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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).
- Assessing the impact of climate change on indoor fungal contamination in Lisbon Metropolitan Area primary schools: a comprehensive studyPublication . Cervantes, Renata; Pena, Pedro; Dias, Marta; Gomes, Bianca; Viegas, CarlaThe increasing occurrence of severe weather events caused by global climate change raises concerns about indoor fungi. These changes can potentially alter fungal communities, promoting resistant strains, and intensifying mycotoxin production, having significant implications for human health. In educational settings, inadequate ventilation and high moisture levels amplify indoor fungal growth and mycotoxin contamination, posing a health risk that contributes to respiratory illnesses and allergic reactions in children and school staff. Our research explores the complex relationship between climate change and fungal diseases, highlighting the importance of temperature and moisture in driving fungal growth and mycotoxin production. This project focuses on the Lisbon metropolitan area and aims to assess the indoor levels of contamination and human exposure to azole-resistant fungi and mycotoxins in primary schools. The objective of this investigation is to carry out a comprehensive microbial characterization of the exposure to fungal contamination in primary educational institutions and to achieve that goal a comprehensive sampling campaign was employed, by the use of active and passive sampling methods and material collection from schools. Air samples utilizing the MAS-100 device, which collected 400L at a flow rate of 200 L/min, and the Anderson six-stage device, which collected 200L at a flow rate of 28.3 L/min were performed. Additionally, we collected samples of mops and surface swabs, and samples of settled dust through vacuuming and EDC, located in the sampling sites at 1.5 m height for 30 days, and collected during approximately 30 days. The samples will be analysed by culture-based methods, through the inoculation onto two different culture media: malt extract agar (MEA) supplemented with chloramphenicol (0.05%) incubated at 27ºC and dichloran-glycerol agar (DG18), incubated at 27ºC and 37ºC for 6 days. We will also be performing molecular detection of the selected fungal sections (Aspergillus sections Circumdati, Flavi, Fumigati, and Nidulantes). To evaluate the degree of mycotoxin contamination in school environments, we will employ high-performance liquid chromatography (HPLC) to detect the presence of 38 different mycotoxins. The preliminary results that will be presented are specifically related to the active air sampling. The ultimate objective is to provide essential insights for the development of effective risk management strategies that safeguard the health of school children and personnel in the face of growing fungal threats.
- 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.
- Bioburden assessment in Lisbon groceriesPublication . Moreira, Sílvia; Dias, Marta; Gomes, Bianca; Cervantes, Renata; Pena, Pedro; Viegas, CarlaObjective: This study aims to characterize the occupational exposure to the bioburden of groceries workers and to identify the most critical workstations. Background To our knowledge, this is the first study performed in Portugal that intends to characterize microbial contamination in this specific setting. Method: This study was conducted between November and December of 2020 in fifteen groceries stores (M) located in the municipality of Cascais. Passive sampling methods (Electrostatic dust cloths—EDC) and surface swabs were used at three different locations (checkout, fruits/vegetables, and warehouse/dispenser areas). EDC and surface swabs were inoculated in four standard culture media, namely, malt extract agar (MEA), dichloran glycerol (DG18), tryptic soy agar (TSA), and violet red bile agar (VRBA) for further characterization. Results: The prevalence of bacterial contamination was higher than fungal contamination in both sampling methods. The highest prevalence was in the fruits/vegetable area regarding fungal and bacterial contamination. Conclusion: The sampling methods employed effectively identified the most critical workplaces regarding microbial contamination. Application This study will be useful for industrial hygienists since it is the first held in small grocery shops focusing on the occupational exposure assessment of microbiological contamination.
- One Health approach in “Do It Yourself” stores to tackle fungal contaminationPublication . Dias, Marta; Gomes, Bianca; Pena, Pedro; Cervantes, Renata; Viegas, Susana; Viegas, CarlaWood dust contamination by fungi poses health risks due to exposure by inhalation. This study aimed to assess the exposure of woodworking employees and customers to fungal load in 13 “Do It Yourself” (DIY) stores in the Lisbon metropolitan area (Portugal). An active sampling method (MAS-100 air sampler) was applied in different store areas, collecting 200L at a flow rate of 140 L/min. Regarding the occupational exposure assessment and considering the threshold suggested by the WHO of 150 CFU.m-3, the cutting area and the wood display surpass this limit in all the stores included. As for the threshold suggested by OSHA for fungi in non-industrial workplaces (1.0 x 101-1.0 x 104 CFU/m³), all the sampling sites from all the stores are within the limit, although indoor/outdoor ratio showed a higher indoor load compared to the outdoor counts in most of the stores. Regarding the Portuguese Indoor Air Quality legal framework, although the ratio I/O complied in two out of 13 stores, it was possible to identify one of these two toxigenic species with a quantitative cut-off above the legal frame. The results of this assessment raise health concerns regarding workers´ and customers ‘safety and support the need to implement measures to prevent exposure and protect workers' and customers' health. Additionally, it is known that wood dust and shavings from DIY stores may be used in animal bedding and agriculture, promoting fungal contamination in other settings. This highlights the need for a One Health approach.
- First insights of Portuguese primary schools’ fungal assessment: is indoor air quality complying with Portuguese legal framework?Publication . Pena, Pedro; Cervantes, Renata; Gomes, Bianca; Dias, Marta; Viegas, CarlaThe assessment of Microbial Indoor Air Quality in elementary schools is essential to create healthier school environments and improve children's health outcomes. Portugal only established protection thresholds for microbial exposure, in commercial and service buildings through Portaria nº 138-G/2021, leaving schools neglected for such parameters. Since this legislation has already been proven insufficient for fungal parameters (based on indoor/outdoor ratio) in other critical environments such as healthcare centers, this study aims to assess fungal load compliance in different sites of schools located in the Lisbon area. An active sampling method was applied by collecting air through a MAS-100 device collecting 200L at a flow rate of 100 L/min. Of the 10 schools assessed in a summer campaign, 9 did not comply with the Portuguese legal framework in at least one site (8 out of 10 in the classrooms, 5 out of 7 in the bathroom, 4 out of 9 in the canteen, 4 out of 6 in the gymnasiums, and 3 out of 8 in the library). A critical assessment was performed to identify fungal species listed in this legislation. Although one school complies with the quantitative cut-off (I/O), critical species such as Aspergillus section Circumdati (Aspergillus ochraceus), and Aspergillus section Fumigati (Aspergillus fumigatus) present in this school do not meet the toxigenic species quantitative cut-off. The risk of exposure to pathogenic fungi poses a major public health threat for school-age children impacting not only treatable fungal infections but also students' learning conditions and outcomes.
- Fungal and mycotoxins contamination in non-alcoholic beverages: the case of tea from Portuguese market and coffee beans from BrazilPublication . Viegas, Carla; Gomes, Bianca; Cervantes, Renata; Dias, Marta; Pena, Pedro; Oliveira, Filipe; Andrade, Ednilton Tavares de; Caetano, Liliana Aranha; Twaruzek, Magdalena; Kosicki, Robert; Viegas, SusanaAgriculture constitutes one of the most sensitive sectors that could be affected by climate change. Among the xenobiotics contaminating agricultural crops, fungi and mycotoxins are the most challenging since their presence represents an economic burden due to crop loss and serious health effects related to animals and humans with severe repercussions. This study intends to characterize the fungal and mycotoxin contamination in tea available in the Portuguese market and in coffee from two Brazilian industries.
- Insights of Portugal’s teaching staff microbial occupational exposurePublication . Pena, Pedro; Cervantes, Renata; Gomes, Bianca; Dias, Marta; Viegas, CarlaEU strategies in education focus on risk factor awareness to promote a safe and healthy environment for the community. Health and safety promotion faces challenges due to the lack of adjusted policies for teaching staff. Teachers may be exposed to biohazards in schools, posing a health risk, which remains unexplored. Effective preventive measures and risk management actions require a thorough risk assessment. This study aims to conduct a microbial assessment regarding school staff personal exposure to bacterial and fungal loads.
- Portuguese groceries: a potential indoor environment to be used as public health sentinel for fungal and mycotoxins contaminationPublication . Viegas, Carla; Cervantes, Renata; Gomes, Bianca; Moreira, Silvia; Dias, Marta; Pena, Pedro; Carolino, Elisabete; Twaruzek, Magdalena; Kosicki, Robert; Caetano, Liliana Aranha; Viegas, SusanaThis study aimed to characterize microbial contamination in Portuguese grocery stores. The research was carried out in 15 grocery stores in Cascais, Portugal. Electrostatic dust cloths (EDC) and surface swabs were used on 3 sampling sites: checkout, fruits/vegetables, and warehouse/dispenser. Fungal contamination was characterized by culture-based methods, through the inoculation onto two different culture media: malt extract agar (MEA) supplemented with chloramphenicol (0.05 %) and dichloran-glycerol agar (DG18). Screening of azole resistance was performed by inoculation of EDC extracts' in azole-supplemented Sabouraud dextrose agar (SDA) media, according to EUCAST guidelines. A total of 39 samples were screened for mycotoxin contamination. The highest fungal contamination was obtained from fruits/vegetables in swabs (76% MEA; 71% DG18). The most prevalent genera in swabs was Cladosporium sp. (54% MEA). Regarding EDC, besides Penicillium sp. (63%) the most prevalent were Aspergillus sections Circumdati (25%) and Aspergilli (35%). Penicillium sp. was highly prevalent in the checkout (61% SDA; 87% voriconazole), and warehouse/dispenser sectors (73% voriconazole; 52% posaconazole). Circumdati and Fumigati were the most prevalent Aspergillus sections in fruits/vegetables (73% and 64% in voriconazole, respectively). Four mycotoxins (fumonisin B1, B2, B3) were detected simultaneously in most of the 36 samples. Identifying the most critical workplaces in groceries concerning fungal and mycotoxin contamination is of utmost importance to human health (workers and consumers) when using a One Health approach.