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- 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.
- Organic dust exposure in veterinary clinics: a case study of a small-animal practice in PortugalPublication . Viegas, Carla; Monteiro, Ana; Ribeiro, Edna; Caetano, Liliana Aranha; Carolino, Elisabete; Assunção, Ricardo; Viegas, SusanaLiterature about occupational health in small-animal veterinary practices is scarce, but most of it has recognised a number of risks to be considered, including organic dust exposure. The aim of this pilot study was to assess organic dust, bacterial, and fungal contamination in the indoor environment of a typical Portuguese veterinary clinic but also to screen for azoleresistant fungi. To complement these findings we also analysed workers’ nasal exudates for resistant bacteriota. Particles measurements included mass concentrations (PMC) of five particle sizes (PM0.5, PM1, PM2.5, PM5, PM10) and their counts (PNC). Indoor air samples were obtained from six locations as well as before and during cat dental cleaning and cultured on four media for bacterial and fungal assessment. An outdoor sample was also collected for reference Surface samples were taken from the same indoor locations using swabs and we also use electrostatic dust cloths as passive methods. PM10 showed the highest concentrations across the locations. Indoor air fungal loads ranged from 88 to 504 CFU m−3. The azole-resistant Aspergillus section Nigri was identified in one sample. Indoor air bacterial loads ranged from 84 to 328 CFU m-3. Nasopharyngeal findings in the 14 veterinary clinic workers showed a remarkably low prevalence of Staphylococcus aureus (7.1 %). Our results point to contamination with organic dusts above the WHO limits and to the need for better ventilation. Future studies should combine the same sampling protocol (active and passive methods) with molecular tools to obtain more accurate risk characterisation. In terms of prevention, animals should be caged in rooms separate from where procedures take place, and worker protection should be observed at all times.
- 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.
- Surveillance of mucorales resistance to azoles in the waste sorting industry: assessment of filtering respiratory protective devices and glovesPublication . Aranha Caetano, Liliana; Dias, M.; Almeida, Beatriz; Viegas, CarlaThe use of filtering respiratory protective devices (FRPD) and gloves is mandatory in the waste-sorting industry in Portugal as preventive of workers’ exposure to bioaerosols, including fungi. Mucorales order includes a large number of ubiquitous saprophytes species that can cause severe infections, such as mucormycosis, which is associated with a great deal of morbidity. The emergence of resistance to azoles among fungal species, including Mucorales, in the environmental challenges the management of severe fungal infections. The aim of this study was to assess the Mucorales burden retained by filtering respiratory protective devices (FRPD) (interior layer and exhalation valves) and present in gloves used in the waste sorting industry and to screen the resistance of Mucorales to azole-drugs in this occupational environment.
- 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).
- 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.
- Screening of fungal azole resistance in different environmental samplesPublication . Pena, Pedro; Morais, Joana; Caetano, Liliana Aranha; Viegas, CarlaFungi are responsible for skin diseases, infections of the lung and other organs, and bloodstream infections due to their presence in several environments such as in agriculture. Antimicrobial resistance has been described as a product of healthcare and agricultural overuse of azoles to inhibit fungal growth. The aim of this review article is to assess methodologies for fungi sampling and for the screening of azole resistance in environmental samples. The search was carried out in three search engines: PubMed, Scopus, and Web of Science (WoS). The Search terms in all databases were “fungal resistance in environmental samples”. After applying the inclusion criteria, the number of analyzed articles was 18 out of 71. The azole resistance tests were performed, in non-hospital environment samples, using mainly the EUCAST and CLSI M38-A2 broth microdilution methods. The most reported azole-resistant fungi were Aspergillus fumigatus, and the most common azoles used to perform the susceptibility tests were itraconazole, voriconazole, and posaconazole. Samples of air, water, and soil were considered, and the most found and studied fungi as Aspergillus fumigatus. This fungus is known to be an allergenic and mycotoxin producer with severe health consequences. It represents a concern for public health for its increasingly described resistance to the three azoles most used in clinical practice: itraconazole, voriconazole, and posaconazole. It is suggested the assessment of fungi in other occupational and indoor environments, as well as of their resistance for a better characterization of these environments. It is also suggested the use of complementary sampling methods for the fungal assessment.
- 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.
- 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.