Browsing by Author "Gomes, Anita Quintal"
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- Analysis of surfaces for characterization of fungal burden - Does it matter?Publication . Viegas, Carla; Faria, Tiago; Meneses, Márcia; Carolino, Elisabete; Viegas, Susana; Gomes, Anita Quintal; Sabino, RaquelMycological contamination of occupational environments can be a result of fungal spores' dispersion in the air and on surfaces. Therefore, it is very important to assess it in both types of the samples. In the present study we assessed fungal contamination in the air and in the surface samples to show relevance of surfaces sampling in complementing the results obtained in the air samples.
- Exposure assessment in one central hospital: A multi-approach protocol to achieve an accurate risk characterizationPublication . Viegas, Carla; Almeida, Beatriz; Monteiro, Ana; Paciência, Inês; Rufo, João; Aguiar, Lívia; Lage, Bruna; Diogo Gonçalves, Lídia Maria; Caetano, Liliana Aranha; Carolino, Elisabete; Gomes, Anita Quintal; Twarużek, Magdalena; Kosicki, Robert; Grajewski, Jan; Teixeira, João Paulo; Viegas, Susana; Pereira, CristianaThe bioburden in a Hospital building originates not only from patients, visitors and staff, but is also disseminated by several indoor hospital characteristics and outdoor environmental sources. This study intends to assess the exposure to bioburden in one central Hospital with a multi-approach protocol using active and passive sampling methods. The microbial contamination was also characterized through molecular tools for toxigenic species, antifungal resistance and mycotoxins and endotoxins profile. Two cytotoxicity assays (MTT and resazurin) were conducted with two cell lines (Calu-3 and THP-1), and in vitro pro-inflammatory potential was assessed in THP-1 cell line. Out of the 15 sampling locations 33.3% did not comply with Portuguese legislation regarding bacterial contamination, whereas concerning fungal contamination 60% presented I/O > 1. Toxigenic fungal species were observed in 27% of the sampled rooms (4 out of 15) and qPCR analysis successfully amplified DNA from the Aspergillus sections Flavi and Fumigati, although mycotoxins were not detected. Growth of distinct fungal species was observed on Sabouraud dextrose agar with triazole drugs, such as Aspergillus section Versicolores on 1 mg/L VORI. The highest concentrations of endotoxins were found in settled dust samples and ranged from 5.72 to 23.0 EU.mg-1. While a considerable cytotoxic effect (cell viability < 30%) was observed in one HVAC filter sample with Calu-3 cell line, it was not observed with THP-1 cell line. In air samples a medium cytotoxic effect (61-68% cell viability) was observed in 3 out of 15 samples. The cytokine responses produced a more potent average cell response (46.8 ± 12.3 ρg/mL IL-1β; 90.8 ± 58.5 ρg/mL TNF-α) on passive samples than air samples (25.5 ± 5.2 ρg/mL IL-1β and of 19.4 ± 5.2 ρg/mL TNF-α). A multi-approach regarding parameters to assess, sampling and analysis methods should be followed to characterize the biorburden in the Hospital indoor environment. This study supports the importance of considering exposure to complex mixtures in indoor environments.
- Fungal burden in waste industry: an occupational risk to be solvedPublication . Viegas, Carla; Faria, Tiago; dos Santos, Mateus; Carolino, Elisabete; Gomes, Anita Quintal; Sabino, Raquel; Viegas, SusanaHigh loads of fungi have been reported in different types of waste management plants. This study intends to assess fungal contamination in one waste-sorting plant before and after cleaning procedures in order to analyze their effectiveness. Air samples of 50 L were collected through an impaction method, while surface samples, taken at the same time, were collected by the swabbing method and subject to further macro- and microscopic observations. In addition, we collected air samples of 250 L using the impinger Coriolis μ air sampler (Bertin Technologies) at 300 L/min airflow rate in order to perform real-time quantitative PCR (qPCR) amplification of genes from specific fungal species, namely Aspergillus fumigatus and Aspergillus flavus complexes, as well as Stachybotrys chartarum species. Fungal quantification in the air ranged from 180 to 5,280 CFU m(-3) before cleaning and from 220 to 2,460 CFU m(-3) after cleaning procedures. Surfaces presented results that ranged from 29×10(4) to 109×10(4) CFU m(-2) before cleaning and from 11×10(4) to 89×10(4) CFU m(-2) after cleaning. Statistically significant differences regarding fungal load were not detected between before and after cleaning procedures. Toxigenic strains from A. flavus complex and S. chartarum were not detected by qPCR. Conversely, the A. fumigatus species was successfully detected by qPCR and interestingly it was amplified in two samples where no detection by conventional methods was observed. Overall, these results reveal the inefficacy of the cleaning procedures and that it is important to determine fungal burden in order to carry out risk assessment.
- Prevalence of Aspergillus fumigatus complex in waste sorting and incineration plants: an occupational threatPublication . Viegas, Carla; Gomes, Anita Quintal; Faria, Tiago; Sabino, RaquelAspergillus fumigatus is one of the major ubiquitous saprophytic fungi and it is considered one of the fungal species with higher clinical relevance. This study aimed at characterising the prevalence of A. fumigatus complex in one waste-sorting plant and also in one incineration plant. Conventional and molecular methodologies were applied in order to detect its presence. Aspergillus fumigatus complex was the second most frequently found in the air from the waste-sorting plant (16.0%) and from the incineration plant (18.0%). Regarding surfaces, it ranked the third species most frequently found in the waste-sorting plant (13.8%) and the second in the incineration plant (22.3%). In the waste-sorting plant, it was possible to amplify by qPCR DNA from the A. fumigatus complex in all culture-positive sampling sites plus one other sampling site that was negative by culture analysis. Considering the observed fungal load, it is recommended to apply preventive and protective measures in order to avoid or minimise worker's exposure.