Browsing by Author "Viegas, S."
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- Accessing indoor fungal contamination by Aspergillus fumigatus complex using conventional and molecular methods in portuguese poultriesPublication . Viegas, C.; Malta-Vacas, J.; Sabino, R.; Viegas, S.; Veríssimo, C.Epidemiological studies showed increased prevalence of respiratory symptoms and adverse changes in pulmonary function parameters in poultry workers, corroborating the increased exposure to risk factors, such as fungal load and their metabolites. This study aimed to determine the occupational exposure threat due to fungal contamination caused by the toxigenic isolates belonging to the complex of the species of Aspergillus flavus and also isolates from Aspergillus fumigatus species complex. The study was carried out in seven Portuguese poultries, using cultural and molecular methodologies. For conventional/cultural methods, air, surfaces, and litter samples were collected by impaction method using the Millipore Air Sampler. For the molecular analysis, air samples were collected by impinger method using the Coriolis μ air sampler. After DNA extraction, samples were analyzed by real-time PCR using specific primers and probes for toxigenic strains of the Aspergillus flavus complex and for detection of isolates from Aspergillus fumigatus complex. Through conventional methods, and among the Aspergillus genus, different prevalences were detected regarding the presence of Aspergillus flavus and Aspergillus fumigatus species complexes, namely: 74.5 versus 1.0 % in the air samples, 24.0 versus 16.0 % in the surfaces, 0 versus 32.6 % in new litter, and 9.9 versus 15.9 % in used litter. Through molecular biology, we were able to detect the presence of aflatoxigenic strains in pavilions in which Aspergillus flavus did not grow in culture. Aspergillus fumigatus was only found in one indoor air sample by conventional methods. Using molecular methodologies, however, Aspergillus fumigatus complex was detected in seven indoor samples from three different poultry units. The characterization of fungal contamination caused by Aspergillus flavus and Aspergillus fumigatus raises the concern of occupational threat not only due to the detected fungal load but also because of the toxigenic potential of these species.
- Accessing occupational exposure to fungi in a cork industryPublication . Viegas, C.; Clérigo, A.; Faria, T.; Sabino, Raquel; Veríssimo, Cristina; Quintal-Gomes, A.; Viegas, S.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. Nasal mucous samples from 127 workers were taken with sterilized cotton swabs.The fungal species identified in the collected nose swabs were shown to be correlated with the results obtained in the environment. Eighty workers (63.0%) presented contamination of their nose nostril with Chrysonilia sitophila, which number of colonies was countless. Within the Aspergillus genus, the complexes Fumigati, Circumdati, Versicolores and Candidi were isolated. No azole-resistant Aspergillus isolates grew in the selective media used (screened itraconazole and voriconazole resistance).This approach allowed us to estimate the risk associated with these tasks performance. Moreover, the cork industry is related to high dust contamination and this can promote exposure to fungi since dust particles can act as carriers of fungi to the worker’s nose. Assessment by molecular tools will ensure the specific targeting of DNA from P. glabrum complex in workers nose.
- Accuracy of prenatal culture in predicting intrapartum group B streptococcus colonization statusPublication . Florindo, C.; Damião, V.; Lima, J.; Nogueira, I.; Rocha, I.; Caetano, P.; Ribeiro, L.; Viegas, S.; Gomes, João Paulo; Borrego, M.J.Objective: To evaluate the positive predictive value (PPV) of group B Streptococcus (GBS) cultures at 35–37 weeks of gestation relative to GBS colonization status at delivery. Methods: Rectovaginal swabs from 221 women at labor in four Lisbon hospitals were collected for GBS screening according to the CDC guidelines. Results: The PPV was 24.4%. IAP was administered to 100% of prenatally GBS positive women. There was no case of early onset GBS disease (EOD). Conclusions: Poor accuracy of prenatal cultures in identifying true candidates for IAP highlights the need for Portuguese clinical and laboratory guidelines to prevent EOD and antibiotic overtreatment of pregnant women.
- Air contaminants in animal production – poultry casePublication . Viegas, C.; Viegas, S.; Monteiro, A.; Carolino, E.; Sabino, R.; Verissimo, C.A descriptive study was developed in order to assess air contamination caused by fungi and particles in seven poultry units. Twenty seven air samples of 25 litters were collected through impaction method. Air sampling and particle concentration measurement were performed in the pavilions’ interior and also outside premises, since this was the place regarded as reference. Simultaneously, temperature and relative humidity were also registered. Regarding fungal load in the air from the seven poultry farms, the highest value obtained was 24040 CFU/m3 and the lowest was 320 CFU/m3. Twenty eight species/genera of fungi were identified, being Scopulariopsis brevicaulis (39.0%) the most commonly isolated species and Rhizopus sp. (30.0%) the most commonly isolated genus. From the Aspergillus genus, Aspergillus flavus (74.5%) was the most frequently detected species. There was a significant correlation (r=0.487; p=0.014) between temperature and the level of fungal contamination (CFU/m3). Considering contamination caused by particles, in this study, particles with larger dimensions (PM5.0 and PM10) have higher concentrations. There was also a significant correlation between relative humidity and concentration of smaller particles namely, PM0.5 (r=0.438; p=0.025) and PM1.0 (r=0.537; p=0.005). Characterizing typical exposure levels to these contaminants in this specific occupational setting is required to allow a more detailed risk assessment analysis and to set exposure limits to protect workers’ health.
- Aspergillus flavus contamination in 2 Portuguese waste water treatment plants: an occupational threatPublication . Viegas, C.; Dias, R.; Gomes, A.Q.; Meneses, M.; Sabino, R.; Viegas, S.Filamentous fungi from genus Aspergillus were previously detected in wastewater treatment plants (WWTP) as being Aspergillus flavus (A. flavus), an important toxigenic fungus producing aflatoxins. This study aimed to determine occupational exposure adverse effects due to fungal contamination produced by A. flavus complex in two Portuguese WWTP using conventional and molecular methodologies. Air samples from two WWTP were collected at 1 m height through impaction method. Surface samples were collected by swabbing surfaces of the same indoor sites. After counting A. flavus and identification, detection of aflatoxin production was ensured through inoculation of seven inoculates in coconut-milk agar. Plates were examined under long-wave ultraviolet (UV; 365 nm) illumination to search for the presence of fluorescence in the growing colonies. To apply molecular methods, air samples were also collected using the impinger method. Samples were collected and collection liquid was subsequently used for DNA extraction. Molecular identification of A. flavus was achieved by real-time polymerase chain reaction (RT-PCR) using the Rotor-Gene 6000 qPCR detection system (Corbett). Among the Aspergillus genus, the species that were more abundant in air samples from both WWTP were Aspergillus versicolor (38%), Aspergillus candidus (29.1%), and Aspergillus sydowii (12.7%). However, the most commonly species found on surfaces were A. flavus (47.3%), Aspergillus fumigatus (34.4%), and Aspergillus sydowii (10.8%). Aspergillus flavus isolates that were inoculated in coconut agar medium were not identified as toxigenic strains and were not detected by RT-PCR in any of the analyzed samples from both plants. Data in this study indicate the need for monitoring fungal contamination in this setting. Although toxigenic strains were not detected from A. flavus complex, one cannot disregard the eventual presence and potential toxicity of aflatoxins.
- Assessment of fungal contamination in waste sorting and incineration-case study in PortugalPublication . Viegas, C.; Gomes, Q.; Abegão, J.; Sabino, R.; Graça, T.; Viegas, S.Organic waste is a rich substrate for microbial growth, and because of that, workers from waste industry are at higher risk of exposure to bioaerosols. This study aimed to assess fungal contamination in two plants handling solid waste management. Air samples from the two plants were collected through an impaction method. Surface samples were also collected by swabbing surfaces of the same indoor sites. All collected samples were incubated at 27◦C for 5 to 7 d. After lab processing and incubation of collected samples, quantitative and qualitative results were obtained with identification of the isolated fungal species. Air samples were also subjected to molecular methods by real-time polymerase chain reaction (RT PCR) using an impinger method to measure DNA of Aspergillus flavus complex and Stachybotrys chartarum. Assessment of particulate matter (PM) was also conducted with portable direct-reading equipment. Particles concentration measurement was performed at five different sizes (PM0.5; PM1; PM2.5; PM5; PM10). With respect to the waste sorting plant, three species more frequently isolated in air and surfaces were A. niger (73.9%; 66.1%), A. fumigatus (16%; 13.8%), and A. flavus (8.7%; 14.2%). In the incineration plant, the most prevalent species detected in air samples were Penicillium sp. (62.9%), A. fumigatus (18%), and A. flavus (6%), while the most frequently isolated in surface samples were Penicillium sp. (57.5%), A. fumigatus (22.3%) and A. niger (12.8%). Stachybotrys chartarum and other toxinogenic strains from A. flavus complex were not detected. The most common PM sizes obtained were the PM10 and PM5 (inhalable fraction). Since waste is the main internal fungal source in the analyzed settings, preventive and protective measures need to be maintained to avoid worker exposure to fungi and their metabolites.
- Climate change and aflatoxins exposure in Portugal – could we expect a significant health risk?Publication . Assunção, R.; Martins, C.; Viegas, S.; Viegas, C.; Jakobsen, Lea S.; Pires, S.; Alvito, P.In recent decades, changes in climate have caused impacts on natural and human systems on all continents and across the oceans. Climate change (CC) has become one of the most critical issues for the sustainable development of human societies and the functioning of ecosystems on earth. Portugal is highly vulnerable to CC impacts due to its Europe south-western geographical situation, mainly through decreasing annual precipitation, more intense extreme weather and climate events. In fact, CC is considered as one of the biggest global threats to human health of the 21st century and its effects will undoubtedly influence agricultural systems and food safety. Due to the potential impact on the occurrence of food hazards, increased concern has being aroused. Mycotoxins, natural contaminants produced by fungi, are among the most important of such hazards, especially due to their potential to establish deleterious health conditions. From all known mycotoxins, aflatoxins stand to the most toxic for humans. Dietary aflatoxins exposure is considered a significant risk factor for hepatocellular carcinoma (HCC). Considering the impact of CC in mycotoxin contamination of food products available in Portugal, the present study aims to discuss the potential influence of CC in the health risk associated to aflatoxins dietary exposure of Portuguese population. To tackle this objective, the estimated number of hepatocellular carcinoma (HCC) and the associated burden of disease in terms of Disability Adjusted Life Years (DALYs) were derived. Results showed that the number of extra cases of HCC associated to Portuguese aflatoxin dietary exposure ranged from 0.52 and 2.00. The derived DALYs ranged between 8.0 and 30.9. It is expected that in the future the number of DALYs and the associated HCC due to aflatoxin exposure should increase due to CC. These results should represent an alert for the potential consequences of an incompletely explored perspective of CC – the health impact of the human exposure to aflatoxins. Politics and decision-makers should be involved and committed to implement effective measures to deal with CC issues and to reduce its possible consequences.
- Comparison of indoor and outdoor fungi and particles in poultry unitsPublication . Viegas, C.; Viegas, S.; Monteiro, A.; Carolino, E.; Sabino, R.; Verissimo, C.A descriptive study was developed in order to compare indoor and outdoor air contamination caused by fungi and particles in seven poultry units. Twenty eight air samples of 25 litters were collected through the impaction method on malt extract agar. Air sampling and particles concentration measurement were done in the interior and also outside premises of the poultries’ pavilions. Regarding the fungal load in the air, indoor concentration of mold was higher than outside air in six poultry units. Twenty eight species / genera of fungi were identified indoor, being Scopulariopsis brevicaulis (40.5%) the most commonly isolated species and Rhizopus sp. (30.0%) the most commonly isolated genus. Concerning outdoor, eighteen species/genera of fungi were isolated, being Scopulariopsis brevicaulis (62.6%) also the most isolated. All the poultry farms analyzed presented indoor fungi different from the ones identified outdoors. Regarding particles’ contamination, PM2.5, PM5.0 and PM10 had a statistically significant difference (Mann-Whitney U test) between the inside and outside of the pavilions, with the inside more contaminated (p=.006; p=.005; p=.005, respectively). The analyzed poultry units are potential reservoirs of substantial amounts of fungi and particles and could therefore free them in the atmospheric air. The developed study showed that indoor air was more contaminated than outdoors, and this can result in emission of potentially pathogenic fungi and particles via aerosols from poultry units to the environment, which may post a considerable risk to public health and contribute to environmental pollution.
- Drinking Green Tea: Despite the Risks Due to Mycotoxins, Is It Possible to Increase the Associated Health Benefits?Publication . Assunção, Ricardo; Twarużek, M; Kosicki, R.; Viegas, C.; Viegas, S.Tea has been consumed for thousands of years. Despite the different varieties, particular emphasis has been placed on green tea (GT), considering the associated health benefits following its regular consumption, some of which are due to its polyphenol constituents, such as epigallocatechin- 3-gallate (EGCG). Tea is not prone to the growth of microorganisms, except fungus, when proper storage, handling, and packing conditions are compromised. Consequently, mycotoxins, secondary metabolites of fungi, could contaminate tea samples, affecting human health. In the present study, we aimed to assess the balance between risks (due to mycotoxins and high levels of EGCG) and benefits (due to moderate intake of EGCG) associated with the consumption of GT. For this, 20 GT samples (10 in bulk and 10 in bags) available in different markets in Lisbon were analyzed through a LC–MS/MS method, evaluating 38 different mycotoxins. Six samples revealed detectable values of the considered toxins. Current levels of mycotoxins and EGCG intake were not associated with health concerns. Scenarios considering an increasing consumption of GT in Portugal showed that drinking up to seven cups of GT per day should maximize the associated health benefits. The present study contributes to the future establishment of GT consumption recommendations in Portugal.
- Effect biomarkers in e-waste management workersPublication . Silva, Maria João; Aimonen, K.; Louro, Henriqueta; Tavares, A.; Moreira, R.; Catalan, J.; Duca, R.C.; Godderis, L.; Mahiout, S.; Martins, C.; Martinsone, I.; Matisane, L.; Namorado, S.; Van Nieuwenhuyse, A.; Pinhal, H.; Porras, S.; Remes, J.; Scheepers, P.; Verdonck, J.; Viegas, S.; Santonen, T.; HBM4EU E-waste study teamDuring e-waste handling/processing, a broad range of toxic chemicals (metals and persistent organic compounds), are released and may affect workers’ health. This work intended to identify genotoxic effects in workers from European e-waste management companies. Micronuclei were analysed in peripheral blood lymphocytes (MNPBL) from 95 workers and 50 controls and in reticulocytes (MNRET) from 82 workers and 41 controls. No statistically significant differences were detected between the total exposed and control groups, for both MNPBL and MNRET frequencies. Stratification of workers in subgroups according to the main activities performed revealed that the subgroup involved in batteries recycling (n=23) presented a frequency of MNPBL significantly higher than that of controls. Significant differences in MNPBL frequencies were also found between battery workers and the subgroups handling/processing white goods, metals and plastics, and miscellaneous E-waste; no differences in MNRET frequencies among subgroups were detected. Worth to note, the subgroup dealing with brown goods (n=12) displayed the highest MNPBL and MNRET frequencies, although statistical significances were not observed when comparing with the other subgroups or controls. These preliminary results highlight the value of adding effect biomarkers to biomonitoring campaigns, to uncover groups of workers at enhanced risk and to prioritize risk management measures’ implementation.