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Bacterial Diversity in a Marine Methanol-Fed Denitrification Reactor at the Montreal Biodome, Canada

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Abstract

The bacterial biota of a methanol-fed denitrification reactor used to treat seawater at the Montreal Biodome were investigated using culture-dependent and molecular biology methods. The microbiota extracted from the reactor carriers were cultivated on three media. Three isolate types were recovered and their 16S ribosomal DNA (rDNA) genes were determined. The analysis showed that the isolate types were related to α-Proteobacteria. They are members of the Hyphomicrobium and Paracoccus genera and the Phyllobacteriaceae family. Uncultured bacteria were identified through a 16S rDNA library generated from total DNA extracted from the microbiota. Clones were screened for different restriction profiles and for different DGGE (denaturing gradient gel electrophoresis) migration profiles. More than 70% of clones have the same restriction profile, and the sequence of representative clones showed a relation with the Methylophaga members of the Piscirickettsia family (γ-Proteobacteria). Sequences from other profiles were related to bacterial species involved in denitrification. The number of species in the denitrification reactor was estimated at 15. Bacterial colonization on newly added carriers in the denitrification reactor was monitored by PCR-DGGE. The DGGE migration profiles evolved during the first 5 weeks and then remained essentially unchanged. PCR-DGGE was also used to monitor the microbial profiles in various aquarium locations. As expected, bacterial populations differed from one location to another, except for the sand and trickling filters which presented similar DGGE migration profiles.

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Acknowledgements

This research was supported by a grant to R. Villemur from the Natural Sciences and Engineering Research Council of Canada, by funding from the Montreal Biodome, and by a scholarship to N. Labbé from the INRS-Institut Armand-Frappier. 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 Operations 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. EcoLink, Bioflow, and NuPac are registered trademarks.

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Authors and Affiliations

  1. INRS—Institut Armand-Frappier, 531 Boul. des Prairies, Laval H7V 1B7, Canada

    N. Labbé, P. Juteau & R. Villemur

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

    S. Parent

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  1. N. Labbé
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Correspondence to R. Villemur.

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Labbé, N., Juteau, P., Parent, S. et al. Bacterial Diversity in a Marine Methanol-Fed Denitrification Reactor at the Montreal Biodome, Canada . Microb Ecol 46, 12–21 (2003). https://doi.org/10.1007/s00248-002-1056-6

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