PML14T, was isolated from the sludge of a thermophilic
anaerobic digester treating municipal solid waste and
sewage and its characteristics were determined.
Tepidimicrobium ferriphilum DSM 16624T was purchased
from the DSMZ. Strain PML14T was isolated in prereduced
peptone-yeast extract-glucose (PYG) medium
(Holdeman et al., 1977) by serial dilution and the
Hungate roll-tube technique (Hungate, 1969). Single
colonies were picked, transferred to the same broth and
incubated at 60 uC for 2 days. The roll-tube procedure was
repeated several times until a pure culture was obtained.
The purity of strain PML14T was confirmed by observing
similar morphologies of colonies on agar and of cells under
a microscope. Routine cultivation was performed with
PYG broth in anaerobic tubes sealed with butyl rubber
stoppers under a gaseous atmosphere of N2
(1.016105 Pa).
Spore staining was performed using the Schaeffer and
Fulton staining method as described by Barrow & Feltham
(1993). Substrate utilization tests, light microscopy and