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Microbiological Community Structure of the Biofilm of a Methanol-Fed, Marine Denitrification System, and Identification of the Methanol-Utilizing Microorganisms

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Abstract

We demonstrated in a previous study that the biofilm of the methanol-fed fluidized marine denitrification reactor at the Montreal Biodome was composed of at least 15 bacterial phylotypes. Among those were 16S ribosomal RNA (rDNA) gene sequences affiliated to Hyphomicrobium spp., and Methylophaga spp.; the latter made up 70% of a clone library. By using fluorescent in situ hybridization (FISH), we investigated the structure of the biofilm during the colonization process in the denitrification reactor by targeting most of the bacterial families that the 16S rDNA gene library suggested would occur in the biofilm. Our results revealed that gamma-Proteobacteria (mostly Methylophaga spp.) accounted for up to 79% of the bacterial population, confirming the abundance of Methylophaga spp. within the biofilm. alpha-Proteobacteria represented 27–57% of the population, which included Hyphomicrobium spp. that appeared after 20 days of colonization and represented 7–8% of the population. We noticed a great abundance and diversity of eukaryotic cells, which made up 20% of the biomass at the beginning of the colonization but decreased to 3–5% in the mature biofilm. We then used FISH combined with microautoradiography (MAR–FISH) to identify the methylotrophs in the biofilm. The results showed that alpha-Proteobacteria used 14C methanol in the presence of nitrate, suggesting their involvement in denitrification. Despite their abundance, Methylophaga spp. did not assimilate methanol under those conditions.

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Acknowledgments

This research was supported by the Montreal Biodome, by a scholarship to N. Labbé from the Fondation Armand-Frappier and by two grants to R. Villemur from the Natural Sciences and Engineering Research Council of Canada. We are grateful to Rachel Léger, Head of Live Collections and Research, for her continuous support. We also wish to thank Jean Bouvrette, Mec. Eng. and Maintenance and Operation Manager, and the engineering staff of the Biodome for their technical help and cooperation. Brian Colwill, C.Tr., kindly revised the English style and grammar.

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Authors and Affiliations

  1. INRS – Institut Armand-Frappier, 531 boul. des Prairies, Laval, Canada, H7V 1B7

    Normand Labbé, Véronique Laurin, Pierre Juteau & Richard Villemur

  2. Biodôme de Montréal, 4777 Pierre-De Coubertin, Montréal, Canada, H1V 1B3

    Serge Parent

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  1. Normand Labbé
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  2. Véronique Laurin
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Correspondence to Richard Villemur.

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Labbé, N., Laurin, V., Juteau, P. et al. Microbiological Community Structure of the Biofilm of a Methanol-Fed, Marine Denitrification System, and Identification of the Methanol-Utilizing Microorganisms. Microb Ecol 53, 621–630 (2007). https://doi.org/10.1007/s00248-006-9168-z

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  • DOI: https://doi.org/10.1007/s00248-006-9168-z

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