Probiotic Research On Safety, Palatability, Efficacy and Receptivity

Clinical Outcome - Folate, vitamin B12 and B6 concentrations at endline (4-week assessment) compared to baseline

Folate, vitamin B12 and B6 concentrations will be measured using standard clinical lab assay.

Receptivity Outcome

Participant receptivity captured using the receptivity questionnaire at baseline.

Acceptability Outcome

Participant acceptability captured using the acceptability questionnaire at endline (4-week assessment).

Safety Outcome - Reported adverse events at midline (2-week assessment) and endline (4-week assessment)

Reported adverse events, including detailed information related to these adverse events (e.g. frequency, severity, symptoms, timing), will be measured using the bi-weekly study diaries at midline and endline. Proportion of adverse events (AEs) will be measured at midline and endline.

Levels of metabolites (circulating metabolome) at baseline, and endline (4-week assessment)

Functional metabolites in plasma will be measured by Liquid Chromatography-Mass Spectrometry (LC-MS) and Inductively Coupled Plasma-Mass Spectrometry (ICP-MS).

Dietary intake using 24-hour dietary recall at baseline and endline (4-week assessment)

Dietary recalls will be conducted using the Automated Self-Administered 24-hour Dietary Assessment Tool (ASA24) for Canada 2018 (1). This tool measures all food and beverages consumed over a 24-hour period. Dietary recalls will be considered complete if participants provide a minimum of two days of 24-hour recalls.

Dietary intake - exploratory analyses at baseline and endline (4-week assessment)

The investigators will perform exploratory dietary intake analyses, including proportion of participants meeting dietary reference intakes, and dietary patterns of foods.

Dietary intake - HEI and DRI at baseline and endline (4-week assessment)

The investigators will perform exploratory dietary intake analyses, including the proportion of participants meeting the Healthy Eating Index (HEI), and Dietary Inflammatory Index (DRI).

Exploratory: Gut microbial metagenome at baseline and endline (4-week assessment)

Bacterial genomic DNA will be isolated from stool samples and analysed by whole genome sequencing. Exploratory analyses of inferred function of the metagenome will be undertaken using compositional approaches and standardised pipelines to summarise relative taxonomic abundance, diversity and variance within and between groups (2,3). We will further assess metagenome inferred function (mapping reads to KEGG and HUMAnN3), and visualise using compositional and scale-aware approaches (4) (and relate to computationally-derived functions from metagenome data) by measuring functional metabolites (see outcome 7 for additional details).

Exploratory: Gut microbial metabolome at baseline and endline (4-week assessment)

Microbial metabolome (microbial products, short chain fatty acids, vitamins, metals/minerals) will be measured in both groups (5) and data related to computationally-derived functions from metagenome data) using LC-MS and ICP-MS. Data will be processed under standard pipelines with visualisation and pathway analysis by MetaboAnalyst and KEGG databases (6).