PMC:4158115 / 8070-10021 JSONTXT

{"target":"https://pubannotation.org/docs/sourcedb/PMC/sourceid/4158115","sourcedb":"PMC","sourceid":"4158115","source_url":"http://www.ncbi.nlm.nih.gov/pmc/4158115","text":"2.4. Examination of C. concisus Growth under Anaerobic and Microaerobic Conditions in the Presence of Different Concentrations of H2 \nP3UCO-S1 strain was used for this experiment. P3UCO-S1 strain is an oral strain previously isolated from a patient with UC [6, 16]. In a previous study of analysis of six housekeeping genes, we showed that the six housekeeping genes of P3UCO-S1 strain were identical to the strain isolated from intestinal biopsies of the same patient (P3UCB-S1) suggesting that this oral C. concisus strain was able to colonize the intestinal tract. Given this, we have decided to use P3UCO-S1 strain in this part of the study.\nP3UCO-S1 strain was first cultured on a HBA plate under Oricon condition for 48 hours at 37°C. Following this, bacterial cells were collected and suspended into PBS and the OD600 was adjusted to 0.05. The bacterial suspension (30 μL) was inoculated onto 18 HBA plates. The plates were incubated under either anaerobic conditions or microaerobic conditions containing various conditions of H2 (three plates in each condition). Anaerobic and microaerobic conditions were generated using gas-generation system AN25A and gas-generation system CN25A, respectively (Oxoid). Hydrogen gas was supplemented by including 0.021 g, 0.042 g, or 0.083 g of sodium borohydride and 10 mL of H2O in a container placed in a 2.5 L jar, respectively, which generated 2.5%, 5.0%, and 10.0% of H2, respectively. H2 gas was generated by the chemical reaction NaBH4 + 4H2O = 4H2 + NaB(OH)4.\nAfter a period of 48 hours of incubation at 37°C, C. concisus bacterial cells were collected from each plate using 1 mL of PBS. Three plates cultured under each condition were pooled and eight serial dilutions (1 : 10 to 1 : 108) were prepared. Each of the eight dilutions (5 μL) was inoculated onto HBA plates in four replicates. The plates were further incubated under Oricon condition for 48 hours at 37°C to determine the CFU numbers.","divisions":[{"label":"Title","span":{"begin":0,"end":133}}],"tracks":[{"project":"MicrobeTaxon","denotations":[{"id":"T71","span":{"begin":20,"end":31},"obj":"199"},{"id":"T72","span":{"begin":506,"end":517},"obj":"199"},{"id":"T73","span":{"begin":1563,"end":1574},"obj":"199"},{"id":"T74","span":{"begin":241,"end":248},"obj":"9606"},{"id":"T75","span":{"begin":461,"end":468},"obj":"9606"},{"id":"T76","span":{"begin":757,"end":766},"obj":"2"},{"id":"T77","span":{"begin":851,"end":860},"obj":"2"},{"id":"T78","span":{"begin":1575,"end":1584},"obj":"2"}],"namespaces":[{"prefix":"_base","uri":"http://purl.bioontology.org/ontology/NCBITAXON/"}],"attributes":[{"subj":"T71","pred":"source","obj":"MicrobeTaxon"},{"subj":"T72","pred":"source","obj":"MicrobeTaxon"},{"subj":"T73","pred":"source","obj":"MicrobeTaxon"},{"subj":"T74","pred":"source","obj":"MicrobeTaxon"},{"subj":"T75","pred":"source","obj":"MicrobeTaxon"},{"subj":"T76","pred":"source","obj":"MicrobeTaxon"},{"subj":"T77","pred":"source","obj":"MicrobeTaxon"},{"subj":"T78","pred":"source","obj":"MicrobeTaxon"}]}],"config":{"attribute types":[{"pred":"source","value type":"selection","values":[{"id":"MicrobeTaxon","color":"#ddec93","default":true}]}]}}