title: Monitoring the rumen pectinolytic bacteria Treponema saccharophilum using real-time PCR.
abstract: Treponema saccharophilum is a pectinolytic bacterium isolated from the bovine rumen. The abundance of this bacterium has not been well determined, reflecting the lack of a reliable and accurate detection method. To develop a rapid method for monitoring T. saccharophilum, we performed pyrosequencing of genomic DNA isolated from rumen microbiota to explore the 16S rRNA gene sequences of T. saccharophilum candidates. Species-specific primers were designed based on fifteen sequences of partial 16S rRNA genes generated through pyrosequencing with 97% or higher similarity with T. saccharophilum DSM2985 along with sequence from type strain. The relative abundance of T. saccharophilum was quantified in both in vitro and in vivo rumen systems with varied pectin-containing forages using real-time PCR. There was a clear association of T. saccharophilum with alfalfa hay, which contains more pectin than Chinese wild rye hay or corn stover. The relative abundance of T. saccharophilum was as high as 0.58% in vivo, comparable with the population density of other common rumen bacteria. It is recognized that T. saccharophilum plays an important role in pectin digestion in the rumen.
authors.affiliation: Institute of Dairy Science, MoE Key Laboratory of Molecular Animal Nutrition, College of Animal Sciences, Zhejiang University, Hangzhou, China.
authors.forename: Jing
authors.initials: J
authors.lastname: Liu
authors.forename: Jia-Kun
authors.initials: JK
authors.lastname: Wang
authors.forename: Wen
authors.initials: W
authors.lastname: Zhu
authors.forename: Yi-Yi
authors.initials: YY
authors.lastname: Pu
authors.forename: Le-Luo
authors.initials: LL
authors.lastname: Guan
authors.forename: Jian-Xin
authors.initials: JX
authors.lastname: Liu
mesh_terms: D000821:Animal Feed; D000818:Animals; D002417:Cattle / Q000382:microbiology; D017931:DNA Primers / Q000235:genetics; D004269:DNA, Bacterial / Q000235:genetics / Q000302:isolation & purification; D005260:Female; D008129:Lolium / Q000737:chemistry; D010368:Pectins / Q000378:metabolism*; D012336:RNA, Ribosomal, 16S / Q000235:genetics; D060888:Real-Time Polymerase Chain Reaction; D012417:Rumen / Q000382:microbiology*; D012680:Sensitivity and Specificity; D017422:Sequence Analysis, DNA; D014208:Treponema / Q000235:genetics / Q000302:isolation & purification*; D003313:Zea mays / Q000737:chemistry
chemical_list: D017931:DNA Primers; D004269:DNA, Bacterial; D012336:RNA, Ribosomal, 16S; D010368:Pectins
keywords: 16S rRNA gene; Treponema saccharophilum; forage; pectin