Differentiation of Methanosaeta concilii and Methanocarcina barkeri in anaerobic mesophilic granular sludge by fluorescent in situ hybridization and confocal scanning laser microscopy

Journal Article · Sat May 01 04:00:00 EDT 1999 · Applied and Environmental Microbiology

OSTI ID:346837

Rocheleau, S; Greer, C W; Cantin, C; Laramee, L; Guiot, S R ^[1]; Lawrence, J R ^[2]

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1. National Research Council Canada, Montreal, Quebec (Canada). Biotechnology Research Inst.
2. National Water Research Inst., Saskatoon, Saskatchewan (Canada)

Oligonucleotide probes, designed from genes coding for 16S rRNA, were developed to differentiate Methanosaeta concilii, Methanosarcina barkeri, and mesophilic methanogens. All M. concilii oligonucleotide probes (designated MS1, MS2, and MS5) hybridized specifically with the target DNA, but MS5 was the most specific M. concilii oligonucleotide probe. Methanosarcina barkeri oligonucleotide probes (designated MB1, MB3, and MB4) hybridized with different Methanosarcina species. The MB4 probe specifically detected Methanosarcina barkeri, and the MB3 probe detected the presence of al mesophilic Methanosarcina species. These new oligonucleotide probes facilitated the identification, localization, and quantification of the specific relative abundance of M. concilii and Methanosarcina barkeri, which play important roles in methanogenesis. The combined use of fluorescent in situ hybridization with confocal scanning laser microscopy demonstrated that anaerobic granule topography depends on granule origin and feeding. Protein-fed granules showed no layered structure with a random distribution of M. concilii. In contrast, a layered structure developed in methanol-enriched granules, where M. barkeri growth was induced in an outer layer. This outer layer was followed by a layer composed of M. concilii, with an inner core of M. concilii and other bacteria.

• OSTI ID: 346837
• Journal Information: Applied and Environmental Microbiology, Journal Name: Applied and Environmental Microbiology Journal Issue: 5 Vol. 65; ISSN AEMIDF; ISSN 0099-2240
• Country of Publication: United States
• Language: English

Related Subjects

09 BIOMASS FUELS
ANAEROBIC CONDITIONS
BIOSYNTHESIS
DNA HYBRIDIZATION
GENETIC VARIABILITY
METHANE
METHANOGENIC BACTERIA
OLIGONUCLEOTIDES