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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.
- 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.
- Preliminary results concerning school staff personal exposure to microbial load: worry to be considered?Publication . Pena, Pedro; Cervantes, Renata; Dias, Marta; Gomes, Bianca; Riesenberger, Bruna; Marques, Liliana; Rodriguez, Margarida; Viegas, CarlaWhy is it important to assess indoor air quality in schools? Is Portugal addressing educators’ health risks? Do the current guidelines cover school workers too? This study aims to provide a first insight into the personal exposure of teachers and school staff to bacteria and fungi in primary schools in Portugal.
- First insights of Portuguese primary schools’ fungal assessment: is indoor air quality legal framework suitable for this indoor setting?Publication . Pena, Pedro; Cervantes, Renata; Gomes, Bianca; Dias, Marta; Riesenberger, Bruna; Marques, Liliana; Sousa, Margarida; Viegas, CarlaThe assessment of microbial indoor air quality in schools is vital for promoting student health. Portugal's regulations focus on commercial buildings (Ordinance nº 138-G/2021), neglecting standards for schools. Evidence suggests indoor/outdoor fungal ratio inadequacies in high-risk areas like schools. Objective: To assess fungal threshold adequacy set by the Portuguese ordinance in different sites of schools located in the Lisbon area.
- A multi-approach sampling strategy to assess exposure to microbiologic agents in poultriesPublication . Gomes, Bianca; Dias, Marta; Pena, Pedro; Cervantes, Renata; Rodriguez, Margarida; Marques, Liliana; Riesenberger, Bruna; Viegas, CarlaA reasonable number of studies focusing on microbiological contamination associated with the poultry industry evidence various health concerns. In occupational studies focusing on microbiological contamination in poultry farms, air sampling is typically the only sampling method used. To better understand the relationship between factors influencing microbial contamination and adverse health effects, data regarding the amount, composition, and risk category of the common microorganisms are needed. This study intends to apply a multi-approach sampling protocol and corroborate the importance of its application for a wider microbial characterization.
- Preliminary results concerning school staff personal exposure to microbial load: worry to be considered?Publication . Pena, Pedro; Cervantes, Renata; Dias, Marta; Gomes, Bianca; Riesenberger, Bruna; Marques, Liliana; Rodriguez, Margarida; Viegas, CarlaSchool environments are potential exposure scenarios to bioaerosols. Breading such contaminants can contribute to the development of respiratory infection. Most studies performed in schools regarding air quality focus on children’s health, neglecting the personal exposure of teachers and school staff. This study aims to provide a first insight into the personal exposure of teachers and school staff to bacteria and fungi in primary schools in Portugal. Personal air samples were collected from teachers and staff with an SKC Button Aerosol Sampler (2 h sampling at 4 L/min flow rate) with a polycarbonate filter from 10 schools located in the Metropolitan Lisbon Area. Extracts were inoculated onto TSA and VRBA (7 days incubation at 30 °C and 37 °C, respectively) for bacterial quantification and onto MEA and DG18 (5–7 days incubation at 27 °C) for fungal quantification. Results showed higher bacterial contamination in teacher samples. Fungal counts followed the same pattern. Although bacterial and fungal counts were below recommended thresholds, the possible presence of potentially harmful pathogens requires qualitative assessment. These preliminary findings underscore the need for comprehensive strategies to address microbial contamination in schools, ensuring the safety and well-being of the school community.
- Levels of fungi in the air of poultry farms following different stages of birds’ growth cyclePublication . Gomes, Bianca; Dias, Marta; Pena, Pedro; Cervantes, Renata; Rodriguez, Margarida; Marques, Liliana; Riesenberger, Bruna; Carolino, Elisabete; Viegas, CarlaPoultry production intensification might influence microbial development. Concerning fungal assessment, despite the limited number of studies, several fungi have already been found in poultry pavilions. Occupational health specialists frequently overlook worker safety and biological risk assessment due to a lack of systematic knowledge regarding the agents involved. Consequently, workers, exposed to organic dust are at risk of developing various respiratory diseases. This study aimed to analyze indoor and outdoor fungal load at different stages of the poultries growth cycle. The sampling campaign was conducted in Winter in 14 poultry pavilions. Air samples from indoor (n = 47) and outdoor (n = 20) were taken before bird placement (sanitary control), followed by 1st, 2nd and 3rd weeks. The impactor MAS-100 was set at 100 L/min for 5 min. Fungal loads indoors exceed the outdoor in all stages of the production cycle. Besides, fungal occurrence was also identified during sanitary control (5%). The acquired data can provide new insights into the variation in fungal air concentrations during different phases of poultry growth. The data obtained should be complemented with qualitative research. Together, this information will support the development of mitigation techniques aiming to protect workers, animals, and environmental health and thus following a One Health approach.
- 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, CarlaAzole fungicides are commonly used in sawmills to protect wood against the wood-destroying basidiomycete fungi. Azole-resistant Aspergillus section Fumigati has previously been identified in these environments. Here we aimed to establish a sampling protocol to efficiently isolate Aspergillus section Fumigati in woodworking environments. The sampling campaign was conducted between December 2022 and November 2023 in 13 DIY stores and 6 sawmills. Both active and passive sampling methods were used. A total of 370 Aspergillus section Fumigati isolates were recovered from the accessed environments with particularly high efficiency using DG18 incubated at 37 °C. Both active and passive sampling media had a similar number of isolates. Regarding Andersen's six-stage air sampler, stage 5 had the highest number of isolates in both woodworking environments. Data presented in this study validated the used protocol to assess Aspergillus section Fumigati in woodworking environments to enable Occupational Health Services to prioritize interventions in these specific occupational environments to perform accurate exposure assessment and detailed risk assessment and management.
- Boosting knowledge on occupational exposure to microbial contamination in Portuguese carpentriesPublication . Dias, Marta; Gomes, Bianca; Pena, Pedro; Cervantes, Renata; Rodriguez, Margarida; Riesenberger, Bruna; Marques, Liliana; Carolino, Elisabete; Twarużek, Magdalena; Kosicki, Robert; Soszczynska. Ewelina; Caetano, Liliana Aranha; Viegas, Susana; Viegas, CarlaIntroduction: Wood industry workers face health risks due to exposure to microorganisms and their metabolites. This study aimed to characterize seasonal microbial contamination, antifungal resistance, mycotoxins, cytotoxicity, and particulate matter in Portuguese carpentries, to reduce exposure and promote safe working conditions. Methods: Conducted in six carpentries in Lisbon, Portugal, the sampling strategy encompassed active and passive sampling methods to assess microbial contamination. A Handheld Particle Counter HH3016-IAQ was used to monitor particulate matter size, temperature, and humidity. Results: The highest fungal load was in the cold season, with Aspergillus sp. being the predominant species, and the highest bacterial load was in the warm season. Reduced susceptibility to azoles was observed in both seasons, with greater species diversity in the cold season. In the warm season, Nidulantes and Fumigati sections of Aspergillus were detected by RT-PCR, with Fumigati being the most prevalent; in the cold season, only Nidulantes were detected. Mycotoxins, mainly fumonisins, were more prevalent in the warm season; in the cold season, griseofulvin was the most prevalent mycotoxin. Cytotoxicity was more prevalent in A549 cells than in SK cells. Settled dust caused greater cytotoxicity in SK cells, and filters from the vacuumed dust in A549 cells. Higher particulate matter concentrations in the sampled indoor areas suggest a significant contribution of indoor activities to workers’ exposure. Discussion: The study highlights concerns about seasonal variations in microbial contamination, emphasizing the potential for respiratory diseases, invasive infections by azole-resistant fungi, mycotoxin exposure, and cytotoxicity in lung cells due to co-exposure to fungi, particulate matter, and mycotoxins influenced by environmental conditions.