Project description:An Easy Operating Pathogen Microarray (EOPM) was designed to detect almost all known pathogens and related species based on their genomic sequences. For effective identification of pathogens from EOPM data, a statistical enrichment algorithm has been proposed and further implemented in a user-friendly interface. A microarray was designed with probes for vertebrate-infecting virus sequences in EMBL, 18S rRNA fungi and parasite sequences from EMBL, and 16S rRNA sequences of bacteria from RDP, synthesized on the Agilent platform. The array was tested using 2 color dyes on cultured microbes and on clinical samples from sick and healthy people, looking for differences in clinically ill people compared to a number of healthy "controls".
Project description:Iron-rich pelagic aggregates (iron snow) were collected directly onto silicate glass filters using an electronic water pump installed below the redoxcline. RNA was extracted and library preparation was done using the NEBNext Ultra II directional RNA library prep kit for Illumina. Data was demultiplied by GATC sequencing company and adaptor was trimmed by Trimgalore. After trimming, data was processed quality control by sickle and mRNA/rRNA sequences were sorted by SortmeRNA. mRNA sequences were blast against NCBI-non redundant protein database and the outputs were meganized in MEGAN to do functional analysis. rRNA sequences were further sorted against bacterial/archeal 16S rRNA, eukaryotic 18S rRNA and 10,000 rRNA sequences of bacterial 16S rRNA, eukaryotic 18S rRNA were subset to do taxonomy analysis.
Project description:The community composition (in terms of abundance, distribution and contribution of diverse clades) of bacteria involved in nitrogen transformations in the oxygen minimum zones may be related to the rates of fixed N loss in these systems. The abundance of both denirifying and anammox bacteria, and the assemblage composition of denitrifying bacteria were investigated in the Eastern Tropical South Pacific and the Arabian Sea using assays based on molecular markers for the two groups of bacteria. The abundance and distribution of bacteria associated with the fixed N removal processes denitrification and anammox were investigated using quantitative PCR for genes encoding nitrite reductase (nirK and nirS) in denitrifying bacteria and hydrazine oxidase(hzo) and 16S rRNA genesin anammox bacteria. All of these genes had depth distributions with maxima associated with the secondary nitrite maximum in low oxygen waters. NirS was mch more abundant than nirK, and much more abundant than the 16S rRNA gene from anammox bacteria. The ratio of hzo:16S rRNA for anammox was low and variable implying greater unexplored diversity in the the hzo gene. Assemblage composition of the abundant nirS-type denitrifiers was evaluated using a funcitonal gene microarray. Of the nirS archetypes represented on the microarray, very few occurred speficically in one region or depth interval, but the assemblages varied significantly. Community composition of denitrifiers based on microarray analysis of the nirS gene was most different between geographical regions. Within each region, the surface layer and OMZ assemblages clustered distinctly. Thus, in addition to spatial and temporal variation in denitrificaiton and anammox rates, both microbial abundance and community composition also vary between OMZ regions and depths. Two color array (Cy3 and Cy5): the universal standard 20-mer oligo is printed to the slide with a 70-mer oligo (an archetype). Environmental DNA sequences (fluoresced with Cy3) within 15% of the 70-mer conjugated to a 20-mer oligo (fluoresced with Cy5) complementary to the universal standard will bind to the oligo probes on the array. Signal is the ratio of Cy3 to Cy5. Three replicate probes were printed for each archetype. Two replicate arrays were run on duplicate targets.
Project description:Here we report 16s rRNA data in gut microbiota of hepatocellular carcinoma (HCC) patients with HBV induced HCC (HBVC) and non-HBV induced HCC (NHBVC) compared with healthy volunteers. A total of 2047 operational taxonomic units (OTUs) were identified in the sequence data. Our data shows that the NHBVC patients harbor lower anti-inflammatory bacteria and more pro-inflammatory bacteria, while the HBVC patients harbor more anti-inflammatory bacteria.
Project description:We characterized the bacterial diversity of chlorinated drinking water from three surface water treatment plants supplying the city of Paris, France. For this purpose, we used serial analysis of V6 ribosomal sequence tag (SARST-V6) to produce concatemers of PCR-amplified ribosomal sequence tags (RSTs) from the V6 hypervariable region of the 16S rRNA gene for sequence analysis. Using SARST-V6, we obtained bacterial profiles for each drinking water sample, demonstrating a strikingly high degree of biodiversity dominated by a large collection of low-abundance phylotypes. In all water samples, between 57.2-77.4% of the sequences obtained indicated bacteria belonging to the Proteobacteria phylum. Full-length 16S rDNA sequences were also generated for each sample, and comparison of the RSTs with these sequences confirmed the accurate assignment for several abundant bacterial phyla identified by SARST-V6 analysis, including members of unclassified bacteria, which account for 6.3-36.5% of all V6 sequences. These results suggest that these bacteria may correspond to a common group adapted to drinking water systems. The V6 primers used were subsequently evaluated with a computer algorithm to assess their hybridization efficiency. Potential errors associated with primer-template mismatches and their impacts on taxonomic group detection were investigated. The biodiversity present in all three drinking water samples suggests that the bacterial load of the drinking water leaving treatment plants may play an important role in determining the downstream community dynamics of water distribution networks. 3 different drinking water samples (Orly, Ivry, Joinville drinking water sample)
Project description:Given the criticle role of gut bacteria involve in number of diseases, the gut microbiota from young and aged people were estimated using 16s rRNA next-generation sequencing. This study will benefit to identify the role of gut bacteria on the pathegenic mechasim of aging relative diseases.
Project description:A combination of shotgun metaproteomics and 16S rRNA gene pyrosequencing wasused to identify potential functional pathways and key microorganisms involved in long-chain fatty acids (LCFA) anaerobic conversion. Microbial communities degrading saturated- and unsaturated-LCFA were compared. Archaeal communities were mainly composed of Methanosaeta, Methanobacterium and Methanospirillum species, both in stearate (saturated C18:0) and oleate (mono-unsaturated C18:1) incubations. Over 80% of the 16S rRNA gene sequences clustered within the Methanosaeta genus, which is in agreement with the high number of proteins assigned to this group (94%). Archaeal proteins related with methane metabolism were highly expressed. Bacterial communities were rather diverse and the composition dissimilar between incubations with saturated- and unsaturated-LCFA. Stearate-degrading communities were enriched in Deltaproteobacteria (34% of the assigned sequences), while microorganisms clustering within the Synergistia class were more predominant in oleate incubation (25% of the assigned sequences). Bacterial communities were diverse and active, given by the high percentage of proteins related with mechanisms of energy production. Several proteins were assigned to syntrophic bacteria, emphasizing the importance of the interactions between acetogens and methanogens in energy exchange and formation in anaerobic LCFA-rich environments.
Project description:A combination of shotgun metaproteomics and 16S rRNA gene pyrosequencing wasused to identify potential functional pathways and key microorganisms involved in long-chain fatty acids (LCFA) anaerobic conversion. Microbial communities degrading saturated- and unsaturated-LCFA were compared. Archaeal communities were mainly composed of Methanosaeta, Methanobacterium and Methanospirillum species, both in stearate (saturated C18:0) and oleate (mono-unsaturated C18:1) incubations. Over 80% of the 16S rRNA gene sequences clustered within the Methanosaeta genus, which is in agreement with the high number of proteins assigned to this group (94%). Archaeal proteins related with methane metabolism were highly expressed. Bacterial communities were rather diverse and the composition dissimilar between incubations with saturated- and unsaturated-LCFA. Stearate-degrading communities were enriched in Deltaproteobacteria (34% of the assigned sequences), while microorganisms clustering within the Synergistia class were more predominant in oleate incubation (25% of the assigned sequences). Bacterial communities were diverse and active, given by the high percentage of proteins related with mechanisms of energy production. Several proteins were assigned to syntrophic bacteria, emphasizing the importance of the interactions between acetogens and methanogens in energy exchange and formation in anaerobic LCFA-rich environments.