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Methanobrevibacter millerae sp. nov. and Methanobrevibacter olleyae sp. nov., methanogens from the ovine and bovine rumen that can utilize formate for growth

¹ CSIRO Minerals, cnr Brand and Townsing Drives, Bentley, Western Australia 6102, Australia
² Australian Food Safety Centre, University of Tasmania, Private Bag 54, Hobart, Tasmania 7001, Australia
³ CPSU, Level 1, 40 Brisbane Avenue, Barton, Australian Capital Territory 2600, Australia
⁴ CSIRO Livestock Industries, Underwood Avenue, Floreat Park, Western Australia 6014, Australia
⁵ CSIRO Livestock Industries, 306 Carmody Rd, St Lucia, Queensland 4067, Australia

Correspondence
Suzanne Rea
Suzy.Rea{at}csiro.au

Four formate-utilizing methanogens were isolated from ovine (strain KM1H5-1P^T) and bovine (strains AK-87, OCP and ZA-10^T) rumen contents. Based on 16S rRNA gene sequence analysis, the methanogen strains were found to belong to the order Methanobacteriales in the genus Methanobrevibacter. Strains ZA-10^T and KM1H5-1P^T gained energy for growth by the reduction of CO₂ to CH₄ using H₂ or formate exclusively as electron donors. Increasing formate concentrations to 220 mM in batch cultures increased the growth of strain KM1H5-1P^T but did not affect the growth of strain ZA-10^T. Substrate specificity and resistance to cell-wall lysis supported the affiliation of the strains to the genus Methanobrevibacter. Strains ZA-10^T and KM1H5-1P^T showed 16S rRNA gene sequence similarity of 98.0 and 98.6 % to their closest recognized relatives, Methanobrevibacter thaueri CW^T and Methanobrevibacter ruminantium M1^T, respectively. DNA–DNA hybridization experiments indicated that the strains were not affiliated at the species level to their closest recognized relatives, with DNA reassociation values of only 28 % between strains ZA-10^T and Methanobrevibacter thaueri CW^T and <25 % between strains KM1H5-1P^T and Methanobrevibacter ruminantium M1^T. Based on the data presented, the new strains are considered to represent two novel species of the genus Methanobrevibacter, for which the names Methanobrevibacter millerae sp. nov. (type strain ZA-10^T=DSM 16643^T=OCM 820^T) and Methanobrevibacter olleyae sp. nov. (type strain KM1H5-1P^T=DSM 16632^T=OCM 841^T) are proposed.

Details of growth parameters and DNA–DNA hybridization and 16S rRNA gene sequence similarity values for strains ZA-10^T and M1H5-1P^T in comparison with Methanobrevibacter reference strains are available as supplementary material in IJSEM Online.

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