Impact of Native Chicory Inulin on Change and Persistence of Gut Microbiota

Bifidobacterium counts will be assessed by fluorescence in situ hybridisation - flow cytometry (FISH-FLOW) and 16S rRNA gene sequencing

Total bacteria will be assessed in stool samples by fluorescence in situ hybridisation - flow cytometry.

Microbiota composition at baseline and throughout the course the 13-week intervention will be assessed by 16S rRNA gene sequencing.

Changes in gut fermentation kinetics (rates of change in bacterial counts) will be conducted using 24 h in vitro fermentations at 10 timepoints throughout the 13 week study duration. Aliquots will be collected at 0, 4, 8, 12 and 24 hours. Changes in kinetics will be assessed using fluorescence in situ hybridisation - flow cytometry.

Changes in short-chain fatty acid production will be conducted using 24 h in vitro fermentations at 10 timepoints throughout the 13 week study duration. Aliquots will be collected at 0, 4, 8, 12 and 24 hours during each fermentation and changes in short-chain fatty acid production will be assessed using gas chromatography.

Stool frequency will be assessed as effective number of bowel movements in a daily diary during baseline and both intervention and post-intervention periods.

Stool consistency will be assessed in a daily diary during baseline and both intervention and post intervention periods. Changes will be assessed using the Bristol Stool Form Scale which asks volunteers to rate stool consistency from 1 (being like rabbit droppings - very constipated) up to 7 (being like water with no solid pieces).

Gastrointestinal sensations (bloating) will be assessed in a daily diary during baseline and both intervention and post-intervention on a 100 point visual analogue scale (higher scores indicate higher sensation).

Gastrointestinal sensations (flatulence) will be assessed in a daily diary during baseline and both intervention and post-intervention periods on a 100 point visual analogue scale (higher scores indicate higher sensation).

Gastrointestinal sensations (abdominal pain) will be assessed in a daily diary during baseline and both intervention and post-intervention periods on a 100 point visual analogue scale (higher scores indicate higher sensation).

Gastrointestinal sensations (abdominal pressure) will be assessed in a daily diary during baseline and both intervention and post-intervention periods on a 100 point visual analogue scale (higher scores indicate higher sensation).

Changes in feeling of fullness will be assessed at baseline and throughout both the intervention and post-intervention phases at 9 timepoints in total on a 100 point visual analogue scale.

Changes in feeling of hunger will be assessed in at baseline and throughout both the intervention and post-intervention phases at 9 timepoints in total on a 100 point visual analogue scale.

Changes in desire to eat will be assessed in at baseline and throughout both the intervention and post-intervention phases at 9 timepoints in total on a 100 point visual analogue scale.

Changes in how much could you eat right now will be assessed at baseline and throughout both the intervention and post-intervention phases at 9 timepoints in total on a 100 point visual analogue scale.

Changes in how strong is preoccupation with food iswill be assessed at baseline and throughout both the intervention and post-intervention phases at 9 timepoints in total on a 100 point visual analogue scale.

Changes in feeling of thirst will be assessed at baseline and throughout both the intervention and post-intervention phases at 9 timepoints in total on a 100 point visual analogue scale.

Changes in desire to eat something salty will be assessed at baseline and throughout both the intervention and post-intervention phases at 9 timepoints in total on a 100 point visual analogue scale.

Changes in desire to eat something fatty will be assessed at baseline and throughout both the intervention and post-intervention phases at 9 timepoints in total on a 100 point visual analogue scale.

Changes in desire to eat something sweet will be assessed at baseline and throughout both the intervention and post-intervention phases at 9 timepoints in total on a 100 point visual analogue scale.

Changes in transient mood will be assessed using the Positive and Negative Affect Schedule - Short Form Daily questionnaire. The PANAS possesses 20 self-reported measures of positive affect (PA; 10 items) and negative affect (NA; 10 items) that can be used on multiple occasions. Each volunteer will rate the degree to which they were currently experiencing each item on a 5-point Likert scale. Ratings of positive and negative items were summed to give an overall PA and NA score. Scores range from 10 to 50-higher scores indicate higher levels of PA and NA.

Fecal and blood Metabolites (organic compounds) will be assessed at baseline and throughout both the intervention and post-intervention phases at 9 timepoints in total with Nuclear magnetic resonance (NMR). Unsupervised principal component analysis (PCA) and supervised Orthogonal Projections to Latent Structures Discriminant Analysis (OPLS-DA) analysis will be conducted to assess differences between treatments. Correlation coefficients will be generated to show the strength and direction of changes and differences in metabolites between interventions.