Browsing by Author "Gomes, Anita Quintal"
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- Accessing occupational exposure to fungi in a cork industryPublication . Viegas, Carla; Clérigo, Anália; Faria, Tiago; Sabino, Raquel; Gomes, Anita Quintal; Viegas, SusanaDifferent forms of fungal diseases affecting the nose and paranasal sinuses are recognized, including invasive and non-invasive fungal rhinosinusitis. Penicillium glabrum complex is associated with respiratory diseases such as suberosis, a typical disease of cork industry workers. In addition, Chrysonilia sitophila has been described as causing occupational asthma, associated to prolonged exposure to high counts of spores. In this study we aimed to access fungal exposure in workers from one cork industry through the mycological analysis of their nasal exudate and the environmental fungal contamination of their surroundings as well.
- Analysis of Aspergillus spp. burden by culture based-methods and molecular methods in different occupational environments: what needs to be done?Publication . Gomes, Anita Quintal; Faria, Tiago; Caetano, Liliana Aranha; Sabino, Raquel; Viegas, CarlaAim of the study - Fungal burden has traditionally been detected by conventional culture analysis, which despite its limitations, is widely used by the scientific community. Alternatively, quantitative real-time PCR (qPCR), based on the amplification of genomic regions specific to certain fungal species, has been associated with increased sensivity, allowing the detection of dormant forms of fungi, such as spores. We present several studies where both methods were used to detect the presence of toxigenic fungi, namely Aspergillus, particularly from the Fumigatus Flavi and Circumdati sections.
- Analysis of fungal burden by conventional and molecular methods in different settings and matrices: implications for public and occupational healthPublication . Gomes, Anita Quintal; Faria, Tiago; Caetano, Liliana Aranha; Carolino, Elisabete; Viegas, Susana; Viegas, CarlaFungal burden has traditionally been detected by conventional culture analysis. This method allows the identification and quantification of organisms posing high health/occupational risk and is widely used by the scientific community. The fungal burden determined by culture analysis can, in most studies, be compared with legal and scientific guidelines allowing an estimation of the degree of severity of the exposure. However, this method is limited by several factors, including incubation conditions such as the incubation time, which can be very long for some species, thus preventing a quick assessment of fungal burden. Another limiting factor is competition between species: clinically relevant species might possess lower growth rates than other non-toxic fast growing fungi thus hampering their detection. These limitations can be overcome by the use of quantitative real-time PCR (qPCR). This method, based on the amplification of genomic regions specific to certain fungal species, increases sensitivity, allowing the specific detection of a given species and removing interference by other species present in the sample. qPCR also allows the detection of dormant forms of fungi, such as spores. Thus the ideal scenario is to use these the two methods in parallel, as they complement each other to provide useful information for the assessment of exposure to fungi. We briefly describe several studies where both methods were used to detect the presence of toxigenic fungi, namely Aspergillus, particularly from the Fumigati, Flavi and Circumdati sections. These include fungal analysis from different matrices such as air, feed and coffee and within different settings, including wastewater treatment plants, slaughterhouses, feed industries, poultry and swine pavilions. The results obtained with both conventional and molecular methods are compared and discussed as well as its implications for the exposed workers' health.
- Analysis of surfaces for characterization of fungal burden: does it matter?Publication . Viegas, Carla; Faria, Tiago; Meneses, Márcia; Carolino, Elisabete; Viegas, Susana; Gomes, Anita Quintal; Sabino, RaquelObjectives: Mycological contamination of occupational environments can be a result of fungal spores’ dispersion in the air and on surfaces. Therefore, it is very important to assess it in both types of the samples. In the present study we assessed fungal contamination in the air and in the surface samples to show relevance of surfaces sampling in complementing the results obtained in the air samples. Material and Methods: In total, 42 settings were assessed by the analysis of air and surfaces samples. The settings were divided into settings with a high fungal load (7 poultry farms and 7 pig farms, 3 cork industries, 3 waste management plants, 2 wastewater treatment plants and 1 horse stable) and a low fungal load (10 hospital canteens, 8 college canteens and 1 maternity hospital). In addition to culture-based methods, molecular tools were also applied to detect fungal burden in the settings with a higher fungal load. Results: From the 218 sampling sites, 140 (64.2%) presented different species in the examined surfaces when compared with the species identified in the air. A positive association in the high fungal load settings was found between the presence of different species in the air and surfaces. Wastewater treatment plants constituted the setting with the highest number of different species between the air and surface. Conclusions: We observed that surfaces sampling and application of molecular tools showed the same efficacy of species detection in high fungal load settings, corroborating the fact that surface sampling is crucial for a correct and complete analysis of occupational scenarios.
- 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 prevalence in air conditioning filters from vehicles: taxis for patient transportation, forklifts, and personal vehiclesPublication . Viegas, Carla; Moreira, Ricardo; Faria, Tiago; Caetano, Liliana Aranha; Carolino, Elisabete; Gomes, Anita Quintal; Viegas, SusanaThe frequency and importance of Aspergillus infections is increasing worldwide. This study aimed to assess the occupational exposure of forklifts and taxi drivers to Aspergillus spp. Nineteen filters from air conditioning system of taxis, 17 from forklifts and 37 from personal vehicles were assessed. Filters extract were streaked onto MEA, DG18 and in azole-supplemented media. Real-time quantitative PCR amplification of selected Aspergillus species-complex was also performed. Forklifts filter samples presented higher median values. Aspergillus section Nigri was the most observed in forklifts filters in MEA (28.2%) and in azole-supplemented media. DNA from Aspergillus sections Fumigati and Versicolores was successfully amplified by qPCR. This study enlightens the added value of using filters from the air conditioning system to assess Aspergillus spp. occupational exposure. Aspergillus azole resistance screening should be included in future occupational exposure assessments.
- Aspergillus spp. burden on filtering respiratory protective devices: is there an occupational health concern?Publication . Viegas, Carla; Dias, Marta; Almeida, Beatriz; Carolino, Elisabete; Gomes, Anita Quintal; Viegas, SusanaThere is scientific evidence that associates occupational exposure to fungi in the waste industry with health outcomes, being Aspergillus genera reported as the most prevalent. During Filtering Respiratory Protective Devices (FFR) use, humidity and temperature conditions provide favorable conditions for Aspergillus growth. The aim of this study was to characterize the Aspergillus burden retained by the FFR interior layer and exhalation valve. The sample consisted of 120 FFR and the correspondent interior layer and exhalation valves collected from different workstations from one waste sorting industry. The fungal content of exhalation valve and from the interior layer of the each FFR was extracted and inoculated on malt extract agar (MEA) supplemented with chloramphenicol (0.05%) and dichloran-glycerol agar (DG18). FFR extracts were also used for the molecular detection of Aspergillus sections Flavi and Fumigati. Valves presented Aspergillus spp. as the most prevalent genera on MEA media (44.4%), unlike what was observed in the FFR interior layer, where Aspergillus spp. was not the most prevalent genera (6.84% MEA; 5.2% DG18). Among the Aspergillus genera, section Fumigati was the one with the highest prevalence, both in exhalation valves (76.57% MEA; 87.24% DG18) and interior layer (75.81% MEA; 51.22% DG18). qPCR analysis successfully amplified DNA from the Aspergillus sections Flavi and Fumigati. Interior layers presented statistically significant differences showing a higher number of FFR contaminated with higher levels of Aspergillus in the workstations where workers contact directly with waste. This work, revealing strong fungal contamination of FFR used by workers at the waste industry, clearly indicates that future trials to test FFR protective efficacy and establishment of times for FFR replacement should be performed.
- Aspergillus spp. prevalence and azole-resistance surveillance on waste sorting industryPublication . Viegas, Carla; Faria, Tiago; Caetano, Liliana Aranha; Gomes, Anita QuintalAim of the study - The genus Aspergillus is one of the most prevalent in several highly contaminated occupational environments, such as the waste sorting industry. The identification and quantification of Aspergillus sections present on filters from the air-conditioning systems in fork lifters from waste industry was performed, including the screening of azole-resistance. In addition, toxigenic strains from Aspergillus spp. were detected.
- Aspergillus spp. prevalence in different Portuguese occupational environments: what is the real scenario in high load settings?Publication . Viegas, Carla; Faria, Tiago; Caetano, Liliana Aranha; Carolino, Elisabete; Gomes, Anita Quintal; Viegas, SusanaThe genus Aspergillus is one of the most prevalent regarding fungi in several highly contaminated occupational environments. The goal of the current study was to assess the prevalence of Aspergillus spp. in different settings, focusing on those where a higher load of fungal contamination is expected according to the European Agency for Safety and Health at Work. A specific protocol to ensure a more accurate assessment of the exposure to Aspergillus spp. is proposed aimed at allowing a detailed risk characterization and management. Two wastewater treatment plants, one wastewater elevation plant, four waste treatment plants, three cork industries, five slaughterhouses, four feed industries, one poultry pavilion, and two swineries, all located in the outskirts of Lisbon, were assessed. In total, 125 air samples and 125 surface samples were collected and analyzed by culture-based methods. Real-time polymerase chain reaction was performed to detect the fungal presence in 100 samples, targeting the Aspergillus sections Circumdati, Flavi, and Fumigati. The highest prevalence of Aspergillus spp. was found in wastewater treatment plants (69.3%; 31.1%), waste treatment plants (34.8%; 73.6%), and poultry feed industry (6.3%; 26.1%), in air and surfaces, respectively. Aspergillus spp. was also prevalent in cork industry (0.9%; 23.4%), slaughterhouses (1.6%; 17.7%), and swineries (7.4%; 9.5%), in air and surfaces, respectively. The Aspergillus sections more prevalent in the air and surfaces of all the assessed settings were the Nigri section (47.46%; 44.71%, respectively), followed by Fumigati (22.28%; 27.97%, respectively) and Flavi (10.78%; 11.45%, respectively) sections. Aspergillus section Fumigati was successfully amplified by qPCR in 18 sampling sites where the presence of this fungal species had not been identified by conventional methods. It should be highlighted that the occupational exposure burden is due not only to the Aspergillus load but also to the toxigenic potential of this genus. Based on our results, a protocol relied on the application of conventional and molecular methods in parallel is herein suggested aimed at allowing a better risk characterization and management.
- Aspergillus spp. prevalence in Primary Health Care Centres: assessment by a novel multi-approach sampling protocolPublication . Viegas, Carla; Almeida, Beatriz; Gomes, Anita Quintal; Carolino, Elisabete; Aranha Caetano, LilianaExposure to Aspergillus conidia may cause adverse effects on human health; however, no specific recommendations for routine assessments of Aspergillus in the clinical environment have been suggested so far. This study intended to determine the prevalence of Aspergillus in the clinical environment, focusing on ten Primary Health Care Centres (PHCC) through a novel multi-approach sampling protocol. Air and passive sampling, culture-based methods and a probe-based real-time assay for the detection of four clinically relevant Aspergillus sections were performed. Aspergillus spp. was observed in all PHCC, with highest prevalence on floor surface swabs (n=81) (18% on MEA; 6.94% on DG18). Regarding air samples (n=81), highest Aspergillus counts were found in the waiting room (94% MEA; 18% DG18), where Nigri was the most prevalent Aspergillus section. The use of a multi-approach sampling protocol to assess Aspergillus burden in the analyzed PHCC has greatly contributed to risk characterization, highlighting the need to implement corrective measures in order to avoid fungal presence in those settings.