Project description:Sequencing of 16S ribosomal RNA (rRNA) gene, which has improved the characterization of microbial community, has made it possible to detect a low level Helicobacter pylori (HP) sequences even in HP-negative subjects which were determined by a combination of conventional methods. This study was conducted to obtain a cutoff value for HP colonization in gastric mucosa biopsies and gastric juices by the pyrosequencing method. Corresponding author: Department of Internal Medicine, Seoul National University Bundang Hospital, Seoungnam, Gyeonggi-do, Korea; Department of Internal Medicine and Liver Research Institute, Seoul National University College of Medicine, Seoul, Korea (Tel., +82-31-787-7008; e-mail, nayoungkim49@empas.com). Microbial DNA from gastric mucosal samples [gastric antrum (n=63, mucosal biopsy), follow-up sample on gastric antrum (n=16, mucosal biopsy), and gastric body (n=18, mucosal biopsy)] and gastric juices (n=4, not mucosal biopsy) was amplified by nested PCR using universal bacterial primers, and the 16S rRNA genes were pyrosequenced.
Project description:Sequencing of 16S ribosomal RNA (rRNA) gene, which has improved the characterization of microbial community, has made it possible to detect a low level Helicobacter pylori (HP) sequences even in HP-negative subjects which were determined by a combination of conventional methods. This study was conducted to obtain a cutoff value for HP colonization in gastric mucosa biopsies and gastric juices by the pyrosequencing method. Corresponding author: Nayoung Kim, Department of Internal Medicine, Seoul National University Bundang Hospital, Seoungnam, Gyeonggi-do, Korea; Department of Internal Medicine and Liver Research Institute, Seoul National University College of Medicine, Seoul, Korea (Tel., +82-31-787-7008; e-mail, nayoungkim49@empas.com).
Project description:Two potato cultivars, Russet Burbank and Bionta, were inoculated with three different endophytes containing different AHL types. The impact of the endophytes to the different cultivars was measured by gene expression analysis with a customized microarray B. phytofirmans type strain PsJN was originally isolated as a contaminant from surface-sterilized, Glomus vesculiferum-infected onion roots (Nowak et al., 1998), whereas strain P6 RG6-12 was isolated from the rhizosphere of a grassland in the Netherlands (Salles et al., 2006). This strain was selected based on its similarity to strain PsJN based on 16S rRNA gene homology, and similar phenotypic features. Both strains were generally cultivated on King's medium (King et al., 1954). For the mutant AHL to the strain B. phytofirmans PsJN a quorum quenching approach as described by Wopperer et al., 2006 was employed. Plasmid pMLBAD-aiiA, which contains aiiA, the Bacillus sp. 240B1 lactonase gene, was transferred to B. phytofirmans PsJN by triparental mating as described by de Lorenzo and Timmis (1994). 2 cultivars, 3 endophytes
Project description:The effects of increasing addition of green tea in dietary changes the bacterial populations in broiler ileum were evaluated. Four hundreds of AA broilers were randomly assigned to four groups with green tea addition of 0, 0.5, 1 and 2 percent in the diet. The body weight showed no difference but a digital increase positively correlated with addition of green tea. The content of green tea had a linear effect of lengthening the ileum villi. The barcoded DNA pyrosequencing method was used to reveal 15 phyla, 1157phylotypes and 3098 16S operational taxonomic units (OTUs). The most predominant bacterial phyla were Firmicutes (56.89%), Actinobacteria (30.58%), Proteobacteria (8.61%) and Bacteroidetes (2.72%). As the proportion of additional green tea increased, the abundance of phylum Actinobacteria (p=0.003) and Proteobacteria (p=0.049) almost linearly increased, while the proportion of Firmicutes (p=0.027) linearly decreased. Only 2 OTUs were significantly affected by the increased additive, Corynebacteriaceae (p=0.011) and Staphylococcaceae (p= 0.006). Triplot analysis suggested that the dominant phyla of Verrucomicrobia, TM7 and Actinobacteria were clearly related to the addition of green tea. Moreover, green tea addition influenced the construction of microbiota, and lengthened the villus in ileum by Monte Carlo permutation test. These findings provide a new understanding of the ileal microbial ecology, which may be useful in modulating the gut microbiome, and also the proper usage of powdered green tea.