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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.
- 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.
- 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.
- Are workers from waste sorting industry really protected by wearing filtering respiratory protective devices? The gap between the myth and realityPublication . Viegas, Carla; Dias, Marta; Almeida, Beatriz; Aranha Caetano, Liliana; Carolino, Elisabete; Gomes, Anita Quintal; Twarużek, Magdalena; Kosicki, Robert; Grajewski, Jan; Marchand, Geneviève; Viegas, SusanaOne of the solutions for decreasing the workers' exposure to bioburden is the use of Filtering Respiratory Protective Devices (FRPD). As such it is important to determine whether these devices are fulfilling their protective role. This is the basis of the current study, aimed at characterizing bioburden retained by 120 FRPD (both in interior layers and in exhalation valves) through culture based-methods and molecular tools and also via analysis of antifungal resistance and mycotoxins profile. Our results show that Gram–Bacteria are present at a higher prevalence than total bacteria in both matrixes. Regarding fungal identification, Chrysonilia sitophila presented the highest prevalence on interior layers (55.1% on malt extract agar (MEA) supplemented with chloramphenicol (0.05%); 59.6% on dichloran-glycerol agar (DG18)), whereas on exhalation valves Aspergillus sp. presented the highest prevalence on MEA (6.8%) and C. sitophila on DG18 (36.3%). Among Aspergillus genera, section Fumigati was the one with the highest prevalence in both matrices. Aspergillus sp. was the most prevalent on exhalation valves (75.0% ITRA) in the screening of azole resistance. Fumigati section was the most abundant Aspergillus sp. detected on the interior layers (33.33%, 40 samples out of 120) and on the exhalation valves (1.66%, 2 samples out of 120). The interior layers and exhalation valves from workers with more waste contact showed an increased exposure to bioburden. This study showed that FRPD can have high levels of bioburden, toxigenic fungal strains and Aspergillus sections with reduced susceptibility to the tested azoles and can be used as a passive sampling method since it mimics the results obtained by active methods in previous studies. The gathered information will be useful to prioritize multiple interventions on workers’ education or even on FRPD replacement frequency.
- 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.