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Applied and Environmental Microbiology, September 2009, p. 5445-5450, Vol. 75, No. 17
0099-2240/09/$08.00+0     doi:10.1128/AEM.00726-09
Copyright © 2009, American Society for Microbiology. All Rights Reserved.

Culture-Independent Characterization of Archaeal Biodiversity in Swine Confinement Building Bioaerosols{triangledown}

Benjamin Nehmé,1,2,{dagger} Yan Gilbert,1,2,{dagger} Valérie Létourneau,1,2 Robert J. Forster,3 Marc Veillette,1 Richard Villemur,4 and Caroline Duchaine1*

Centre de Recherche, Institut Universitaire de Cardiologie et de Pneumologie de Québec, Québec, Quebec, Canada,1 Département de Biochimie et de Microbiologie, Faculté des Sciences et de Génie, Université Laval, Québec, Quebec, Canada,2 Agriculture and Agri-Food Canada, Lethbridge, Alberta, Canada,3 INRS-Institut Armand-Frappier, Laval, Quebec, Canada4

Received 30 March 2009/ Accepted 20 June 2009

It was previously demonstrated that microbial communities of pig manure were composed of both bacteria and archaea. Recent studies have shown that bacteria are aerosolized from pig manure, but none have ever focused on the airborne archaeal burden. We sought here to develop and apply molecular ecology approaches to thoroughly characterize airborne archaea from swine confinement buildings (SCBs). Eight swine operations were visited, twice in winter and once during summer. Institute of Occupational Medicine cassettes loaded with 25-mm gelatin filters were used to capture the inhalable microbial biomass. The total genomic DNA was extracted and used as a template for PCR amplification of the archaeal 16S rRNA gene. High concentrations of archaea were found in SCB bioaerosols, being as high as 108 16S rRNA gene copies per cubic meter of air. Construction and sequencing of 16S rRNA gene libraries revealed that all sequences were closely related to methanogenic archaea, such as Methanosphaera stadtmanae (94.7% of the archaeal biodiversity). Archaeal community profiles were compared by 16S rRNA gene denaturing gradient gel electrophoresis. This analysis showed similar fingerprints in each SCB and confirmed the predominance of methanogenic archaea in the bioaerosols. This study sheds new light on the nature of bioaerosols in SCBs and suggests that archaea are also aerosolized from pig manure.

* Corresponding author. Mailing address: Centre de Recherche, Hôpital Laval, 2725, Chemin Ste.-Foy, Québec, Quebec G1V 4G5, Canada. Phone: (418) 656-8711, ext. 5837. Fax: (418) 656-4509. E-mail: caroline.duchaine{at}bcm.ulaval.ca

{triangledown} Published ahead of print on 26 June 2009.

{dagger} B.N. and Y.G. contributed equally to this study.

Applied and Environmental Microbiology, September 2009, p. 5445-5450, Vol. 75, No. 17
0099-2240/09/$08.00+0     doi:10.1128/AEM.00726-09
Copyright © 2009, American Society for Microbiology. All Rights Reserved.





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