Portuguese cork industry: filling the knowledge gap regarding occupational exposure to fungi and related health effects.

Introduction: The presence of the Penicillium section Aspergilloides (formerly known as Penicillium glabrum) in the cork industry involves the risk of respiratory diseases such as suberosis. Methods: The aim of this study was to corroborate the predominant fungi present in this occupational environment by performing a mycological analysis of 360 workers' nasal exudates collected by nasal swabs. Additionally, evaluation of respiratory disorders among the cork workers was also performed by spirometry. Results: Penicillium section Aspergilloides was detected by qPCR in 37 out of the 360 nasal swabs collected from workers' samples. From those, 25 remained negative for Penicillium sp. when using culture-based methods. A significant association was found between ventilatory defects and years of work in the cork industry, with those people working for 10 or more years in this industry having an approximately two-fold increased risk of having ventilatory defects compared to those working less time in this setting. Among the workers who detected the presence of Penicillium section Aspergilloides, those with symptoms presented slightly higher average values of CFU. Discussion: Overall, the results obtained in this study show that working in the cork industry may have adverse effects on worker's respiratory health. Nevertheless, more studies are needed (e.g., using serological assays) to clarify the impact of each risk factor (fungi and dust) on disease etiology.

Implementation of Novel Affinity Ligand for Lentiviral Vector Purification.

The use of viral vectors as therapeutic products for multiple applications such as vaccines, cancer treatment, or gene therapies, has been growing exponentially. Therefore, improved manufacturing processes are needed to cope with the high number of functional particles required for clinical trials and, eventually, commercialization. Affinity chromatography (AC) can be used to simplify purification processes and generate clinical-grade products with high titer and purity. However, one of the major challenges in the purification of Lentiviral vectors (LVs) using AC is to combine a highly specific ligand with a gentle elution condition assuring the preservation of vector biological activity. In this work, we report for the first time the implementation of an AC resin to specifically purify VSV-G pseudotyped LVs. After ligand screening, different critical process parameters were assessed and optimized. A dynamic capacity of 1 × 1011 total particles per mL of resin was determined and an average recovery yield of 45% was found for the small-scale purification process. The established AC robustness was confirmed by the performance of an intermediate scale providing an infectious particles yield of 54%, which demonstrates the scalability and reproducibility of the AC matrix. Overall, this work contributes to increasing downstream process efficiency by delivering a purification technology that enables high purity, scalability, and process intensification in a single step, contributing to time-to-market reduction.

Assessing Multi-Attribute Characterization of Enveloped and Non-Enveloped Viral Particles by Capillary Electrophoresis.

Virus-based biopharmaceutical products are used in clinical applications such as vaccines, gene therapy, and immunotherapy. However, their manufacturing remains a challenge, hampered by the lack of appropriate analytical tools for purification monitoring or characterization of the final product. This paper describes the implementation of a highly sensitive method, capillary electrophoresis (CE)-sodium dodecyl sulfate (SDS) combined with a laser-induced fluorescence (LIF) detector to monitor the impact of various bioprocess steps on the quality of different viral vectors. The fluorescence labelling procedure uses the (3-(2-furoyl) quinoline-2-carboxaldehyde dye, and the CE-SDS LIF method enables the evaluation of in-process besides final product samples. This method outperforms other analytical methods, such as SDS-polyacrylamide gel electrophoresis with Sypro Ruby staining, in terms of sensitivity, resolution, and high-throughput capability. Notably, this CE-SDS LIF method was also successfully implemented to characterize enveloped viruses such as Maraba virus and lentivirus, whose development as biopharmaceuticals is now restricted by the lack of suitable analytical tools. This method was also qualified for quantification of rAAV2 according to the International Council for Harmonisation guidelines. Overall, our work shows that CE-SDS LIF is a precise and sensitive analytical platform for in-process sample analysis and quantification of different virus-based targets, with a great potential for application in biomanufacturing.

Nanoencapsulation of Gla-Rich Protein (GRP) as a Novel Approach to Target Inflammation.

Chronic inflammation is a major driver of chronic inflammatory diseases (CIDs), with a tremendous impact worldwide. Besides its function as a pathological calcification inhibitor, vitamin K-dependent protein Gla-rich protein (GRP) was shown to act as an anti-inflammatory agent independently of its gamma-carboxylation status. Although GRP's therapeutic potential has been highlighted, its low solubility at physiological pH still constitutes a major challenge for its biomedical application. In this work, we produced fluorescein-labeled chitosan-tripolyphosphate nanoparticles containing non-carboxylated GRP (ucGRP) (FCNG) via ionotropic gelation, increasing its bioavailability, stability, and anti-inflammatory potential. The results indicate the nanosized nature of FCNG with PDI and a zeta potential suitable for biomedical applications. FCNG's anti-inflammatory activity was studied in macrophage-differentiated THP1 cells, and in primary vascular smooth muscle cells and chondrocytes, inflamed with LPS, TNFα and IL-1ß, respectively. In all these in vitro human cell systems, FCNG treatments resulted in increased intra and extracellular GRP levels, and decreased pro-inflammatory responses of target cells, by decreasing pro-inflammatory cytokines and inflammation mediators. These results suggest the retained anti-inflammatory bioactivity of ucGRP in FCNG, strengthening the potential use of ucGRP as an anti-inflammatory agent with a wide spectrum of application, and opening up perspectives for its therapeutic application in CIDs.

Source apportionment of children daily exposure to particulate matter.

The present study aims to investigate the sources of particulate pollution in indoor and outdoor environments, with focus on determining their contribution to the exposure of children to airborne particulate matter (PM). To this end, parallel indoor and outdoor measurements were carried out for a selection of 40 homes and 5 schools between September 2017 and October 2018. PM2.5 and PM2.5-10 samples were collected during five days in each microenvironment (ME) and analysed by X-Ray Fluorescence (XRF), for the determination of elements, and by a thermal-optical technique, for the measurement of organic and elemental carbon. The source apportionment analysis of the PM composition data, by means of the receptor model SoFi (Source Finder) 8 Pro, resulted in the identification of nine sources: exhaust and non-exhaust emissions from traffic, secondary particles, heavy oil combustion, industry, sea salt, soil, city dust, and an indoor source characterized by high levels of organic carbon. Integrated daily exposure to PM2.5 was on average 21 µg/m3. The organic matter, resulting from cleaning, cooking, smoking and biological material, was the major source contributing by 31% to the PM2.5 exposure. The source city dust, which was highly influenced by the resuspension of dust in classrooms, was the second main source (26%), followed by traffic (24%). The major sources affecting the integrated exposure to PM10, which was on average 33 µg/m3, were the city dust (39%), indoor organics (24%) and traffic (16%). This study provides important information for the design of measures to reduce the exposure of children to PM.

Analysis of spatial factors, time-activity and infiltration on outdoor generated PM2.5 exposures of school children in five European cities.

Atmospheric particles are a major environmental health risk. Assessments of air pollution related health burden are often based on outdoor concentrations estimated at residential locations, ignoring spatial mobility, time-activity patterns, and indoor exposures. The aim of this work is to quantify impacts of these factors on outdoor-originated fine particle exposures of school children. We apply nested WRF-CAMx modelling of PM2.5 concentrations, gridded population, and school location data. Infiltration and enrichment factors were collected and applied to Athens, Kuopio, Lisbon, Porto, and Treviso. Exposures of school children were calculated for residential and school outdoor and indoor, other indoor, and traffic microenvironments. Combined with time-activity patterns six exposure models were created. Model complexity was increased incrementally starting from residential and school outdoor exposures. Even though levels in traffic and outdoors were considerably higher, 80-84% of the exposure to outdoor particles occurred in indoor environments. The simplest and also commonly used approach of using residential outdoor concentrations as population exposure descriptor (model 1), led on average to 26% higher estimates (15.7 µg/m3) compared with the most complex model (# 6) including home and school outdoor and indoor, other indoor and traffic microenvironments (12.5 µg/m3). These results emphasize the importance of including spatial mobility, time-activity and infiltration to reduce bias in exposure estimates.

Bioburden in sleeping environments from Portuguese dwellings.

A wider characterization of indoor air quality during sleep is still lacking in the literature. This study intends to assess bioburden before and after sleeping periods in Portuguese dwellings through active methods (air sampling) coupled with passive methods, such as electrostatic dust cloths (EDC); and investigate associations between before and after sleeping and bioburden. In addition, and driven by the lack of information regarding fungi azole-resistance in Portuguese dwellings, a screening with supplemented media was also performed. The most prevalent genera of airborne bacteria identified in the indoor air of the bedrooms were Micrococcus (41%), Staphylococcus (15%) and Neisseria (9%). The major indoor bacterial species isolated in all ten studied bedrooms were Micrococcus luteus (30%), Staphylococcus aureus (13%) and Micrococcus varians (11%). Our results highlight that our bodies are the source of the majority of the bacteria found in the indoor air of our homes. Regarding air fungal contamination, Chrysosporium spp. presented the highest prevalence both in after the sleeping period (40.8%) and before the sleeping period (28.8%) followed by Penicillium spp. (23.47% morning; 23.6% night) and Chrysonilia spp. (12.4% morning; 20.3% night). Several Aspergillus sections were identified in air and EDC samples. However, none of the fungal species/strains (Aspergillus sections Fumigati, Flavi, Nidulantes and Circumdati) were amplified by qPCR in the analyzed EDC. The correlations observed suggest reduced susceptibility to antifungal drugs of some fungal species found in sleeping environments. Toxigenic fungal species and indicators of harmful fungal contamination were observed in sleeping environments.

Chemical characterisation of particulate matter in urban transport modes.

Traffic is a main source of air pollutants in urban areas and consequently daily peak exposures tend to occur during commuting. Personal exposure to particulate matter (PM) was monitored while cycling and travelling by bus, car and metro along an assigned route in Lisbon (Portugal), focusing on PM2.5 and PM10 (PM with aerodynamic diameter <2.5 and 10 µm, respectively) mass concentrations and their chemical composition. In vehicles, the indoor-outdoor interplay was also evaluated. The PM2.5 mean concentrations were 28 ± 5, 31 ± 9, 34 ± 9 and 38 ± 21 µg/m3 for bus, bicycle, car and metro modes, respectively. Black carbon concentrations when travelling by car were 1.4 to 2.0 times higher than in the other transport modes due to the closer proximity to exhaust emissions. There are marked differences in PM chemical composition depending on transport mode. In particular, Fe was the most abundant component of metro PM, derived from abrasion of rail-wheel-brake interfaces. Enhanced concentrations of Zn and Cu in cars and buses were related with brake and tyre wear particles, which can penetrate into the vehicles. In the motorised transport modes, Fe, Zn, Cu, Ni and K were correlated, evidencing their common traffic-related source. On average, the highest inhaled dose of PM2.5 was observed while cycling (55 µg), and the lowest in car travels (17 µg). Cyclists inhaled higher doses of PM2.5 due to both higher inhalation rates and longer journey times, with a clear enrichment in mineral elements. The presented results evidence the importance of considering the transport mode in exposure assessment studies.

Advances in Lentivirus Purification.

Lentiviral vectors (LVs) have been increasingly used as a tool for gene and cell therapies since they can stably integrate the genome in dividing and nondividing cells. LV production and purification processes have evolved substantially over the last decades. However, the increasing demands for higher quantities with more restrictive purity requirements are stimulating the development of novel materials and strategies to supply the market with LV in a cost-effective manner. A detailed review of each downstream process unit operation is performed, limitations, strengths, and potential outcomes being covered. Currently, the majority of large-scale LV manufacturing processes are still based on adherent cell culture, although it is known that the industry is migrating fast to suspension cultures. Regarding the purification strategy, it consists of batch chromatography and membrane technology. Nevertheless, new solutions are being created to improve the current production schemes and expand its clinical use.

A new Hyphessobrycon (Characiformes: Characidae) of the Hyphessobrycon heterorhabdus species-group from the lower Amazon basin, Brazil

A new species of Hyphessobrycon belonging to the Hyphessobrycon heterorhabdus species-group from the lower rio Tapajós, state of Pará, Brazil, is described. The new species is allocated into the Hyphessobrycon heterorhabdus species-group due to its color pattern, composed by an anteriorly well-defined, horizontally elongated humeral blotch that becomes diffuse and blurred posteriorly, where it overlaps with a conspicuous midlateral dark stripe that becomes blurred towards the caudal peduncle and the presence, in living specimens, of a tricolored longitudinal pattern composed by a dorsal red or reddish longitudinal stripe, a middle iridescent, golden or silvery longitudinal stripe, and a more ventrally-lying longitudinal dark pattern composed by the humeral blotch and dark midlateral stripe. It can be distinguished from all other species of the group by possessing humeral blotch with a straight or slightly rounded ventral profile, lacking a ventral expansion present in all other species of the group. The new species is also distinguished from Hyphessobrycon heterorhabdus by a 9.6% genetic distance in the cytochrome c oxidase I gene. The little morphological distinction of the new species when compared with its most similar congener, H. heterorhabdus, indicates that the new species is one of the first truly cryptic fish species described from the Amazon basin.(AU)

Uma nova espécie de Hyphessobrycon pertencente ao grupo Hyphessobrycon heterorhabdus é descrita da região do baixo rio Tapajós, estado do Pará, Brasil. A nova espécie é incluída no grupo Hyphessobrycon heterorhabdus devido ao seu padrão de coloração, composto por uma mancha umeral alongada, anteriormente bem definida, que se torna difusa e borrada posteriormente, onde se sobrepõe a uma conspícua faixa escura médio-lateral que se torna borrada próxima ao pedúnculo caudal, e pela presença, em exemplares vivos, de um padrão longitudinal tricolor, composto por uma faixa longitudinal vermelha ou avermelhada dorsal, uma faixa média iridescente dourada ou prateada e, mais ventralmente, o padrão longitudinal escuro composto pela faixa escura médio-lateral e mancha umeral. A espécie pode ser distinguida das outras espécies pertencentes ao grupo por possuir uma mancha umeral com região ventral retilínea ou levemente arredondada, sem uma expansão ventral presente nas demais espécies do grupo. A espécie também se diferencia de Hyphessobrycon heterorhabdus por uma distância genética de 9,6% no gene citocromo c oxidase I. A sutil diferença morfológica da nova espécie quando comparada ao seu congênere mais similar, H. heterorhabdus, indica que a nova espécie é uma das primeiras espécies de peixes verdadeiramente crípticas descritas da Bacia Amazônica.(AU)

Mercury Removal from Aqueous Solution Using ETS-4 in the Presence of Cations of Distinct Sizes.

The removal of the hazardous Hg2+ from aqueous solutions was studied by ion exchange using titanosilicate in sodium form (Na-ETS-4). Isothermal batch experiments at fixed pH were performed to measure equilibrium and kinetic data, considering two very distinct situations to assess the influence of competition effects: (i) the counter ions initially in solution are Na+ and Hg2+ (both are exchangeable); (ii) the initial counter ions in solution are tetrapropylammonium (TPA+) and Hg2+ (only Hg2+ is exchangeable, since TPA+ is larger than the ETS-4 micropores). The results confirmed that ETS-4 is highly selective for Hg2+, with more than 90% of the mercury being exchanged from the fluid phase. The final equilibrium attained under the presence of TPA+ or Na+ in solution was very similar, however, the Hg2+/Na+/ETS-4 system in the presence of Na+ required more 100 h to reach equilibrium than in the presence of TPA+. The Hg2+/Na+/ETS-4 system was modelled and analyzed in terms of equilibrium (mass action law) and mass transfer (Maxwell-Stefan (MS) formalism). Concerning equilibrium, no major deviations from ideality were found in the range of studied concentrations. On the other hand, the MS based model described successfully (average deviation of 5.81%) all kinetic curves of mercury removal.

A new Hyphessobrycon (Characiformes: Characidae) of the Hyphessobrycon heterorhabdus species-group from the Central Amazon basin, Brazil.

A new species of Hyphessobrycon Durbin from the Paraná do Urariá system in Central Amazon region, Amazonas state, Brazil, is described. The new species is allocated into the Hyphessobrycon heterorhabdus species-group due to its color pattern, composed by a well-defined, horizontally elongated humeral blotch continuous with a conspicuous midlateral dark stripe that becomes blurred towards the caudal peduncle, and can be distinguished from all other species of the group by possessing humeral blotch and continuous midlateral stripe broad, occupying vertical height equivalent of two scale rows. A tricolored pattern composed dorsally by a red or reddish longitudinal stripe, a middle iridescent, golden or silvery longitudinal stripe, and ventrally by a variably-developed longitudinal dark stripe is identified as a putative additional character shared by the species of the Hyphessobrycon heterorhabdus species-group. The presence of bony hooks in all fins in mature males of some species of the Hyphessobrycon heterorhabdus species-group is also discussed.

Compliance of indoor air quality during sleep with legislation and guidelines - A case study of Lisbon dwellings.

This study aimed to provide a comprehensive characterisation of the indoor air quality during the sleeping period of 10 couples at Lisbon dwellings, using a multi-pollutant approach, and to understand how the compliance with legislation and guidelines was to assure a good indoor air quality. The assessment of indoor air quality was conducted in the cold season using real time monitors during the sleeping period for comfort parameters (temperature and relative humidity) and air pollutants (carbon dioxide - CO2, carbon monoxide - CO, formaldehyde - CH2O, total volatile organic compounds - VOCs, and particulate matter - PM2.5 and PM10), together with active sampling of bioaerosols (fungi and bacteria) before and after the sleeping period. Lower compliance (less than 50% of the cases) with the Portuguese legislation was found for temperature, CO2 (3440 ± 1610 mg m-3), VOCs (1.79 ± 0.99 mg m-3) and both bioaerosol types. In 70% of the cases, PM2.5 (15.3 ± 9.1 µg m-3) exceeded the WHO guideline of 10 µg m-3. All bedrooms presented air change rates above the recommended minimum value of 0.7 h-1, highlighting that a good indoor air quality during sleep is not guaranteed.

Relationship between indoor and outdoor size-fractionated particulate matter in urban microenvironments: Levels, chemical composition and sources.

Exposure to particulate matter (PM) has been associated with adverse health outcomes, particularly in susceptible population groups such as children. This study aims to characterise children's exposure to PM and its chemical constituents. Size-segregated aerosol samples (PM0.25, PM0.25-0.5, PM0.5-1.0, PM1.0-2.5 and PM2.5-10) were collected in the indoor and outdoor of homes and schools located in Lisbon (Portugal). Organic and elemental carbon (OC and EC) were determined by a thermo-optical method, whereas major and trace elements were analysed by X-Ray Fluorescence. In school, the children were exposed to higher PM concentrations than in home, which might be associated not only to the elevated human occupancy but also to outdoor infiltration. The pattern of PM mass size distribution was dependent on the location (home vs. school and indoor vs. outdoor). The presence of EC in PM0.25 and OC in PM0.25-0.5 was linked to traffic exhaust emissions. OC and EC in PM2.5-10 may be explained by their adhesion to the surface of coarser particles. Generally, the concentrations of mineral and marine elements increased with increasing PM size, while for anthropogenic elements happened the opposite. In schools, the concentrations of mineral matter, anthropogenic elements and marine aerosol were higher than in homes. High mineral matter concentrations found in schools were related to the close proximity to busy roads and elevated human occupancy. Overall, the results suggest that exposure to PM is relevant and highlights the need for strategies that provide healthier indoor environments, principally in schools.

Characterization of Occupational Exposure To Fungal Burden in Portuguese Bakeries.

Several 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.

Antifungal-resistant Mucorales in different indoor environments.

This paper sought to address the prevalence of Mucorales in different indoor environments in Portugal. Environmental samples (183 in total) were collected at dwellings (n = 79) and workplaces (bakeries, swine farms, taxis, waste-sorting plants) (n = 93) by passive sampling using electrostatic dust collector (EDC), air-conditioning filters, litter, and/or raw materials. Samples were inoculated onto non-selective MEA and DG18 media and were screened for antifungal drug-resistance in azole-supplemented agar Sabouraud media. A probe-based Mucorales-specific real-time PCR assay (Muc18S) was used to detect Mucorales in complement to conventional culture-based methods. Mucorales order was found as more prevalent in air-conditioning filters from waste-sorting fork lifters (35.7%). Amongst Mucorales isolates able to grow in azole-supplemented media, 16 isolates of Mucor sp., Rhizopus sp. or Rhizomucor sp. were not susceptible to 1 mg/L voriconazole, and four isolates of Mucor sp. or Rhizopus sp. were not susceptible to 4 mg/L itraconazole. In conclusion, combination of the culture-based and molecular methods proved to be reliable for Mucorales order identification in complex environmental samples.

Fungal diversity and mycotoxin distribution in echinoderm aquaculture.

Aquaculture has been a growing sector of food production worldwide in the last decades, and now starts to include new, unconventional species from the Phylum Echinodermata, such as sea urchin (Paracentrotus lividus) and sea cucumber (Holothuria tubulosa). However, little is known in this context with regard to food safety aspects arising from toxigenic fungi. In this study, samples of feed (n = 7) and water (n = 8) or water filters (n = 4) from experimental aquaculture systems, producing sea urchin and sea cucumber, were analyzed by culture-based microbiological methods to assess fungal associations. Additionally, a search using molecular techniques for toxigenic sections within the genus Aspergillus in these materials was done. Finally, samples were analyzed for 37 mycotoxins by LC-MS/MS. In feed samples, Fusarium verticillioides and F. culmorum were detected. In water and water filter samples, Aureobasidium spp., Penicillium spp., and Cladosporium spp. were found. No genes of species from toxigenic Aspergillus sections were detected. Some feed samples were contaminated by multiple mycotoxins, namely deoxynivalenol (DON), zearalenone (ZEN), fumonisins (FBs), T-2 toxin, ochratoxin A (OTA), and mycophenolic acid (MPA). This is the first one study dealing with toxigenic fungi and mycotoxins in echinoderm-producting aquaculture. Although no clear evidence for adverse effects on the production systems could be found, the confirmed environmental association of mycotoxins and echinoderms requires further consideration. Studies on the consequences of introducing cereal-based fungi and their mycotoxins via feeds into aquaculture systems for echinoderm production seem to be advisable, to assess possible adverse effects on production and to clarify the potential impact on public health.

Aspergillus prevalence in air conditioning filters from vehicles: Taxis for patient transportation, forklifts, and personal vehicles.

The 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.

Filters from taxis air conditioning system: A tool to characterize driver's occupational exposure to bioburden?

Bioburden proliferation in filters from air conditioning systems of taxis represents a possible source of occupational exposure. The aim of this study was to determine the occurrence of fungi and bacteria in filters from the air conditioning system of taxis used for patient transportation and to assess the exposure of drivers to bioburden. Filters from the air conditioning systems of 19 taxis and 28 personal vehicles (used as controls) operating in three Portuguese cities including the capital Lisbon, were collected during the winter season. The occurrence and significance of bioburden detected in the different vehicles are reported and discussed in terms of colony-forming units (CFU) per 1 m2 of filter area and by the identification of the most frequently detected fungal isolates based on morphology. Azole-resistant mycobiota, fungal biomass, and molecular detection of Aspergillus species/strains were also determined. Bacterial growth was more prevalent in taxis (63.2%) than in personal vehicles (26.3%), whereas fungal growth was more prevalent in personal vehicles (53.6%) than in taxis (21.1-31.6%). Seven different azole-resistant species were identified in this study in 42.1% taxi filters. Levels of fungal biomass were above the detection limit in 63% taxi filters and in 75% personal vehicle filters. No toxigenic species were detected by molecular analysis in the assessed filters. The results obtained show that bioburden proliferation occurs widely in filters from the air conditioning systems of taxis, including the proliferation of azole-resistant fungal species, suggesting that filters should be replaced more frequently. The use of culture based-methods and molecular tools combined enabled an improved risk characterization in this setting.

Fungal burden exposure assessment in podiatry clinics from Ireland.

Fungi are amongst the bioaerosols of most importance, as indicated by the growing interest in this field of research. The aim was to characterize the exposure to fungal burden in podiatry clinics using culture-based and molecular methods. METHODS: Airborne fungi were collected using an impaction air sampler and surface samples were also performed. Fourteen air samples were collected for direct detection of fungal DNA from filamentous fungi and dermatophytes. Overall, 63.6 % of the evening samples and 46 % of the morning samples surpassed the threshold values (150 CFU/m3). Molecular detection, by real time PCR, of the target fungal species/strains (Aspergillus and Stachybotrys species) was negative for all samples collected. Trichophyton rubrum was detected by PCR analysis in one DNA sample collected on day six. Results suggest the use of both culture-based and molecular methodologies are desirable for a complete evaluation of fungal burden in this particular health care setting.