Molecular monitoring and isolation of previously uncultured bacterial strains from the sheep rumen.

Applied and Environmental Microbiology
S KoikeY Kobayashi

Abstract

To estimate the contribution of uncultured bacterial groups to fiber degradation, we attempted to retrieve both ecological and functional information on uncultured groups in the rumen. Among previously reported uncultured bacteria, fiber-associated groups U2 and U3, belonging to the low-GC Gram-positive bacterial group, were targeted. PCR primers and fluorescence in situ hybridization (FISH) probe targeting 16S rRNA genes or rRNA were designed and used to monitor the distribution of targets. The population size of group U2 in the rumen was as high as 1.87%, while that of group U3 was only 0.03%. Strong fluorescence signals were observed from group U2 cells attached to plant fibers in the rumen. These findings indicate the ecological significance of group U2 in the rumen. We succeeded in enriching group U2 using rumen-incubated rice straw as the inoculum followed by incubation in an appropriate medium with an agent inhibitory for Gram-negative bacteria. Consequently, we successfully isolated two strains, designated B76 and R-25, belonging to group U2. Both strains were Gram-positive short rods or cocci that were 0.5 to 0.8 mum in size. Strain B76 possessed xylanase and alpha-l-arabinofuranosidase activity. In particular, the xyl...Continue Reading

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Citations

Oct 4, 2012·Animal : an International Journal of Animal Bioscience·D P MorgaviG T Attwood
Jun 14, 2013·Animal : an International Journal of Animal Bioscience·S C LeahyG T Attwood
Mar 13, 2014·Animal Science Journal = Nihon Chikusan Gakkaihō·Satoshi KoikeYasuo Kobayashi
Nov 23, 2013·Animal Science Journal = Nihon Chikusan Gakkaihō·Takumi ShinkaiYasuo Kobayashi
Jul 6, 2014·Microbial Biotechnology·Christopher J CreeveySinead C Leahy
Dec 3, 2013·FEMS Microbiology Ecology·Jing LiuJian-Xin Liu
Feb 28, 2017·The Journal of General and Applied Microbiology·Jinqiang HuangYuzhu Luo
Apr 25, 2012·Folia Microbiologica·Jaime A RoseroJan Kopečný

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