Browsing by Author "Sabino, Raquel"
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- Accessing occupational exposure to fungi in a cork industryPublication . Viegas, C.; Clérigo, A.; Faria, T.; Sabino, Raquel; Veríssimo, Cristina; Quintal-Gomes, A.; Viegas, S.In this study we aimed to access fungal exposure in workers from one cork industry through the mycological analysis of their nasal exudate and the environmental fungal contamination of their surroundings as well. Nasal mucous samples from 127 workers were taken with sterilized cotton swabs.The fungal species identified in the collected nose swabs were shown to be correlated with the results obtained in the environment. Eighty workers (63.0%) presented contamination of their nose nostril with Chrysonilia sitophila, which number of colonies was countless. Within the Aspergillus genus, the complexes Fumigati, Circumdati, Versicolores and Candidi were isolated. No azole-resistant Aspergillus isolates grew in the selective media used (screened itraconazole and voriconazole resistance).This approach allowed us to estimate the risk associated with these tasks performance. Moreover, the cork industry is related to high dust contamination and this can promote exposure to fungi since dust particles can act as carriers of fungi to the worker’s nose. Assessment by molecular tools will ensure the specific targeting of DNA from P. glabrum complex in workers nose.
- Activity of Diphenyl diselenide against Aspergillus isolatesPublication . Melo, Aryse Martins; Poester, Vanice Rodrigues; Munhoz, Lívia; Trápaga, Mariana; Roca, Beatriz; Klafke, Gabriel Baracy; Sabino, Raquel; Stevens, David A.; Xavier, Melissa OrzechowskiOrganoselenium compounds have been showing promising antimicrobial activity against bacteria and some fungal species. Among these compounds, diphenyl diselenide (PhSe)2 is a simple and chemically stable molecule with proven low toxicity to animal hosts. Although the mechanisms of action of this molecule are not totally clear, it has been reported that it has a prooxidative activity for microorganisms, due to glutathione depletion. Given the emergence of azole resistance Aspergillus sp. isolates is a global and rising concern, research towards new molecules with antifungal potential are necessary. Thus, the aim of this study was to evaluate the in vitro susceptibility of Aspergillus spp. clinical isolates to (PhSe)2.
- Air fungal contamination in ten hospitals’ food units from LisbonPublication . Viegas, C.; Ramos, C.; Almeida, M.; Sabino, Raquel; Verissimo, Cristina; Rosado, L.A descriptive study was developed to monitor air fungal contamination in ten food units from hospitals. Fifty air samples of 250 litres were collected through impaction method. Samples were collected in food storage facilities, kitchen, food plating, canteen and also, outside premises, since this is the place regarded as reference. Simultaneously, environmental parameters were also monitored, including temperature and relative humidity through the equipment Babouc, LSI Sistems and according to the International Standard ISO 7726.
- Algorithm to assess the presence of Aspergillus fumigatus resistant strains: The case of Norwegian sawmillsPublication . Viegas, Carla; Almeida, Beatriz; Aranha Caetano, Liliana; Afanou, Anani; Straumfors, Anne; Veríssimo, Cristina; Gonçalves, Paulo; Sabino, RaquelAssociation between selection pressure caused by the use of azole fungicides in sawmills and the development of fungal resistance has been described. The aim of this study was to implement an algorithm to assess the presence of Aspergillus section Fumigati resistant strains in sawmills. Eighty-six full-shift inhalable dust samples were collected from eleven industrial sawmills in Norway. Different culture media were used and molecular identification to species level in Aspergillus section Fumigati was done by calmodulin sequencing and TR34/L98H and TR46/Y121F/T289A mutations were screened by real-time PCR assay and confirmed by cyp51A sequencing. Six Fumigati isolates were identified as A. fumigatus sensu stricto and two of these grew on azole-supplemented media and were further analyzed by real-time PCR. One was confirmed to be a TR34/L98H mutant. The obtained results reinforce the need to assess the presence of A. fumigatus sensu stricto resistant isolates at other workplaces with fungicide pressure.
- Algorithm to assess the presence of Aspergillus fumigatus resistant strains: The case of Norwegian sawmillsPublication . Viegas, Carla; Almeida, Beatriz; Aranha Caetano, Liliana; Afanou, Anani; Straumfors, Anne; Veríssimo, Cristina; Gonçalves, Paulo; Sabino, RaquelAssociation between selection pressure caused by the use of azole fungicides in sawmills and the development of fungal resistance has been described. The aim of this study was to implement an algorithm to assess the presence of Aspergillus section Fumigati resistant strains in sawmills. Eighty-six full-shift inhalable dust samples were collected from eleven industrial sawmills in Norway. Different culture media were used and molecular identification to species level in Aspergillus section Fumigati was done by calmodulin sequencing and TR34/L98H and TR46/Y121F/T289A mutations were screened by real-time PCR assay and confirmed by cyp51A sequencing. Six Fumigati isolates were identified as A. fumigatus sensu stricto and two of these grew on azole-supplemented media and were further analyzed by real-time PCR. One was confirmed to be a TR34/L98H mutant. The obtained results reinforce the need to assess the presence of A. fumigatus sensu stricto resistant isolates at other workplaces with fungicide pressure.
- Algoritmo para o diagnóstico laboratorial das infeções fúngicas profundasPublication . Sabino, Raquel; Simões, Helena; Veríssimo, CristinaAmostras de tecidos de doentes com suspeita de infeções fúngicas profundas ou subcutâneas foram analisadas no Laboratório Nacional de Referência de Infeções Parasitárias e Fúngicas do INSA, de acordo com um algoritmo de diagnóstico desenvolvido que apresenta uma abordagem polifásica, que tem por objetivo permitir um diagnóstico laboratorial mais rápido das infeções fúngicas profundas. Quarenta e seis amostras de tecido de 39 doentes com suspeita de infeções fúngicas profundas ou subcutâneas foram analisadas durante um período de 26 meses ( janeiro de 2015-fevereiro de 2017) pelo laboratório do INSA, usando uma abordagem laboratorial que incluiu cultura, PCR panfúngica e PCR dirigida a Aspergillus. Do total de amostras estudadas, 23 foram positivas para fungos (PCR, cultura e/ou histologia). Das 46 amostras, 16 apresentaram resultados positivos para DNA fúngico. Em 12 amostras foi detetado um sinal positivo por PCR panfúngica e em 6 por PCR dirigida a Aspergillus (em 2 das amostras ocorreu deteção pelas duas metodologias). Em 61% (22/36) das amostras estudadas, houve concordância entre os métodos moleculares e culturais. Os agentes etiológicos identificados foram Candida albicans, C. glabrata, C. tropicalis, Trichosporon montevideense, Alternaria spp., Exophiala sp., Trichoderma sp., Histoplasma spp., Aspergillus fumigatus, Trichophyton rubrum e Paracoccidioides brasiliensis. Os resultados obtidos mostraram que a abordagem polifásica proposta parece ser uma boa estratégia na deteção de fungos em amostras de tecidos, permitindo eventualmente um melhor prognóstico. Em estudos posteriores, pretende-se estudar um maior número de amostras clinicas e implementar mais metodologias moleculares que permitam a deteção dirigida a determinados géneros fúngicos.
- Analysis of Aspergillus spp. burden by culture and molecular based methods in different occupational environments: what needs to be done?Publication . Quintal Gomes, Anita; Faria, Tiago; Caetano, Liliana Aranha; Sabino, Raquel; Viegas, CarlaFungal burden have traditionally being detected by conventional culture analysis, which despite its limitations, is widely used by the scientific community. Alternatively, quantitative real-time PCR (qPCR), based on the amplification of genomic regions specific to certain fungal species, has been associated with increased sensivity, allowing the detection of dormant forms of fungi, such as spores. We present several studies where both methods were used to detect the presence of toxigenic fungi, namely Aspergillus, particularly from the Fumigati, Flavi and Circumdati sections
- Analysis of clinical and environmental Candida parapsilosis isolates by microsatellite genotyping—a tool for hospital infection surveillancePublication . Sabino, Raquel; Sampaio, P.; Rosado, L.; Videira, Z.; Grenouillet, F.; Pais, C.Candida parapsilosis emerged as an important opportunistic pathogen, causing candidaemia worldwide. Nosocomial outbreaks triggered by this species have been frequently described, particularly in cancer patients. For a better understanding of its epidemiology, several typing methods are used and microsatellite analysis has been reported as highly discriminant. The main objective of this work was to study C. parapsilosis isolates by application of microsatellite genotyping to distinguish epidemiologically related strains, compare clinical and environmental isolates and determine possible routes of dispersion of the isolates in the hospital setting. A total of 129 C. parapsilosis isolates from different origins, including hospital environment and hands of healthcare workers, were genotyped using four microsatellite markers. The isolates were recovered from different health institutions. Analysis of C. parapsilosis isolates from hospital environment showed great genotypic diversity; however, the same or very similar genotypes were also found. The same multilocus genotype was shared by isolates recovered from the hand of a healthcare worker, from the hospital environment and from patients of the same healthcare institution, suggesting that these could be possible routes of transmission and that infections due to C. parapsilosis may be mainly related with exogenous transmission to the patient. Examination of sequential isolates from the same patients showed that colonizing and bloodstream isolates had the same multilocus genotype in the majority of cases. We demonstrate that this typing method is able to distinguish clonal clusters from genetically unrelated genotypes and can be a valuable tool to support epidemiologic investigations in the hospital setting.
- 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.
- Anti-Aspergillus fumigatus IgG in patients with bronchiectasis and its relationship with clinical outcomePublication . Rodrigues Trápaga, Mariana; Rodrigues Poester, Vanice; Sanchotene, Karine Ortiz; Martins Melo, Aryse; Benelli, Jéssica Louise; Basso, Rossana Patricia; Klafke, Gabriel Baracy; Fernandes Ramos, Daniela; Veríssimo, Cristina; Sabino, Raquel; Stevens, David A.; Xavier, Melissa OrzechowskiAspergillosis is a mycosis, most commonly afecting the airways. This mycosis can worsen the clinical condition of patients with concurrent lung diseases. We assayed for the presence of serum anti-A. fumigatus IgG in bronchiectasis patients from a tertiary hospital in south Brazil and evaluated the relationship with clinical outcome. Thirty-one patients with bronchiec tasis, without cystic fbrosis, were included. Clinical and epidemiological data were collected from all participants. Positive serological tests were detected in 13% (4/31) of the patients. The mortality rate for the year following the assay was, in the seropositive group, 75% (3/4), whereas in the seronegative group, 15% (4/27). An illustrative case is also shown and discussed. Our study highlights the diagnostic challenge and the possible impact of Aspergillus infection on these patients, indicating the necessity of more and larger investigations in the feld