PMID- 17209077
OWN - NLM
STAT- MEDLINE
DCOM- 20070410
LR  - 20191210
IS  - 0099-2240 (Print)
IS  - 1098-5336 (Electronic)
IS  - 0099-2240 (Linking)
VI  - 73
IP  - 5
DP  - 2007 Mar
TI  - Localization of ruminal cellulolytic bacteria on plant fibrous materials as 
      determined by fluorescence in situ hybridization and real-time PCR.
PG  - 1646-52
AB  - To visualize and localize specific bacteria associated with plant materials, a 
      new fluorescence in situ hybridization (FISH) protocol was established. By using 
      this protocol, we successfully minimized the autofluorescence of orchard grass 
      hay and detected rumen bacteria attached to the hay under a fluorescence 
      microscope. Real-time PCR assays were also employed to quantitatively monitor the 
      representative fibrolytic species Fibrobacter succinogenes and Ruminococcus 
      flavefaciens and also total bacteria attached to the hay. F. succinogenes was 
      found firmly attached to not only the cut edges but also undamaged inner surfaces 
      of the hay. Cells of phylogenetic group 1 of F. succinogenes were detected on 
      many stem and leaf sheath fragments of the hay, even on fragments on which few 
      other bacteria were seen. Cells of phylogenetic group 2 of F. succinogenes were 
      often detected on hay fragments coexisting with many other bacteria. On the basis 
      of 16S rRNA gene copy number analysis, the numbers of bacteria attached to the 
      leaf sheaths were higher than those attached to the stems (P<0.05). In addition, 
      R. flavefaciens had a greater tendency than F. succinogenes to be found on the 
      leaf sheath (P<0.01) with formation of many pits. F. succinogenes, particularly 
      phylogenetic group 1, is suggested to possibly play an important role in fiber 
      digestion, because it is clearly detectable by FISH and is the bacterium with the 
      largest population size in the less easily degradable hay stem.
FAU - Shinkai, Takumi
AU  - Shinkai T
AD  - Graduate School of Agriculture, Hokkaido University, Kita-ku, Sapporo-shi 
      060-8589, Japan.
FAU - Kobayashi, Yasuo
AU  - Kobayashi Y
LA  - eng
PT  - Evaluation Study
PT  - Journal Article
DEP - 20070105
PL  - United States
TA  - Appl Environ Microbiol
JT  - Applied and environmental microbiology
JID - 7605801
RN  - 0 (DNA, Bacterial)
RN  - 0 (RNA, Ribosomal, 16S)
RN  - 9004-34-6 (Cellulose)
SB  - IM
MH  - Animals
MH  - Cellulose/*metabolism
MH  - DNA, Bacterial/analysis
MH  - Dactylis/*microbiology
MH  - Fibrobacter/classification/genetics/isolation & purification
MH  - In Situ Hybridization, Fluorescence/*methods
MH  - Male
MH  - Plant Leaves/microbiology
MH  - Plant Stems/microbiology
MH  - Polymerase Chain Reaction/*methods
MH  - RNA, Ribosomal, 16S/genetics
MH  - Rumen/*microbiology
MH  - Ruminococcus/classification/genetics/isolation & purification
MH  - Sheep/microbiology
PMC - PMC1828787
EDAT- 2007/01/09 09:00
MHDA- 2007/04/11 09:00
PMCR- 2007/07/01
CRDT- 2007/01/09 09:00
PHST- 2007/01/09 09:00 [pubmed]
PHST- 2007/04/11 09:00 [medline]
PHST- 2007/01/09 09:00 [entrez]
PHST- 2007/07/01 00:00 [pmc-release]
AID - AEM.01896-06 [pii]
AID - 1896-06 [pii]
AID - 10.1128/AEM.01896-06 [doi]
PST - ppublish
SO  - Appl Environ Microbiol. 2007 Mar;73(5):1646-52. doi: 10.1128/AEM.01896-06. Epub 
      2007 Jan 5.