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Published in: BMC Infectious Diseases 1/2013

Open Access 01-12-2013 | Research article

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

Authors: Hugo Cornelis van Woerden, Clive Gregory, Richard Brown, Julian Roberto Marchesi, Bastiaan Hoogendoorn, Ian Price Matthews

Published in: BMC Infectious Diseases | Issue 1/2013

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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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Literature
2.
go back to reference Bartsch P, Collignon A, Weber G, Robaye G, Delbrouck JM, Roelandts I, Yujie J: Distribution of metals in human lung: analysis by particle induced x-ray emission. Arch Environ Health. 1982, 37 (2): 111-117.CrossRefPubMed Bartsch P, Collignon A, Weber G, Robaye G, Delbrouck JM, Roelandts I, Yujie J: Distribution of metals in human lung: analysis by particle induced x-ray emission. Arch Environ Health. 1982, 37 (2): 111-117.CrossRefPubMed
3.
go back to reference Bunnell J: Editorial. Epidemio-Ecology News. 2002, 1 (2): 1- Bunnell J: Editorial. Epidemio-Ecology News. 2002, 1 (2): 1-
4.
go back to reference Griffin DW, Kellogg CA, Garrison VH, Holmes C, Shinn EA: The movement of soil and sediment in Earth's atmosphere: microbiology and ecosystem health. Epidemio-Ecology News. 2002, 1 (2): 1-6. Griffin DW, Kellogg CA, Garrison VH, Holmes C, Shinn EA: The movement of soil and sediment in Earth's atmosphere: microbiology and ecosystem health. Epidemio-Ecology News. 2002, 1 (2): 1-6.
5.
go back to reference World Health Organisation: Hazard prevention and control in the work environment: airborne dust. 1999, Geneva: WHO World Health Organisation: Hazard prevention and control in the work environment: airborne dust. 1999, Geneva: WHO
6.
go back to reference Scott JAG, Hall AJ: The value and complications of percutaneous transthoracic lung aspiration for the etiologic diagnosis of community-acquired pneumonia*. Chest. 1999, 116 (6): 1716-1732. 10.1378/chest.116.6.1716.CrossRefPubMed Scott JAG, Hall AJ: The value and complications of percutaneous transthoracic lung aspiration for the etiologic diagnosis of community-acquired pneumonia*. Chest. 1999, 116 (6): 1716-1732. 10.1378/chest.116.6.1716.CrossRefPubMed
7.
go back to reference Rogers GB, Carroll MP, Serisier DJ, Hockey PM, Jones G, Kehagia V, Connett GJ, Bruce KD: Use of 16S rRNA gene profiling by terminal restriction fragment length polymorphism analysis to compare bacterial communities in sputum and mouthwash samples from patients with cystic fibrosis. J Clin Microbiol. 2006, 44 (7): 2601-2604. 10.1128/JCM.02282-05.CrossRefPubMedPubMedCentral Rogers GB, Carroll MP, Serisier DJ, Hockey PM, Jones G, Kehagia V, Connett GJ, Bruce KD: Use of 16S rRNA gene profiling by terminal restriction fragment length polymorphism analysis to compare bacterial communities in sputum and mouthwash samples from patients with cystic fibrosis. J Clin Microbiol. 2006, 44 (7): 2601-2604. 10.1128/JCM.02282-05.CrossRefPubMedPubMedCentral
8.
go back to reference Tunney MM, Field TR, Moriarty TF, Patrick S, Doering G, Muhlebach MS, Wolfgang M, Boucher R, Gilpin DF, McDowell A: Detection of anaerobic bacteria in high numbers in sputum from patients with cystic fibrosis. Am J Respir Crit Care Med. 2008, 177: 995-1001. 10.1164/rccm.200708-1151OC.CrossRefPubMed Tunney MM, Field TR, Moriarty TF, Patrick S, Doering G, Muhlebach MS, Wolfgang M, Boucher R, Gilpin DF, McDowell A: Detection of anaerobic bacteria in high numbers in sputum from patients with cystic fibrosis. Am J Respir Crit Care Med. 2008, 177: 995-1001. 10.1164/rccm.200708-1151OC.CrossRefPubMed
9.
go back to reference Rogers GB, Carroll MP, Serisier DJ, Hockey PM, Kehagia V, Jones GR, Bruce KD: Bacterial activity in cystic fibrosis lung infections. Respir Res. 2005, 6 (1): 49-10.1186/1465-9921-6-49.CrossRefPubMedPubMedCentral Rogers GB, Carroll MP, Serisier DJ, Hockey PM, Kehagia V, Jones GR, Bruce KD: Bacterial activity in cystic fibrosis lung infections. Respir Res. 2005, 6 (1): 49-10.1186/1465-9921-6-49.CrossRefPubMedPubMedCentral
10.
go back to reference Hilty M, Burke C, Pedro H, Cardenas P, Bush A, Bossley C, Davies J, Ervine A, Poulter L, Pachter L, et al: Disordered microbial communities in asthmatic airways. PLoS One. 2010, 5 (1): e8578-10.1371/journal.pone.0008578.CrossRefPubMedPubMedCentral Hilty M, Burke C, Pedro H, Cardenas P, Bush A, Bossley C, Davies J, Ervine A, Poulter L, Pachter L, et al: Disordered microbial communities in asthmatic airways. PLoS One. 2010, 5 (1): e8578-10.1371/journal.pone.0008578.CrossRefPubMedPubMedCentral
11.
go back to reference van Woerden HC, Ratier-Cruz A, Aleshinloye OB, Martinez-Giron R, Gregory C, Matthews IP: Association between protozoa in sputum and asthma: a case–control study. Respir Med. 2011, 105 (6): 877-884. 10.1016/j.rmed.2010.11.016.CrossRefPubMed van Woerden HC, Ratier-Cruz A, Aleshinloye OB, Martinez-Giron R, Gregory C, Matthews IP: Association between protozoa in sputum and asthma: a case–control study. Respir Med. 2011, 105 (6): 877-884. 10.1016/j.rmed.2010.11.016.CrossRefPubMed
12.
go back to reference Pirhonen I, Nevalainen A, Husman T, Pekkanen J: Home dampness, moulds and their influence on respiratory infections and symptoms in adults in Finland. Eur Respir J. 1996, 9 (12): 2618-2622. 10.1183/09031936.96.09122618.CrossRefPubMed Pirhonen I, Nevalainen A, Husman T, Pekkanen J: Home dampness, moulds and their influence on respiratory infections and symptoms in adults in Finland. Eur Respir J. 1996, 9 (12): 2618-2622. 10.1183/09031936.96.09122618.CrossRefPubMed
13.
go back to reference Amann RI, Krumholz L, Stahl DA: Fluorescent-oligonucleotide probing of whole cells for determinative, phylogenetic, and environmental studies in microbiology. J Bacteriol. 1990, 172 (2): 762-770.PubMedPubMedCentral Amann RI, Krumholz L, Stahl DA: Fluorescent-oligonucleotide probing of whole cells for determinative, phylogenetic, and environmental studies in microbiology. J Bacteriol. 1990, 172 (2): 762-770.PubMedPubMedCentral
14.
go back to reference Sogin ML, Gunderson JH: Structural diversity of eukaryotic small subunit ribosomal RNAs. Evolutionary implications. Ann N Y Acad Sci. 1987, 503: 125-139. 10.1111/j.1749-6632.1987.tb40603.x.CrossRefPubMed Sogin ML, Gunderson JH: Structural diversity of eukaryotic small subunit ribosomal RNAs. Evolutionary implications. Ann N Y Acad Sci. 1987, 503: 125-139. 10.1111/j.1749-6632.1987.tb40603.x.CrossRefPubMed
15.
go back to reference Lozupone C, Hamady M, Knight R: UniFrac–an online tool for comparing microbial community diversity in a phylogenetic context. BMC Bioinformatics. 2006, 7: 371-10.1186/1471-2105-7-371.CrossRefPubMedPubMedCentral Lozupone C, Hamady M, Knight R: UniFrac–an online tool for comparing microbial community diversity in a phylogenetic context. BMC Bioinformatics. 2006, 7: 371-10.1186/1471-2105-7-371.CrossRefPubMedPubMedCentral
16.
go back to reference Martin AP: Phylogenetic approaches for describing and comparing the diversity of microbial communities. Appl Environ Microbiol. 2002, 68 (8): 3673-3682. 10.1128/AEM.68.8.3673-3682.2002.CrossRefPubMedPubMedCentral Martin AP: Phylogenetic approaches for describing and comparing the diversity of microbial communities. Appl Environ Microbiol. 2002, 68 (8): 3673-3682. 10.1128/AEM.68.8.3673-3682.2002.CrossRefPubMedPubMedCentral
17.
go back to reference Gaitanis G, Magiatis P, Hantschke M, Bassukas ID, Velegraki A: The Malassezia genus in skin and systemic diseases. Clin Microbiol Rev. 2012, 25 (1): 106-141. 10.1128/CMR.00021-11.CrossRefPubMedPubMedCentral Gaitanis G, Magiatis P, Hantschke M, Bassukas ID, Velegraki A: The Malassezia genus in skin and systemic diseases. Clin Microbiol Rev. 2012, 25 (1): 106-141. 10.1128/CMR.00021-11.CrossRefPubMedPubMedCentral
18.
go back to reference Vasutov M, Urban A: Phylogenetic studies in Psathyrella focusing on sections Pennatae and Spadiceae–new evidence for the paraphyly of the genus. Mycol Res. 2008, 112 (10): 1153-1164. 10.1016/j.mycres.2008.04.005.CrossRef Vasutov M, Urban A: Phylogenetic studies in Psathyrella focusing on sections Pennatae and Spadiceae–new evidence for the paraphyly of the genus. Mycol Res. 2008, 112 (10): 1153-1164. 10.1016/j.mycres.2008.04.005.CrossRef
19.
go back to reference Pitkaranta M, Meklin T, Hyvarinen A, Paulin L, Auvinen P, Nevalainen A, Rintala H: Analysis of fungal flora in indoor dust by ribosomal DNA sequence analysis, quantitative PCR, and culture. Appl Environ Microbiol. 2008, 74 (1): 233-244. 10.1128/AEM.00692-07.CrossRefPubMed Pitkaranta M, Meklin T, Hyvarinen A, Paulin L, Auvinen P, Nevalainen A, Rintala H: Analysis of fungal flora in indoor dust by ribosomal DNA sequence analysis, quantitative PCR, and culture. Appl Environ Microbiol. 2008, 74 (1): 233-244. 10.1128/AEM.00692-07.CrossRefPubMed
20.
go back to reference Burr ML, Matthews IP, Arthur RA, Watson HL, Gregory CJ, Dunstan FD, Palmer SR: Effects on patients with asthma of eradicating visible indoor mould: a randomised controlled trial. Thorax. 2007, 62 (9): 767-772. 10.1136/thx.2006.070847.CrossRefPubMedPubMedCentral Burr ML, Matthews IP, Arthur RA, Watson HL, Gregory CJ, Dunstan FD, Palmer SR: Effects on patients with asthma of eradicating visible indoor mould: a randomised controlled trial. Thorax. 2007, 62 (9): 767-772. 10.1136/thx.2006.070847.CrossRefPubMedPubMedCentral
21.
go back to reference Quast C, Pruesse E, Yilmaz P, Gerken J, Schweer T, Yarza P, Peplies J, Glockner FO: The SILVA ribosomal RNA gene database project: improved data processing and web-based tools. Nucleic Acids Res. 2013, 41 (D1): D590-596. 10.1093/nar/gks1219.CrossRefPubMed Quast C, Pruesse E, Yilmaz P, Gerken J, Schweer T, Yarza P, Peplies J, Glockner FO: The SILVA ribosomal RNA gene database project: improved data processing and web-based tools. Nucleic Acids Res. 2013, 41 (D1): D590-596. 10.1093/nar/gks1219.CrossRefPubMed
22.
go back to reference Jacquet A: The role of the house dust mite-induced innate immunity in development of allergic response. Int Arch Allergy Immunol. 2010, 155 (2): 95-105.CrossRefPubMed Jacquet A: The role of the house dust mite-induced innate immunity in development of allergic response. Int Arch Allergy Immunol. 2010, 155 (2): 95-105.CrossRefPubMed
23.
go back to reference Jacquet A: Interactions of airway epithelium with protease allergens in the allergic response. Clin Exp Allergy. 2011, 41 (3): 305-311. 10.1111/j.1365-2222.2010.03661.x.CrossRefPubMed Jacquet A: Interactions of airway epithelium with protease allergens in the allergic response. Clin Exp Allergy. 2011, 41 (3): 305-311. 10.1111/j.1365-2222.2010.03661.x.CrossRefPubMed
Metadata
Title
Differences in fungi present in induced sputum samples from asthma patients and non-atopic controls: a community based case control study
Authors
Hugo Cornelis van Woerden
Clive Gregory
Richard Brown
Julian Roberto Marchesi
Bastiaan Hoogendoorn
Ian Price Matthews
Publication date
01-12-2013
Publisher
BioMed Central
Published in
BMC Infectious Diseases / Issue 1/2013
Electronic ISSN: 1471-2334
DOI
https://doi.org/10.1186/1471-2334-13-69

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