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MB Sample ID: SA121846
| Local Sample ID: | sample1 |
| Subject ID: | SU001507 |
| Subject Type: | Bacteria |
| Subject Species: | Multi-species non-defined biofilm consortium |
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Collection:
| Collection ID: | CO001502 |
| Collection Summary: | Saliva was processed and biofilms were grown using a previously published methodology with modifications (Cleaver, Moazzez and Carpenter, 2019). Saliva samples were centrifuged (5000g for 5 mins), and the supernatant was collected, pooled, boiled in sealed tubes for 20 minutes to sterilise, and left to cool to room temperature; hereafter referred to as sterile saliva. The loose pellet from the spun saliva samples was pooled and combined with enough sterile saliva to facilitate inoculation of the hydroxyapatite (HA) discs. Samples were vortexed vigorously to resuspend the pellet; this constituted the pooled saliva inoculum. The remaining sterile saliva was split into 11 aliquots. D-proline, L-arginine, and glucose were added to the aliquots to achieve concentrations of 10mM, 25mM and 50mM respectively for each (based on concentrations of proline obtained from previous publication (6)). One aliquot was also supplemented with 10mM carbon 13 labelled (C13) D-proline. Two aliquots were left unsupplemented to act as a positive and negative control. Six HA discs in a sterile microtitre plate were inoculated per treatment condition with 1 ml pooled saliva inoculum (negative control discs were inoculated with sterile saliva only to check for sterility) and incubated for 24 hours aerobically in a 40 L aerobic incubator (GenLab Ltd, UK) at 37°C. After this time the discs were washed three times with sterile phosphate buffered saline (PBS) and 1 ml of supplemented/unsupplemented saliva per condition was added to the discs. The discs were then incubated anaerobically in a 3.5 litre anaerobic jar with an Anaerogen anaerobic generator pack (Oxoid, UK) at 37°C for 72 hours. After this spent saliva from the biofilms was removed and stored at -20°C for further analysis, discs were washed three times with PBS, and refreshed with supplemented/unsupplemented saliva and further incubated anaerobically at 37°C for 72 hours. After this final incubation the spent saliva was removed and stored at -20°C for further analysis. |
| Sample Type: | Bacterial cells |
