Percorrer por autor "Twarużek, Magdalena"
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- 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.
- Aspergillus spp. in woodworking settings: implications for occupational health and safetyPublication . Dias, Marta; Rodriguez, Margarida; Riesenberger, Bruna; Marques, Liliana; Carolino, Elisabete; Twarużek, Magdalena; Kosicki, Robert; Gonçalves, Lídia; Caetano, Liliana Aranha; Alastruey-Izquierdo, Ana; Viegas, Susana; Viegas, CarlaWoodworkers are exposed to several potentially harmful agents, including microorganisms that grow in the wood. The most common fungal species in woodworking environments are Aspergillus, Penicillium, and Cladosporium spp., with occupational exposure to Aspergillus spp. posing a significant respiratory risk. This study aimed to assess exposure to Aspergillus spp. in two Portuguese woodworking environments and to perform a thorough analysis of Aspergillus fumigatus complex isolates collected from 13 DIY stores and 6 Carpentries in the Lisbon Metropolitan Area. Sampling combined active and passive methods to assess microbial contamination. Aspergillus fumigatus isolates were analysed for their antifungal susceptibility, resistant mechanisms, mycotoxin production, and cytotoxic potential in lung (A459) and liver (HepG2) cell lines. The MAS-100 presented Aspergillus sections Aspergilli and Flavi with the highest prevalence in DIY stores and Carpentries, respectively. A total of 1185 Aspergillus spp. were recovered, 270 identified as Aspergillus fumigatus sensu stricto growing at 37 °C. None of those isolates was resistant to azoles, 99.07% of them produced gliotoxin, and 39.9% of them produced cytotoxic effects in at least one cell line. This study comprehended a multi-approach that considered not only sampling methods but also the laboratory assays to be applied in the Aspergillus section Fumigati isolates recovered from two different woodworking environments, allowing a complete and robust analysis of this specific environment and species. Overall, the findings indicate that woodworkers are exposed to A. fumigatus isolates with relevant pathogenic traits, despite the absence of azole resistance, underscoring the need for continued environmental and occupational monitoring.
- Assessment of Portuguese fitness centers: bridging the knowledge gap on harmful microbial contamination with focus on fungiPublication . Viegas, Carla; Peixoto, Cátia; Gomes, Bianca; Dias, Marta; Cervantes, Renata; Pena, Pedro; Slezakova, Klara; Pereira, Maria do Carmo; Morais, Simone; Carolino, Elisabete; Twarużek, Magdalena; Viegas, Susana; Caetano, Liliana AranhaThe lack of knowledge regarding the extent of microbial contamination in Portuguese fitness centers (FC) puts attendees and athletes at risk for bioaerosol exposure. This study intends to characterize microbial contamination in Portuguese FC by passive sampling methods: electrostatic dust collectors (EDC) (N=39), settled dust (N=8), vacuum filters (N=8), and used cleaning mops (N=12). The obtained extracts were plated in selective culture media for fungi and bacteria. Filters, EDC, and mop sample extracts were also screened for antifungal resistance and used for the molecular detection of the selected Aspergillus sections. The detection of mycotoxins was conducted using a high-performance liquid chromatograph (HPLC) system and to determine the cytotoxicity of microbial contaminants recovered by passive sampling, HepG2 (human liver carcinoma) and A549 (human alveolar epithelial) cells were employed. The results reinforce the use of passive sampling methods to identify the most critical areas and identify environmental factors that influence microbial contamination, namely having a swimming pool. The cardio fitness area presented the highest median value of total bacteria (TSA: 9.69x102 CFU.m-2.day-1) and Gram-negative bacteria (VRBA: 1.23 CFU.m-2.day-1), while for fungi it was the open space area, with 1.86x101 CFU.m-2.day-1. Aspergillus sp. was present in EDC and in filters used to collect settled dust. Reduced azole susceptibility was observed in filters, EDC (on ICZ and VCZ), and mops (on ICZ). Fumonisin B2 was the only mycotoxin detected and was present in all sampling matrixes except settled dust. High and moderate cytotoxicity was obtained, suggesting that A549 cells were more sensitive to samples’ contaminants. The observed widespread of critical toxigenic fungal species with clinical relevance, such as Aspergillus section Fumigati, as well as Fumonisin B2 emphasizes the importance of frequent and effective cleaning procedures while using shared mops appeared as a vehicle of cross-contamination.
- Assessment of the microbial contamination in 'Do It Yourself' (DIY) stores: a holistic approach to protect workers’ and consumers’ healthPublication . Dias, Marta; Gomes, Bianca; Pena, Pedro; Cervantes, Renata; Gonçalves, Sara; Carolino, Elisabete; Twarużek, Magdalena; Kosicki, Robert; Ałtyn, Iwona; Caetano, Liliana Aranha; Viegas, Susana; Viegas, CarlaIntroduction: In “Do-It-Yourself” (DIY) stores, workers from the wood department are considered woodworkers. Given the health risks associated with woodworking, particularly from fungi and their metabolites, this study aims to assess microbial contamination and health risks for both workers and customers. Methods: The study was developed in 13 DIY stores in the Lisbon Metropolitan Area, Portugal. It employed a comprehensive sampling approach combining active (MAS-100, Andersen six-stage, Coriolis μ, and SKC Button Aerosol Sampler) and passive (electrostatic dust collectors, surface swabs, e-cloths, settled dust, filters from vacuumed dust, filtering respiratory protection devices, and mechanical protection gloves) methods to assess microbial contamination. A Lighthouse Handheld Particle Counter HH3016- IAQ was used to monitor the particulate matter size, temperature, and humidity. Results: The wood exhibition area presented the highest fungal load, while the payment area exhibited the highest bacterial load. MAS-100 detected the highest fungal load, and surface swabs had the highest bacterial load. Penicillium sp. was the most frequently observed fungal species, followed by Aspergillus sp. Mycotoxins, namely mycophenolic acid, griseofulvin, and aflatoxin G1, were detected in settled dust samples and one filter from the vacuum cleaner from the wood exhibition area. Cytotoxicity evaluation indicates the wood-cutting area has the highest cytotoxic potential. Correlation analysis highlights relationships between fungal contamination and particle size and biodiversity differences among sampling methods. Discussion: The comprehensive approach applied, integrating numerous sampling methods and laboratory assays, facilitated a thorough holistic analysis of this specific environment, enabling Occupational and Public Health Services to prioritize interventions for accurate exposure assessment and detailed risk management.
- Assessment of the microbial contamination of mechanical protection gloves used on waste sorting industry: a contribution for the risk characterizationPublication . Viegas, Carla; Twarużek, Magdalena; Dias, Marta; Almeida, Beatriz; Carolino, Elisabete; Kosicki, Robert; Soszczyńska, Ewelina; Grajewski, Jan; Aranha Caetano, Liliana; Viegas, SusanaIn Portugal, mechanical protection gloves (MPG) are of mandatory use and during their use sweat is released and, consequently, the humidity of the material increases leading to conditions favorable to the growth of microorganisms. However, no studies have been conducted in MPG to assess the bioburden. This study intended to determine the bioburden present in MPG and their biological effects and to discuss the possibility to use MPG as a passive method to assess occupational exposure to microbial contamination. Fungal burden was characterized through molecular tools for fungal toxigenic species, and antifungal resistance and mycotoxin profiles were determined. Cell viability was determined in swine kidney (SK) monolayer and hepatocellular carcinoma (Hep G2) cell lines. All MPG samples presented Gram-negative bacteria. The fungal contamination ranged from 0 CFU.m−2 in both MEA and DG18, to 5.09 × 106 and 2.75 × 106, and the most common fungi found was Aspergillus spp. (50.46%). Azole resistant Aspergillus sections were found in azole supplemented media. Aspergillus sections (Circumdati, Flavi, Fumigati and Versicolores) were detected by molecular tools in 66 out of 67 samples. The most-reported mycotoxin was mycophenolic acid (89.6%). HepG2 cells appear to be more sensitive to MPG contamination, with high cytotoxicity (IC50 < 0.05 mm2/ml) observed for 18 out of 57 gloves. MPG can be used in passive sampling to assess occupational exposure to bioburden in waste sorting industries and contribute to risk characterization. Some contaminants of MPG had cytotoxic potential and affected the biology of hepatic cells more than renal cells.
- Beyond quantitative indicators: comprehensive characterization of indoor fungal contamination in Portuguese elementary schoolsPublication . Cervantes, Renata; Pena, Pedro; Carolino, Elisabete; Twarużek, Magdalena; Kosicki, Robert; Viegas, Susana; Viegas, CarlaChildren are particularly vulnerable to environmental pollutants, making indoor air quality in schools a key health determinant. This study addresses regulatory gaps and the lack of standardized methods for assessing fungal contamination in Portuguese elementary schools, particularly regarding pathogenicity, toxigenicity, and antifungal resistance. Fungal contamination was assessed in 11 Portuguese elementary schools using surveys on building context and operation, active and passive air sampling, molecular detection of Aspergillus sections by qPCR, mycotoxin and azole resistance screening, and PM measurements. Results: Schools showed heterogeneous ventilation, with some relying on natural ventilation and others using mechanical systems. Airborne fungal contamination was dominated by respirable fractions, reaching 3.3×102 CFU m-3. Culture-based analyses revealed distinct fungal profiles, with Cladosporium sp. and Penicillium sp. frequently prevailing, while Aspergillus detection increased at higher incubation temperatures. Seasonal variability was observed, with contamination patterns differing between seasons. Rural and urban schools showed contrasting fungal distributions, with higher concentrations in rural schools, particularly in EDCs on DG18 at 27 °C (1.67 × 103 CFU/m2). Azole resistance screening detected resistant Aspergillus sections across environments. In gymnasiums, the Fumigati section represented 100% of sections detected on ITZ in filters and EDCs. In classrooms, Circumdati and Fumigati accounted for 100% of sections detected on ITZ in EDCs, while the Candidi section represented 100% in filters. The Fumigati section also grew on VOZ and POZ in EDCs. Mycotoxins, including ochratoxin A, mycophenolic acid, and sterigmatocystin, were detected in 1.93% of dust samples. Particle concentrations occasionally exceeded reference thresholds in warm (47.1% PM2.5; 58.8% PM10) and cold seasons (47.4% PM2.5; 23.1% PM10). Regulatory thresholds did not prevent the detection of relevant fungal hazards, including Aspergillus species and azole-tolerant growth. These findings show that quantitative indicators alone may not capture exposure complexity, highlighting the need for complementary data and the limitations in the current legal framework.
- Bioburden assessment by passive methods on a clinical pathology service in one central hospital from Lisbon: what can it tell us regarding patients and staff exposure?Publication . Viegas, Carla; Twarużek, Magdalena; Lourenço, Raquel; Dias, Marta; Almeida, Beatriz; Aranha Caetano, Liliana; Carolino, Elisabete; Gomes, Anita Q.; Kosicki, Robert; Soszczyńska, Ewelina; Viegas, SusanaThe assessment and control of microbial contamination in healthcare facilities is presently a mandatory and vital part of strategies to prevent and control hospital-acquired infections. This study aims to assess the bioburden with two passive sampling methods (30 ventilations grids swabs and 16 electrostatic dust collectors (EDCs)) at Clinical Pathology Services. The fungal burden was characterized through molecular tools, antifungal resistance, and the mycotoxins and cytotoxicity profile. Total bacteria presented the highest prevalence in both matrixes, whereas Gram-bacteria presented the lowest. Swabs presented a higher prevalence (27.6%) for fungal burden. Chrysonilia sitophila presented the highest prevalence in swabs, whereas for EDCs, C. sitophila and Mucor sp. were the most prevalent. Concerning Aspergillus genera on swabs, section Flavi was the one with the highest prevalence (58.02%), whereas, for EDCs, section Versicolores was the only section observed (100%). Aspergillus section Fumigati was detected in 10 swabs and 7 EDC samples and Aspergillus section Versicolores were detected in one EDC sample. Fungal growth on azole-supplemented media was observed in eight EDC samples. No mycotoxins were detected in any of the samples. A low cytotoxic effect was observed in two sites upon incubation of collected samples with A549 and SK cells and in two other sites upon incubation of collected samples with SK cells only. A medium cytotoxic effect was observed with one EDC sample upon incubation with A549 cells. This study reinforces the need for determination of the azole resistance profile for fungal species and allowed a preliminary risk characterization regarding the cytotoxicity. An intervention including the use of ultraviolet with a wavelength between 200 nm and 280 nm (UVC)—emitting device and increased maintenance and cleaning of the central heating, ventilation, and air conditioning (HVAC) systems should be ensured to promote the reduction of microbial contamination.
- Bioburden contamination and Staphylococcus aureus colonization associated with firefighter's ambulancesPublication . Viegas, Carla; Sousa, Pedro; Dias, Marta; Caetano, Liliana Aranha; Ribeiro, Edna; Carolino, Elisabete; Twarużek, Magdalena; Kosicki, Robert; Viegas, SusanaAmbulance vehicles are an essential part of emergency clinical services. Bioburden control in ambulances, thorough cleaning and disinfection, is crucial to minimize hospital-acquired infections, cross contamination and exposure of patients and ambulances‘ crew. In Portugal, firefighter crews are responsible, besides fire extinction, for first aid and urgent pre-hospital treatment. This study assessed the bioburden in Portuguese firefighters’ ambulances with a multi-approach protocol using active and passive sampling methods. Fungal resistance profile and mycotoxins detection in ambulances' ambient, and S. aureus (SA) prevalence and resistance profile in ambulances' ambient and colonization in workers were also investigated. Toxigenic fungi with clinical relevance, namely Aspergillus section Fumigati, were found on ambulance's air in the hazardous dimension range. Interestingly, surface contamination was higher after cleaning in several sampling sites. Prevalence of S. aureus was 3% in environmental samples, of which 2% were methicillin-sensitive (MSSA) and 1% methicillin-resistant (MRSA). About 2.07 fungal species were able to grow in at least one azole, ranging from one (44% samples) to five (6% samples) species in each azole. Mycotoxins were detected in mops and electrostatic dust cloths. Colonization by S. aureus in the firefighter crew was observed with a high associated prevalence, namely 48%, with a 24% prevalence of MSSA (8/33) and 21% of MRSA (7/33). Additional studies are needed to determine the potential risk of infection transmission between different vehicle fleets and under varying conditions of use. This will strengthen the paramedic sector's mission to save lives without putting their own health and safety at risk.
- Bioburden in health care centers: is the compliance with Portuguese legislation enough to prevent and control infection?Publication . Viegas, Carla; Almeida, Beatriz; Monteiro, Ana; Aranha Caetano, Liliana; Carolino, Elisabete; Gomes, Anita Quintal; Twarużek, Magdalena; Kosicki, Robert; Marchand, Geneviéve; Viegas, SusanaThis study intends to assess the bioburden in ten Portuguese health care centers (PHCC) with a multi-approach protocol using active (air sampling by impaction and impinger) and passive (surface swabs and air-conditioning filters) sampling methods, and compare the results with compliance levels of IAQ Portuguese legislation for the assessment of bioburden. The fungal burden in the PHCC was also characterized through molecular detection of toxigenic species, antifungal resistance and mycotoxins profile. Concerning legal compliance criteria for bacteriota the first criteria [indoor] + 350 CFU. m−3 < [outdoor] was not compiled in all PHCC analyzed. Regarding fungal load, 60% did not comply with the quantitative guideline (I/O < 1). Growth of four fungal genera (Chrysosporium, Cladosporium, Mucor, and Penicillium) was observed in HVAC filter samples collected from 6 out of 10 assessed PHCC when using azole-supplemented SDA media. Overall, Aspergillus/Penicillium/Paecilomyces levels were above the detection limits, except in one PHCC. The presence of mycotoxins was found both in air and HVAC filter samples. From the air samples analyzed, nine were contaminated (ng/ml) with 1–5 different mycotoxins within the same sample, whereas in HVAC filters, four samples contained 1 or 2 mycotoxins in the same filter. Overall, it was possible to conclude that Portuguese legislation is not enough to ensure IAQ in health care settings. The multi-approach sampling protocol used in this study allowed to unveil a more real scenario regarding exposure to bioburden. Detection of mycotoxins reinforces the relevance of studying mycotoxins in the clinical environment.
- 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.