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Applied and Environmental Microbiology, February 1999, p. 381-388, Vol. 65, No. 2
Microbial and Enzymatic Technology
Group1 and
Environmental Bioengineering
Group,2 Biotechnology Research Institute,
National Research Council of Canada, Montreal, Quebec, Canada H4P
2R2
Received 26 August 1998/Accepted 4 November 1998
We attempted to mimic in small upflow anaerobic sludge bed (UASB)
bioreactors the metabolic association found in nature between methanogens and methanotrophs. UASB bioreactors were inoculated with
pure cultures of methanotrophs, and the bioreactors were operated by using continuous low-level oxygenation in order to favor
growth and/or survival of methanotrophs. Unlike the reactors in other
similar studies, the hybrid anaerobic-aerobic bioreactors which we used were operated synchronously, not sequentially. Here, emphasis was placed on monitoring various methanotrophic populations by
using classical methods and also a PCR amplification assay based on the
mmoX gene fragment of the soluble methane monooxygenase (sMMO). The following results were obtained: (i) under the
conditions used, Methylosinus sporium appeared to survive
better than Methylosinus trichosporium; (ii) the PCR method
which we used could detect as few as about 2,000 sMMO
gene-containing methanotrophs per g (wet weight) of granular
sludge; (iii) inoculation of the bioreactors with pure cultures of
methanotrophs contributed greatly to increases in the sMMO-containing
population (although the sMMO-containing population decreased gradually
with time, at the end of an experiment it was always at least 2 logs
larger than the initial population before inoculation); (iv) in
general, there was a good correlation between populations with the sMMO
gene and populations that exhibited sMMO activity; and (v) inoculation
with sMMO-positive cultures helped increase significantly the
proportion of sMMO-positive methanotrophs in reactors,
even after several weeks of operation under various regimes. At some
point, anaerobic-aerobic bioreactors like those described here
might be used for biodegradation of various chlorinated pollutants.
0099-2240/99/$00.00+0
Monitoring Methanotrophic Bacteria in Hybrid Anaerobic-Aerobic
Reactors with PCR and a Catabolic Gene Probe
*
Corresponding author. Mailing address: Microbial and
Enzymatic Technology Group, Biotechnology Research Institute, National Research Council of Canada, 6100 Royalmount Avenue, Montreal, Quebec,
Canada H4P 2R2. Phone: (514) 496-6186. Fax: (514) 496-5485. E-mail:
Denis.Groleau{at}nrc.ca.
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