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      Differences in fungi present in induced sputum samples from asthma patients and non-atopic controls: a community based case control study

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          Abstract

          Background

          There is emerging evidence for the presence of an extensive microbiota in human lungs. It is not known whether variations in the prevalence of species of microbiota in the lungs may have aetiological significance in respiratory conditions such as asthma. The aim of the study was to undertake semi-quantitative analysis of the differences in fungal species in pooled sputum samples from asthma patients and controls.

          Methods

          Induced sputum samples were collected in a case control study of asthma patients and control subjects drawn from the community in Wandsworth, London. Samples from both groups were pooled and then tested for eukaryotes. DNA was amplified using standard PCR techniques, followed by pyrosequencing and comparison of reads to databases of known sequences to determine in a semi-quantitative way the percentage of DNA from known species in each of the two pooled samples.

          Results

          A total of 136 fungal species were identified in the induced sputum samples, with 90 species more common in asthma patients and 46 species more common in control subjects. Psathyrella candolleana, Malassezia pachydermatis, Termitomyces clypeatus and Grifola sordulenta showed a higher percentage of reads in the sputum of asthma patients and Eremothecium sinecaudum, Systenostrema alba, Cladosporium cladosporioides and Vanderwaltozyma polyspora showed a higher percentage of reads in the sputum of control subjects. A statistically significant difference in the pattern of fungi that were present in the respective samples was demonstrated using the Phylogenetic (P) test (P < 0.0001).

          Conclusion

          This study is novel in providing evidence for the widespread nature of fungi in the sputum of healthy and asthmatic individuals. Differences in the pattern of fungi present in asthma patients and controls merit further investigation. Of particular interest was the presence of Malassezia pachydermatis, which is known to be associated with atopic dermatitis.

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          Most cited references11

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          Phylogenetic approaches for describing and comparing the diversity of microbial communities.

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            Analysis of fungal flora in indoor dust by ribosomal DNA sequence analysis, quantitative PCR, and culture.

            In recent years increasing attention has been given to the potential health effects of fungal exposure in indoor environments. We used large-scale sequencing of the fungal internal transcribed spacer region (ITS) of nuclear ribosomal DNA to describe the mycoflora of two office buildings over the four seasons. DNA sequencing was complemented by cultivation, ergosterol determination, and quantitative PCR analyses. Sequences of 1,339 clones were clustered into 394 nonredundant fungal operational taxonomical units containing sequences from 18 fungal subclasses. The observed flora differed markedly from that recovered by cultivation, the major differences being the near absence of several typical indoor mold genera such as Penicillium and Aspergillus spp. and a high prevalence of basidiomycetes in clone libraries. A total of 55% of the total diversity constituted of unidentifiable ITS sequences, some of which may represent novel fungal species. Dominant species were Cladosporium cladosporioides and C. herbarum, Cryptococcus victoriae, Leptosphaerulina americana and L. chartarum, Aureobasidium pullulans, Thekopsora areolata, Phaeococcomyces nigricans, Macrophoma sp., and several Malassezia species. Seasonal differences were observed for community composition, with ascomycetous molds and basidiomycetous yeasts predominating in the winter and spring and Agaricomycetidae basidiomycetes predominating in the fall. The comparison of methods suggested that the cloning, cultivation, and quantitative PCR methods complemented each other, generating a more comprehensive picture of fungal flora than any of the methods would give alone. The current restrictions of the methods are discussed.
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              Interactions of airway epithelium with protease allergens in the allergic response.

              Among the apparently innocuous environmental proteins routinely inhaled by human subjects, only a small proportion of these antigens triggers allergy in susceptible individuals. Although the molecular basis of the allergenicity of these airborne proteins remains to be fully characterized, numerous studies suggest that the ability of such proteins to promote allergic responses is at least due to their proteolytic activity. This review will summarize insights into the interactions of protease allergens with the respiratory epithelium. In addition to their capacity to facilitate their antigen presentation through epithelial barrier degradation, protease allergens can directly activate airway mucosal surfaces to recruit inflammatory cells and to initiate the airway remodelling process. A greater understanding of the effects of protease allergens in the airways inflammation as well as on the relevant targets could define novel therapeutic strategies for the treatment allergic asthma. © 2010 Blackwell Publishing Ltd.
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                Author and article information

                Contributors
                Journal
                BMC Infect Dis
                BMC Infect. Dis
                BMC Infectious Diseases
                BioMed Central
                1471-2334
                2013
                5 February 2013
                : 13
                : 69
                Affiliations
                [1 ]Institute of Primary Care & Public Health, Cardiff University School of Medicine, Neuadd Meirionnydd, Heath Park, Cardiff CF14 4YS, UK
                [2 ]Cardiff School of Bioscience, Main Building, Museum Avenue, Cardiff, CF10 3AT, UK
                Article
                1471-2334-13-69
                10.1186/1471-2334-13-69
                3570489
                23384395
                f322f692-428d-48cd-9b7e-bb9f30037391
                Copyright ©2013 van Woerden et al.; licensee BioMed Central Ltd.

                This is an Open Access article distributed under the terms of the Creative Commons Attribution License ( http://creativecommons.org/licenses/by/2.0), which permits unrestricted use, distribution, and reproduction in any medium, provided the original work is properly cited.

                History
                : 10 August 2012
                : 28 January 2013
                Categories
                Research Article

                Infectious disease & Microbiology
                asthma,sputum,fungi,case–control study
                Infectious disease & Microbiology
                asthma, sputum, fungi, case–control study

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