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DNA analysis of outdoor air reveals a high degree of fungal diversity, temporal variability, and genera not seen by spore morphology. Fungal Biology, 116, 214-224
Institute for Lung Health, Department of Infection, Immunity and Inflammation, University of Leicester, Leicester LE1 9HN, UK.
Fungal Biology (Impact Factor: 2.34). 02/2012; 116(2):214-24. DOI: 10.1016/j.funbio.2011.11.004 Source: PubMed
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Available from: Catherine H. Pashley, Jan 06, 2014 Data provided are for informational purposes only. Although carefully collected, accuracy cannot be guaranteed. The impact factor represents a rough estimation of the journal's impact factor and does not reflect the actual current impact factor. Publisher conditions are provided by RoMEO. Differing provisions from the publisher's actual policy or licence agreement may be applicable.
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- "Recently, it has been found that the fungal aerosol may affect atmospheric chemistry and may act as cloud condensation nuclei influencing the Earth's climate (Després et al. 2012). Recent studies with the implementation of molecular techniques demonstrated that the fungal spectrum suspended in the air is much richer than previously known (Fröhlich- Nowoisky et al. 2009; Pashley et al. 2012). Although the fungal spore content of the atmosphere has attracted significant interest during recent decades, it still remains underexplored in many parts of the world. "
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ABSTRACT: The occurrence and the abundance of the airborne fungi in the city of Athens have been studied by a volumetric non-culture-based method. During a period of 4 years, 562 glass slides were exposed during 562 sampling days, by a Burkard portable air sampler. A total of 259,851 fungal spores have been recovered and assigned to 24 genera of filamentous fungi, in addition to those recorded as groups. The annual mean concentration (AMC) of the total fungi was 865, 1,212, 937 and 1,206 spores/m 3 , and the concentration range was 89–3,175, 63–7,628, 62–4,323 and 29–10,187 spores/m 3 for each year, respectively. There is no statistically significant variation in the AMC and in the distribution patterns of the total fungi from year to year. The temporal variation of the total fungi and of the dominant genera Cladosporium and Alternaria presented a significant increase during the warm months of each year. The results of the present study compared with data available by a culture-based method confirmed the complementarity of the two sampling procedures. The non-culture-based method was more efficient for the quantification of the total fungal count as well as of the genera Cladosporium and Alternaria, although less effective for the quantification of the genera Asper-gillus and Penicillium. -
- "These were subjected to 2 min bead-bashing (BioSpec mini bead beater-16) then incubated for 10 min at 65 °C. Total genomic DNA was extracted using the DNeasy plant mini kit (Qiagen) following the procedure reported in [14]. "
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ABSTRACT: Next generation sequencing technology has revolutionised microbiology by allowing concurrent analysis of whole microbial communities. Here we developed and verified similar methods for the analysis of fungal communities using a proton release sequencing platform with the ability to sequence reads of up to 400 bp in length at significant depth. This read length permits the sequencing of amplicons from commonly used fungal identification regions and thereby taxonomic classification. Using the 400 bp sequencing capability, we have sequenced amplicons from the ITS1, ITS2 and LSU fungal regions to a depth of approximately 700,000 raw reads per sample. Representative operational taxonomic units (OTUs) were chosen by the USEARCH algorithm, and identified taxonomically through nucleotide blast (BLASTn). Combination of this sequencing technology with the bioinformatics pipeline allowed species recognition in two controlled fungal spore populations containing members of known identity and concentration. Each species included within the two controlled populations was found to correspond to a representative OTU, and these OTUs were found to be highly accurate representations of true biological sequences. However, the absolute number of reads attributed to each OTU differed among species. The majority of species were represented by an OTU derived from all three genomic regions although in some cases, species were only represented in two of the regions due to the absence of conserved primer binding sites or due to sequence composition. It is apparent from our data that proton release sequencing technologies can deliver a qualitative assessment of the fungal members comprising a sample. The fact that some fungi cannot be amplified by specific "conserved" primer pairs confirms our recommendation that a multi-region approach be taken for other amplicon-based metagenomic studies. -
- "Generally molecular methods for fungal identification that employ diversity of these regions rely on amplification of the sample, followed by either Sanger-clone based sequencing, hybridisation to specific probes or HTS methods to analyse the regions. Sangersequencing strategies have been successfully used by Pashley et al. (2012) and others (Robertson et al., 2013) for accurate fungal identification. However, this method is laborious, requiring the picking of multiple clones followed by ligation of the amplification product into a suitable vector, making it unsuitable for routine monitoring. "
Article: Workshop on the sources, quantification and health implications of bioaerosols workshop report
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ABSTRACT: Scientists, health care professionals and the general public have all raised concerns about the potential health impact natural and manmade sources bioaerosols. In response in september 2012 the Natural Environmental Research Council and public health England jointly funded a workshop to review the 'sources, quantification and health implications of bioaerosols'. The organising committee for the workshop identified five priority areas-(1) bioaerosol identification and quantification methodology, (2) bioaerosol sources, (3) health effects of bioaerosols, (4) extreme events, risk assessment and mediation and (5) bioaerosol dispersion and modelling. The primary purpose was to bring together experts to report on recent research and identify research gaps where increased knowledge would improve risk understanding and public health. This report summarises the presentations, the main discussion points and key conclusions that emerged during the workshop.
