analyzing 187 independent clinical C. glabrata bloodstream isolates. One strain was found to be micafungin resistant due to a mutation in the FKS2 gene. Fluconazole resistance remained stable throughout the period and was observed in 20 (10.7%) of the isolates. An analysis of the antifungal consumption data revealed that recent prior exposure to fluconazole increased the risk to be infected by a resistant strain. In particular, the duration of the treatment was significantly longer for patients infected by a resistant isolate, while the total and mean daily doses received did not impact the acquisition of resistance in C. glabrata No link between genotype and resistance was found. However, multilocus variable-number tandem-repeat analyses indicated a potential intrahospital spread of some isolates between patients. These isolates shared the same genetic profiles, and infected patients were hospitalized in the same unit during an overlapping period. Finally, quantitative real-time PCR analyses showed that, unlike that for other ABC efflux pumps, the expression of CgCDR1 was significantly greater in resistant strains, suggesting that it would be more involved in fluconazole (FLC) resistance. Our study provides additional evidence that the proper administration of fluconazole is required to limit resistance and that strict hand hygiene is necessary to avoid the possible spreading of C. glabrata isolates between patients.

Increasing candidaemia incidence from 2004 to 2015 with a shift in epidemiology in patients preexposed to antifungals.

Candidaemia is an important health problem in immunocompromised patients with an epidemiology varying with region, period and patient population involved. The occurrence of candidaemia and the associated species distribution over a 12-year period at a large tertiary care centre in Belgium were analysed. The trend in incidence in the intensive care units (ICUs) and non-ICUs was investigated as well as the influence of antifungal exposure on the species distribution. From 2004 until 2015, 865 candidaemia episodes occurred in 826 patients at the University Hospitals Leuven. Candida albicans (59%) remained the most important cause of candidaemia, followed by C. glabrata (22.4%) and C. parapsilosis (8%). The mean incidence in the whole hospital was 1.48 per 10 000 patient days (PD). The incidence in ICUs increased reaching up to 10.7 per 10 000 PD whereas in the non-ICUs, the incidence decreased. Prior exposure to fluconazole and echinocandins was associated with candidaemia caused by less susceptible species. Candidaemia incidence increased in the whole hospital, driven by ICUs. Surveillance of candidaemia epidemiology on a local scale is of high value to guide empirical treatment strategies.