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- Portuguese cork industry: filling the knowledge gap regarding occupational exposure to fungi and related health effectsPublication . Viegas, Carla; Dias, Marta; Pacífico, Cátia; Faria, Tiago; Clérigo, Anália Matos; Dias, Hermínia Brites; Caetano, Liliana Aranha; Carolino, Elisabete; Gomes, Anita Q.; Viegas, SusanaIntroduction: The presence of the Penicillium section Aspergilloides (formerly known as Penicillium glabrum) in the cork industry involves the risk of respiratory diseases such as suberosis. Methods: This study aimed to corroborate the predominant fungi presents in this occupational environment by performing a mycological analysis of 360 workers’ nasal exudates collected by nasal swabs. Additionally, evaluation of respiratory disorders among the cork workers was also performed by spirometry. Results: Penicillium section Aspergilloides was detected by qPCR in 37 out of the 360 nasal swabs collected from workers’ samples. From those, 25 remained negative for Penicillium sp. when using culture-based methods. A significant association was found between ventilatory defects and years of work in the cork industry, with those people working for 10 or more years in this industry having an approximately two-fold increased risk of having ventilatory defects compared to those working less time in this setting. Among the workers who detected the presence of Penicillium section Aspergilloides, those with symptoms presented slightly higher average values of CFU. Discussion: Overall, the results obtained in this study show that working in the cork industry may have adverse effects on worker’s respiratory health. Nevertheless, more studies are needed (e.g., using serological assays) to clarify the impact of each risk factor (fungi and dust) on disease etiology.
- Tea contamination by mycotoxins and azole-resistant mycobiota: the need of a One Health approach to tackle exposuresPublication . Viegas, Carla; Simões, Ana Beatriz; Faria, Mariana; Gomes, Bianca; Cervantes, Renata; Dias, Marta; Carolino, Elisabete; Twaruzek, Magdalena; Kosicki, Robert; Viegas, Susana; Caetano, Liliana AranhaDespite tea beneficial health effects, there is a substantial risk of tea contamination by harmful pathogens and mycotoxins. A total of 40 tea samples (17 green (raw) tea; 13 black (fermented) tea; 10 herbal infusions or white tea) were purchased from different markets located in the Lisbon district in 2020. All products were directly available to consumers either in bulk (13) and or in individual packages (27). Bacterial analysis was performed by inoculating 150 μL of samples extracts in tryptic soy agar (TSA) supplemented with 0.2 % nystatin medium for mesophilic bacteria, and in Violet Red bile agar (VRBA) medium for coliforms (Gram-negative bacteria). Fungal research was performed by spreading 150 μL of samples in malt extract agar (MEA) supplemented with 0.05 % chloramphenicol and in dichloran-glycerol agar (DG18) media. The molecular detection of the Aspergillus sections Fumigati, Nidulantes, Circumdati, and Flavi was carried out by Real-Time PCR (qPCR). Detection of mycotoxins was performed using a high-performance liquid chromatograph (HPLC) with a mass spectrometry detector. Azole resistance screening was achieved following the EUCAST guidelines. The highest counts of total bacteria (TSA) were obtained in green raw tea (81.6 %), while coliform counts (VRBA) were found in samples from black raw tea (96.2 %). The highest fungal counts were obtained in green raw tea (87.7 % MEA; 69.6 % DG18). Aspergillus sp. was the most prevalent genus in all samples on MEA (54.3 %) and on DG18 (56.2 %). In the raw tea 23 of the samples (57.5 %) presented contamination by one to five mycotoxins in the same sample. One Aspergillus section Fumigati isolate from a green tea beverage recovered from itraconazole-Sabouraud dextrose agar (SDA) medium, presented itraconazole and posaconazole E-test MICs above MIC90 values. Our findings open further discussion regarding the One-Health approach and the necessary investment in researching biological hazards and azole resistance associated with the production and consumption of tea (in particular green tea).
- Commercial tea samples: a natural health ally or an Aspergillus infusionPublication . Gomes, Bianca; Dias, Marta; Cervantes, Renata; Caetano, Liliana Aranha; Viegas, CarlaIntroduction and Purpose - Tea consumption has been increasing in the last few years. The lack of specific legislation concerning microbial contamination of teas and infusions might represent a risk for consumers’ health. This study characterizes fungal contamination and azole resistance on green and black tea and other infusions commercially available in Portugal.
- Aspergillus spp. and azole-resistance characterization on mechanic protection gloves from waste sorting industryPublication . Viegas, Carla; Dias, Marta; Almeida, Beatriz; Caetano, Liliana AranhaAspergillus spp. species are filamentous fungi commonly found in different environmental compartments, where they thrive as saprophytes. Some species among the Aspergillus genera can be occasionally harmful to humans. Only a few species are considered as significant opportunistic pathogens in humans. Aspergillus section Fumigati is an opportunistic pathogen of immunocompromised hosts and one of several Aspergillus species that cause a wide range of respiratory disorders. Increased occurrence of opportunistic Aspergillus infections in immunocompromised patients, and the emergence of antifungal resistance, either in the clinical or in the environment had been reported.
- Produtos de saúde à base de plantas (módulo I)Publication . Caetano, Liliana AranhaProdutos de saúde à base de plantas: alimentos - chás medicinais; medicamentos à base de plantas; suplementos alimentares. Motivações para uso de produtos de saúde à base de plantas. Produtos de saúde à base de plantas vs medicamentos. Características dos produtos de saúde à base de plantas. O que os consumidores precisam saber? Medicamentos tradicionais à base de plantas. Enquadramento legal.
- Organic dust exposure in veterinary clinics: a case study of a small-animal practice in PortugalPublication . Viegas, Carla; Monteiro, Ana; Ribeiro, Edna; Caetano, Liliana Aranha; Carolino, Elisabete; Assunção, Ricardo; Viegas, SusanaLiterature about occupational health in small-animal veterinary practices is scarce, but most of it has recognised a number of risks to be considered, including organic dust exposure. The aim of this pilot study was to assess organic dust, bacterial, and fungal contamination in the indoor environment of a typical Portuguese veterinary clinic but also to screen for azoleresistant fungi. To complement these findings we also analysed workers’ nasal exudates for resistant bacteriota. Particles measurements included mass concentrations (PMC) of five particle sizes (PM0.5, PM1, PM2.5, PM5, PM10) and their counts (PNC). Indoor air samples were obtained from six locations as well as before and during cat dental cleaning and cultured on four media for bacterial and fungal assessment. An outdoor sample was also collected for reference Surface samples were taken from the same indoor locations using swabs and we also use electrostatic dust cloths as passive methods. PM10 showed the highest concentrations across the locations. Indoor air fungal loads ranged from 88 to 504 CFU m−3. The azole-resistant Aspergillus section Nigri was identified in one sample. Indoor air bacterial loads ranged from 84 to 328 CFU m-3. Nasopharyngeal findings in the 14 veterinary clinic workers showed a remarkably low prevalence of Staphylococcus aureus (7.1 %). Our results point to contamination with organic dusts above the WHO limits and to the need for better ventilation. Future studies should combine the same sampling protocol (active and passive methods) with molecular tools to obtain more accurate risk characterisation. In terms of prevention, animals should be caged in rooms separate from where procedures take place, and worker protection should be observed at all times.
- Poly(D,L-lactic acid) scaffolds as an innovative approach to the treatment of mixed S. aureus-C. albicans biofilmsPublication . Zegre, Miguel; Barros, J.; Ribeiro, I. A.; Santos, C.; Aranha Caetano, Liliana; Gonçalves, L.; Monteiro, F.; Ferraz, M.; Bettencourt, A.Introduction: The treatment of bacterial joint and bone infections in patients after multiple revision arthroplasties is very challenging. An expanding number of studies report the co-isolation of fungal and bacterial species (such as Candida albicans and Staphylococcus aureus) from polymicrobial biofilm associated with infections related to bone infections. Current investigations establish that local-specific drug delivery scaffolds with low toxicity and increased efficiency to specific sites when compared to oral and systemic administration approaches, can considerably lower the number of viable microorganisms in polymicrobial biofilms, preventing simultaneously the progression of infection in bone disorder. Notably, the development of co-delivery systems of at least two antimicrobials is yet a neglected approach, while it may be a critical strategy for the treatment of infections associated with polymicrobial biofilms. Simultaneously, it is recommended to assess the contribution of each microbial population within the biofilm to select the best therapy to treat polymicrobial infections. Among different biomaterials used in scaffolds as drug-delivery carriers, poly(lactic acid) (PLA) based polymers are being widely studied due to their versatility, low toxicity, and tailored biodegradability having the US Food and Drug Administration approval for clinical use. The adequate osteoconductive and anti-S. aureus effects of a collagen functionalized poly(D, L-lactic acid) (PDLLA) porous scaffold loaded with minocycline (a tetracycline antibiotic) have been previously demonstrated3. In the present study, we focus on the problem of mixed bacterial-fungal biofilm infections and the joining of two antimicrobials in the PDLLA scaffold. Minocycline and voriconazole (an antifungal triazole) were the chosen model drugs, since minocycline may represent a promising drug that can be administered in combination with azoles (namely voriconazole) to treat infections caused by pathogenic Candida species. Morphological and chemical properties of the co-delivery PDLLA scaffolds, as well as drug release profiles, were examined. The antibiofilm activity of these drug delivery systems was tested against single- and dual-species biofilms of S. aureus and C. albicans. The formation of dual-species S. aureus – C. albicans biofilms was studied over time to understand the relationship between both microorganisms during in vitro biofilm formation. Cytocompatibility and osteoconductive tests were also conducted using MG-63 osteoblasts to assess the biocompatibility of the PDLLA scaffolds.
- Assessment of azole resistance in clinical settings by passive samplingPublication . Aranha Caetano, Liliana; Almeida, Beatriz; Viegas, CarlaThe exposure to the fungal burden is an increased risk factor for the development of respiratory symptoms and fungal diseases. The emergence worldwide of azole resistance in fungal species is a major concern for public health. Clinical settings must provide a clean and safe environment to protect patients and staff from fungal diseases. The aim of this study was to assess the fungal burden and prevalence of azole resistance in clinical settings in Portugal using passive sampling methods. Ten Primary Health Care Centers were evaluated through passive sampling, including electrostatic dust cloths, heating ventilation, and air conditioning filters, and settled dust. All samples were plated onto Sabouraud dextrose agar media and screened for azole resistance using agar media supplemented with itraconazole, voriconazole and posaconazole through incubation at 27 °C, for 5 days. Fungal species were identified based on macro and micromorphology. Fungal load ranged from 348 to 424628 CFU·m−2 in EDC, 0 to 56500 CFU·m−2 in HVAC filter, and 2 to 514 CFU·g−1 in settled dust. EDC samples presented the highest fungal loads and wider diversity. Azole resistance was observed in all Primary Health Care Centers. Multi-azole resistance (fungal growth in two or more azoles) was observed in 90% units for Penicillium sp., C. sitophila, and Cladosporium sp. None of the samples containing Aspergillus sp. colonies presented resistance to the tested azoles. The knowledge of the fungal burden and prevalence of resistance to azole-based antifungal drugs in clinical environments will allow a better risk characterization regarding fungal burden.
- Aspergillus spp. prevalence in Primary Health Care Centres: assessment by a novel multi-approach sampling protocolPublication . Viegas, Carla; Almeida, Beatriz; Gomes, Anita Quintal; Carolino, Elisabete; Aranha Caetano, LilianaExposure to Aspergillus conidia may cause adverse effects on human health; however, no specific recommendations for routine assessments of Aspergillus in the clinical environment have been suggested so far. This study intended to determine the prevalence of Aspergillus in the clinical environment, focusing on ten Primary Health Care Centres (PHCC) through a novel multi-approach sampling protocol. Air and passive sampling, culture-based methods and a probe-based real-time assay for the detection of four clinically relevant Aspergillus sections were performed. Aspergillus spp. was observed in all PHCC, with highest prevalence on floor surface swabs (n=81) (18% on MEA; 6.94% on DG18). Regarding air samples (n=81), highest Aspergillus counts were found in the waiting room (94% MEA; 18% DG18), where Nigri was the most prevalent Aspergillus section. The use of a multi-approach sampling protocol to assess Aspergillus burden in the analyzed PHCC has greatly contributed to risk characterization, highlighting the need to implement corrective measures in order to avoid fungal presence in those settings.
- Occupational exposure to fungi and mycotoxins: overview of different studies developed in PortugalPublication . Viegas, Carla; Cervantes, Renata; Dias, Marta; Gomes, Bianca; Pena, Pedro; Carolino, Elisabete; Twarużek, Magdalena; Kosicki, Robert; Soszczyńska, Ewelina; Caetano, Liliana Aranha; Viegas, SusanaAs in all European countries, Portuguese employers are obliged by legal requirements to assess and prevent exposure to chemical, physical and biological risks (Directive 89/391/EEC). However, the health risks of biological origin are frequently less recognized and, thus, underreported. This study intends to present an overview of the assessments of fungal contamination and mycotoxins performed in three different occupational environments (Firefighters headquarters - FFH; Waste collection trucks – WCT and; Cemeteries - CM). Active sampling (impaction and impingement methods) and several passive sampling methods (swabs, settled dust, settled dust filters, electrostatic dust cloths – EDC, cleaning materials,…) were employed. Azole resistance screening, the molecular detection of Aspergillus sections, as well as mycotoxin analysis were also conducted. Regarding FFH it was raised concern regarding the following findings: a) microbial contamination didn´t comply with Portuguese IAQ legal requirements in most of the FFH (fungal load 45.45%); b) fungal exposure through inhalation underlining a possible risk factor for respiratory diseases; c) prevalence of Aspergillus sp. in cleaning materials that might represent a potential source of cross-contamination; d) widespread of Aspergillus section Fumigati in all the FFH analysed; e) trend of multidrug resistance found among fungi with a focus on Fumigati isolates; and f) detection of several mycotoxins (fumonisin B2, nivalenol, mycophenolic acid, and sterigmatocystin). Concerning WCT Aspergillus sp. (4.18%) was one of the most prevalent species. Aspergillus section Fumigati was detected in 5 samples. Mycotoxins were detected in filters (N=1) and in settled dust samples (N=16). The mycotoxin detected in the filter was fumonisin B1 and the most detected in settled dust was mycophenolic acid. Sterigmatocystin, deoxynivalenol, and zearalenon were detected in only one sample each. In what concerns CM, Aspergillus sp. presented the highest counts in DG18 (18.38%) and it wasn't observed in azole-supplemented SDA media. Mycophenolic acid was detected in one settled dust sample. Overall, our study reveals that a comprehensive sampling approach using active and passive sampling methods and combined analytic methods (i.e., culture-based and molecular) is an important asset in microbial exposure assessments. Furthermore, in all three occupational environments mycotoxins were detected unveiling this occupational health threat.