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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.
- 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 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.
- Assessment of waste workers occupational risk to microbial agents and cytotoxic effects of mixed contaminants present in the air of waste truck cabin and ventilation filtersPublication . Marchand, Genevieve; Wingert, Loïc; Viegas, Carla; Aranha Caetano, Liliana; Viegas, Susana; Twaruzek, Magdalena; Lacombe, Nancy; Lanoie, Delphine; Valois, Isabelle; Gouin, Francois; Soszczyńska, Ewelina; Kosicki, Robert; Dias, Marta; Debia, MaximilienWorkers in the waste-processing industry are potentially exposed to high concentrations of biological contaminants, leading to respiratory and digestive problems and skin irritations. However, few data on the exposure of waste collection truck (WCT) drivers are available. The goal was to document the microbial risk of the waste collection truck (WCT) workers while in the vehicle cab. Long-period sampling using the truck air filters (CAF) and short-time ambient air sampling in the cab were used. The potential release of microbial particles from CAFs was also investigated since it could contribute to the microbial load of the cabin air. A combination of analytical methods also helped assess the complex mixture of the biological agents. Aspergillus sections Fumigati and Flavi, E. coli, Enterobacter spp., and Legionella spp. were detected in the CAF of trucks collecting three types of waste. The highest levels of bacteria and fungi were found in the CAF from organic WCT. The highest endotoxin concentrations in CAF were 300 EU/cm2. Most of the CAF showed cytotoxic effects on both lung cells and hepatocytes. Only one mycotoxin was detected in a CAF. The maximal concentrations in the ambient WCT air varied according to the type of waste collected. The highest proportion (84%) of the air samples without cytotoxic effects on the lung cells was for the recyclable material WCTs. The results revealed the potential microbial risk to workers from a complex mixture of bio-contaminants in the cabs of vehicles collecting all types of waste. The sustained cytotoxic effect indicates the potential adverse health-related impact of mixed contaminants (biological and non-biological) for the workers. Overall, this study highlights the benefits of using a complementary sampling strategy and combined analytical methods for the assessment of the microbial risk in work environments and the need to implement protective measures for the workers. Implications: Exposure to microbial agents is a well-known occupational hazard in the waste management sector. No previous study had evaluated the cytotoxicity of ambient air and ventilation filters to document worker exposure to a combination of contaminants during waste collection. This research confirms the usefulness of ventilation filters for the long-term characterization of exposure to infectious agents, azole-resistant fungi, coliform bacteria, and mycotoxin. Overall, this study highlights the importance of using several sampling and analysis methods for a comprehensive assessment of microbial risk in work environments, as well as the need to implement appropriate protective measures for collection workers.
- Unveiling the occupational exposure to microbial contamination in conservation-restoration settingsPublication . Viegas, Carla; Cervantes, Renata; Dias, Marta; Gomes, Bianca; Pena, Pedro; Carolino, Elisabete; Twarużek, Magdalena; Kosicki, Robert; Soszczyńska, Ewelina; Viegas, Susana; Caetano, Liliana Aranha; Pinheiro, Ana CatarinaAssuring a proper environment for the fulfillment of professional activities is one of the Sustainable Development Goals and is contemplated in the One Health approach assumed by the World Health Organization. This particular study is applied to an often neglected sector of our society—the conservators/restorers—despite the many health issues reported by these professionals. Three different specialties (textiles, paintings, and wood sculptures) and locations were selected for evaluation by the placement of electrostatic dust cloths. After treatment of the samples, bacterial and fungal contamination was assessed, as well as mycotoxin determination, the presence of azole-resistant strains, and cytotoxicity of the microorganisms encountered. Bacteria were only present in one of the media used and showed relatively low numbers. The highest level of contamination by fungi was identified in one of the textiles settings. The textile area also showed the highest variability for fungi. Aspergillus sp. is one indicator of possible environmental issues, and A. sections Fumigati and Circumdati were particularly relevant in two of the settings and identified in all of them. No mycotoxins were detected and the large majority of the fungi identified were non-cytotoxic. Overall, these can be considered low-contaminated environments but attention should be given to the Aspergillus sp. contamination. Additional studies are needed not only to make these results more robust but also to test if the environmental sampling alone is the best approach in a setting where there is very little movement and dust displacement and where professionals are in very close proximity to the artifacts being treated, which may suggest the existence of a micro-atmosphere worth evaluating and comparing to the obtained results.
- Exposure to azole-resistant Aspergillus fumigatus in Portuguese sawmillsPublication . Dias, Marta; Gomes, Bianca; Pena, Pedro; Cervantes, Renata; Caetano, Liliana Aranha; Carolino, Elisabete; Viegas, CarlaSawmill workers have an increased risk of adverse respiratory health effects due to wood dust, microorganisms, and metabolite exposure. Knowledge about exposure-response relationships is scarce, with information lacking regarding this transformation sector in Portugal. Among microbial components, fungi and fungal spores and fragments stand as allergens and irritants, due to their high prevalence, under the right circumstances, on wood products (planks, chips) that have been preserved. Aspergillus species can infect the lungs, causing pneumonia-like symptoms that can progress into more severe sickness and are reported as the most common found in sawmills. Moreover, Aspergillus section Fumigati has recently entered the WHO ranking list of most critical fungal pathogens, based on the impact of the fungal disease on public health. While antifungal resistance has been reported in sawmills, there is no data, to date, on occupational exposure to azole-resistant Aspergillus fumigatus (ARAF) in Portuguese sawmills. This study aims to determine the prevalence of azole-resistant Aspergillus fumigatus, followed by the characterization of the distribution of azole-resistance mutations in Aspergillus fumigatus from Portuguese sawmills. The correlation of the exposures with work tasks handled wood products, and seasonal variation will also be made to suggest the most suitable procedures to ensure sawmill workers’ safety.
- Microbiological contamination assessment in higher education institutesPublication . Viegas, Carla; Pimenta, Raquel; Dias, Marta; Gomes, Bianca; Brito, Miguel; Caetano, Liliana Aranha; Carolino, Elisabete; Gomes, Anita Q.The higher education sector represents a unique environment and it acts as a work environment, a learning environment for students, and frequently, also a home environment. The aim of this study was to determine the microbial contamination (SARS-CoV-2, fungi, and bacteria) in Higher Education Facilities (HEI) by using active and passive sampling methods and combining culture-based methods with molecular tools targeting Aspergillus section Fumigati. In addition, the resistance to azole profile was also assessed. Surface samples showed a range of total bacterial contamination between 1 × 103 to 3.1 × 106 CFU·m−2, while Gram-negative bacteria ranged from 0 to 1.9 × 104 CFU·m−2. Fungal contamination ranged from 2 × 103 to 1.8 × 105 CFU·m−2 on MEA and from 5 × 103 to 1.7 × 105 CFU·m−2 on DG18. The most prevalent species found on both media was Cladosporium sp. (47.36% MEA; 32.33% DG18). Aspergillus genera were observed on MEA (3.21%) and DG18 (14.66%), but not in the supplemented media used for the azole screening. Aspergillus section Fumigati was detected in 2 air samples (2.22%, 2 out of 90 samples) by qPCR. When testing for SARS-CoV-2 all results were negative. The present study showed that although cleaning and disinfection procedures are done regularly due to the COVID-19 pandemic, being effective in eliminating SARS-CoV-2, surfaces were often contaminated with microorganisms other than SARS-CoV-2. This can be a result of increasing resistance to biocides, and to the wide range of environmental factors that can contribute to the dissemination of microbial contamination indoors.
- Microbial assessment in a rare Norwegian book collection: a One Health approach to cultural heritagePublication . Sequeira, Sílvia O.; Pasnak, Ekaterina; Viegas, Carla; Gomes, Bianca; Dias, Marta; Cervantes, Renata; Pena, Pedro; Twarużek, Magdalena; Kosicki, Robert; Viegas, Susana; Caetano, Liliana Aranha; Penetra, Maria João; Silva, Inês; Caldeira, Ana Teresa; Pinheiro, CatarinaMicrobial contamination poses a threat to both the preservation of library and archival collections and the health of staff and users. This study investigated the microbial communities and potential health risks associated with the UNESCO-classified Norwegian Sea Trade Archive (NST Archive) collection exhibiting visible microbial colonization and staff health concerns. Dust samples from book surfaces and the storage environment were analyzed using culturing methods, qPCR, Next Generation Sequencing, and mycotoxin, cytotoxicity, and azole resistance assays. Penicillium sp., Aspergillus sp., and Cladosporium sp. were the most common fungi identified, with some potentially toxic species like Stachybotrys sp., Toxicladosporium sp., and Aspergillus section Fumigati. Fungal resistance to azoles was not detected. Only one mycotoxin, sterigmatocystin, was found in a heavily contaminated book. Dust extracts from books exhibited moderate to high cytotoxicity on human lung cells, suggesting a potential respiratory risk. The collection had higher contamination levels compared to the storage environment, likely due to improved storage conditions. Even though overall low contamination levels were obtained, these might be underestimated due to the presence of salt (from cod preservation) that could have interfered with the analyses. This study underlines the importance of monitoring microbial communities and implementing proper storage measures to safeguard cultural heritage and staff well-being.
- Cytotoxicity of Aspergillus section Fumigati isolated from health care environmentsPublication . Viegas, Carla; Twarużek, Magdalena; Almeida, Beatriz; Dias, Marta; Ribeiro, Edna; Carolino, Elisabete; Soszczyńska, Ewelina; Caetano, Liliana AranhaThis study analyzed 57 Aspergillus section Fumigati (AF) isolates collected by active and passive sampling (N = 450) in several health care facilities and from a biological sampling of health care workers (N = 25) and controls (N = 22) in Portugal. All isolates were cultured in different media and screened for azole resistance. Cytotoxicity was assessed for 40 isolates in lung epithelial cells and kidney cells using the MTT assay. Aspergillus section Fumigati was prevalent in the health care facilities and in nasal swabs from health care workers and controls. All AF isolates reduced cell viability and presented medium to high cytotoxicity, with cytotoxicity being significantly higher in A549 lung epithelial cells. The cytotoxicity of isolates from the air and nasal swab samples suggested the inhalation route as a risk factor. Notably, 42% of AF isolates exhibited a pattern of reduced susceptibility to some of the most used antifungals available for the treatment of patients infected with these fungi. In sum, the epidemiology and clinical relevance of Aspergillus section Fumigati should continue to be addressed. A deeper understanding of the mechanisms underlying Aspergillus-mediated cytotoxicity is necessary.
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