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Applied and Environmental Microbiology, August 2000, p. 3608-3615, Vol. 66, No. 8
Department of Environmental Systems
Engineering, Nagaoka University of Technology, Nagaoka, Niigata
940-2188,1 and National Institute of
Bioscience and Human-Technology, Agency of Industrial Science and
Technology, Tsukuba, Ibaraki 305-8566,2 Japan
Received 3 February 2000/Accepted 29 May 2000
The thermophilic, anaerobic, propionate-oxidizing bacterial
populations present in the methanogenic granular sludge in a
thermophilic (55°C) upflow anaerobic sludge blanket reactor were
studied by cultivation and in situ hybridization analysis. For
isolation of propionate-degrading microbes, primary enrichment was made with propionate as the sole energy source at 55°C. After several attempts to purify the microbes, a thermophilic, syntrophic,
propionate-oxidizing bacterium, designated strain SI, was isolated in
both pure culture and coculture with Methanobacterium
thermoautotrophicum. Under thermophilic (55°C) conditions,
strain SI oxidized propionate, ethanol, and lactate in coculture with
M. thermoautotrophicum. In pure culture, the isolate was
found to ferment pyruvate. 16S ribosomal DNA sequence analysis revealed
that the strain was relatively close to members of the genus
Desulfotomaculum, but it was only distantly related to any
known species. To elucidate the abundance and spatial distribution of
organisms of the strain SI type within the sludge granules, a 16S
rRNA-targeted oligonucleotide probe specific for strain SI was
developed and applied to thin sections of the granules. Fluorescence in
situ hybridization combined with confocal laser scanning microscopy
revealed that a number of rod-shaped cells were present in the middle
and inner layers of the thermophilic granule sections and that they
formed close associations with hydrogenotrophic methanogens. They
accounted for approximately 1.1% of the total cells in the sludge.
These results demonstrated that strain SI was one of the significant
populations in the granular sludge and that it was responsible for
propionate oxidation in the methanogenic granular sludge in the reactor.
0099-2240/00/$04.00+0
Copyright © 2000, American Society for Microbiology. All rights reserved.
Cultivation and In Situ Detection of a Thermophilic
Bacterium Capable of Oxidizing Propionate in Syntrophic Association
with Hydrogenotrophic Methanogens in a Thermophilic Methanogenic
Granular Sludge
*
Corresponding author. Mailing address: Department of
Environmental Systems Engineering, Nagaoka University of Technology, Kamitomioka 1603-1, Nagaoka, Niigata 940-2188, Japan. Phone:
81-258-47-9623. Fax: 81-258-47-9623. E-mail:
skgc{at}vos.nagaokaut.ac.jp.
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