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- Sampling protocol to assess Aspergillus section Fumigati in woodworking environmentsPublication . Dias, Marta; Gomes, Bianca; Pena, Pedro; Cervantes, Renata; Rodriguez, Margarida; Riesenberger, Bruna; Marques, Liliana; Ribeiro, Edna; Viegas, CarlaPortugal’s wood industry employs over 80,000 people and comprises about 16,600 enterprises in 2021. Sawmill workers are exposed to several occupational hazards, including microorganisms. Health problems have been documented by workers exposed to wood dust infected with fungi. A fungal priority pathogens list (WHO FPPL) was recently released by the WHO. It prioritizes fungal pathogens based on their perceived public health importance and unmet research and development needs. Some of the species on this list are frequently found in sawmills. Azole fungicides are commonly used by sawmills to protect wood. Aspergillus section Fumigati azole antifungal resistance has previously been observed in this environment. This study aimed to establish a sampling protocol to effectively isolate Aspergillus section Fumigati in woodworking environments.
- Budget-friendly protocol for TR34/L98H and TR46/Y121F/T289A mutation detection in Aspergillus section Fumigati isolatesPublication . Dias, Marta; Rodriguez, Margarida; Vasques, C.; Riesenberger, Bruna; Marques, Liliana; Gomes, Bianca; Pena, Pedro; Cervantes, Renata; Viegas, Susana; Viegas, CarlaAspergillus section Fumigati is one of the most common sections, in the environment. It has been found in different occupational environments, such as sawmills and waste sorting. Its cryptic species show intrinsic resistance to several antifungals. Resistance in A. fumigatus is emerging due to selective pressure caused by the prolonged use of azoles. It is often associated with mutations in the Cyp51A gene. The fungal priority pathogens list (WHO), includes A. fumigatus with critical priority. Further analysis to identify potential resistance mechanisms and mutations is needed. Objective: This evaluation aims to offer a protocol for mutation detection in Aspergillus section Fumigati isolates. It will contribute to the development of guidance that can support future occupational exposure assessments.
- 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.
- 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 contamination in Lisbon’s primary schools: sampling insights and analytical approachesPublication . Cervantes, Renata; Pena, Pedro; Gomes, Bianca; Dias, Marta; Riesenberger, Bruna; Rodriguez, Margarida; Marques, Liliana; Viegas, CarlaClimate change is posing challenges for Portugal due to intense weather changes, affecting public health and causing pathogens to adapt and spread, increasing the global risk of infectious diseases. Azole fungicides are less effective against resistant fungi, raising concerns for children. Warm and humid conditions promote the growth of pathogenic fungi and the production of mycotoxins, impacting health by causing gastrointestinal problems, organ damage, and chronic diseases Even after fungi removal, mycotoxins continue to pose risks. Objectives: identifying fungal species present in indoor environments; assessing the spatial distribution and concentration levels within classrooms and other areas; investigating factors influencing fungal proliferation, such as building characteristics and seasonal variations; evaluating the effectiveness of existing cleaning protocols and providing insights into proactive management strategies to protect students and staff members' health and well being.
- Aspergillus distribution on waste collection trucksPublication . Dias, Marta; Pena, Pedro; Cervantes, Renata; Gomes, Bianca; Caetano, Liliana Aranha; Viegas, CarlaIntroduction: While better waste management is viewed as a critical contributor to reducing health outcomes and harmful environmental impacts, microbiological occupational exposure in the waste management industry is often overlooked, resulting in detrimental health effects on employees. Some waste workers, such as garbage collectors and truck drivers transporting the residential waste, spend part of their shift in a truck cabin and part of it in a workplace where organic materials are processed. The aim of this study was to investigate the Aspergillus spp. contamination present in 32 waste collection trucks (WCT).
- 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.