Browsing by Author "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.
- 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.
- 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.
- Characterization of occupational exposure to fungal burden in Portuguese bakeriesPublication . Viegas, Carla; Faria, Tiago; Aranha Caetano, Liliana; Carolino, Elisabete; Quintal-Gomes, Anita; Twarużek, Magdalena; Kosicki, Robert; Viegas, SusanaSeveral studies reported adverse respiratory health effects in workers exposed to ambient contaminants in bakeries. The aim of this study was to examine worker exposure to fungi and mycotoxins in Portuguese bakeries in order to develop new policies in occupational health. Environmental samples such as air, surfaces, settled dust and electrostatic dust collector (EDC) were collected in 13 bakeries for fungal and mycotoxins assessment. Air samples obtained by impaction were performed applying malt extract agar (MEA) supplemented with chloramphenicol (0.05%) and dichloran glycerol (DG18) agar-based media. Air samples collected through impinger method were determined as well for fungal detection by molecular tools of Aspergillus sections and mycotoxins. The highest median value for fungal load was 1053 CFU·m-3 and 65.3% (32 out of 49) of the sampling sites displayed higher fungal load than limits imposed by the World Health Organization. Aspergillus genera was found in air, surface swabs and EDC. Molecular tools were effective in measuring Aspergillus section Fumigati in 22.4% on air, 27.8% on surface swabs and in 7.4% in EDC and Aspergillus section Versicolores in one air sample. All settled dust samples showed contamination with six to eight mycotoxins in each sample. The mycotoxins detected were deoxynivalenol-3-glucoside, deoxynivalenol, zearalenone, 15-acetyldeoxynivalenol, monoacetoxyscirpenol, diacetoxyscirpenol, fumonisin B1, fumonisin B2, griseofulvin, HT2, ochratoxin A, ochratoxin B and mycophenolic acid. Industrial hygienists and exposure assessors should rely on different sampling methods (active and passive) and different assays (culture based and molecular methods) to obtain an accurate risk characterization regarding fungal burden (fungi and mycotoxins). Additionally, the awareness for the raw material as a potential mycotoxins indoor contamination source is important.
- Characterization of the bioburden in green tea samples marketed in LisbonPublication . Viegas, Susana; Twarużek, Magdalena; Kosicki, Robert; Grajewski, Jan; Santos, Patrícia; Sá, Flávio; Mateus, Margarida; Viegas, CarlaTea is one of the most consumed beverages in the world essentially because of its beneficial effects on health. There are no regulations or load limits concerning microbial contamination of teas, and for this reason, microbiological food safety hazards linked with these products have rarely been reported. The aim of this study was to assess the bioburden of loose and bags tea of green tea marketed in Lisbon, after and before boiling following the ISO 7218 (2007) and ISO 3103 (1980). Twenty samples of green tea from loose and bag tea (ten of each) were select from different markets in Lisbon. For microbiological evaluation was carried out the extraction of the samples before and after the boiling. For samples that did not have manufacturer confection indications were followed by ISO 7218_2007. The samples were inoculated on two media: dichloro-glycerol agar (DG18) and Rose Bengal Chloramphenicol (RBC) incubated for 5 days at 25ºC. All tea samples will be screened for mycotoxins presence. Regarding fungal contamination in tea bags, before boiling, ranged from 0 to 66.67 CFU.g-1 in DG18, and from 0 to 84.85 CFU.g-1 in RBC medium. In tea bags, all brew samples present 0 CFU.mL-1 for the two medium. For loose tea before boiling, the fungal contamination ranged from 0 to 96.97 CFU.g-1, and after boiling ranged from 0 to 30.3 CFU.mL-1 in DG18. In RBC, before boiling, the fungal concentration ranged from 0 to 72.73 CFU.g-1 and the brew samples presented 0 CFU.mL-1 or were below the limit of determination after boiling. In loose tea samples, before boiling, the DG18 media show five different fungal genera/species and the most prevalent were Chrysosporium sp. (38.46%), Aspergillus section Nigri (30.76%) and section Versicolores (11.5%), whereas in RBC media two different fungal species were found, Aspergillus section Nigri (80%) and Penicillium sp. (20%). After boiling only the DG18 presented fungal contamination: Mucor sp. (80%) and Chrysonilia sitophila (20%). Concerning tea bags samples, before boiling, Aspergillus section Nigri was the most prevalent fungi in both media (90.47% DG18; 90.62% RBC). None of the bag samples showed fungal contamination after boiling. This study showed that after boiling was observed reduction of viable microorganisms, revealing that boiling is effective to reduce most bioburden present in the tea before boiling. However, due to the presence of toxigenic species in the tea before boiling and the well-known mycotoxins thermal resistance, mycotoxins data will be of interest to consider a more accurate risk assessment.
- Comprehensive assessment of occupational exposure to microbial contamination in waste sorting facilities from NorwayPublication . Viegas, Carla; Eriksen, Elke; Gomes, Bianca; Dias, Marta; Cervantes, Renata; Pena, Pedro; Carolino, Elisabete; Twarużek, Magdalena; Aranha Caetano, Liliana; Viegas, Susana; Graff, Pål; Afanou, Anani Komlavi; Straumfors, AnneIntroduction: It is of utmost importance to contribute to filling the knowledge gap concerning the characterization of occupational exposure to microbial agents in the waste sorting setting (automated and manual sorting). Methods: This study intends to apply a comprehensive field sampling and laboratory protocol (culture-based methods and molecular tools), assess fungal azole resistance, as well as to elucidate potential exposure-related health effects (cytotoxicity analyses). Skin-biota samples (swabs) were performed on workers and controls to identify other exposure routes. Results: In personal filter samples the guidelines in one automated industry surpassed the guidelines for fungi. Seasonal influence on viable microbial contamination including fungi with reduced susceptibility to the tested azoles was observed, besides the observed reduced susceptibility of pathogens of critical priority (Mucorales and Fusarium sp.). Aspergillus sections with potential toxigenic effects and clinical relevance were also detected in all the sampling methods. Discussion: The results regarding skin biota in both controls´ and workers´ hands claim attention for the possible exposure due to hand-to-face/mouth contact. This study allowed the conclusion that working in automated and manual waste sorting plants implies high exposure to microbial agents.