Development of Non-invasive Methods to Study Gut Microbiome - Nutrition - Host Interactions

The aim of this study is to develop and validate an adequate protocol of volatolome (volatile metabolites) measurement in breath of human volunteers (18-65 years) to study gut microbiome - nutrition - host interactions in a general healthy population. For this purpose, breath volatile metabolites (BVM) will be analyzed throughout test days in fasting state and after standardized meals using selected ion flow tube mass spectrometry (SIFT-MS). In addition, gut microbial functions linked notably to hydrogen/methane production, carbohydrates utilization (CAZymes), and in the production of other volatile metabolites of interest (i.e. indole, fatty acid derivatives, ...) will be unraveled through microbiome sequencing data analysis (in silico), metabolomic analyses and measurement of enzymatic activities from saliva, urine and/or fecal samples.

Study Overview

• Status: Recruiting
• Conditions: Healthy
• Intervention / Treatment: Other: Screening visit; Other: Lactulose test; Other: Administration of standardized meals with characterized dietary fiber content

• Study Type: Interventional
• Enrollment (Estimated): 100
• Phase: Not Applicable

Contacts and Locations

• Study Contact: Name: Nathalie Delzenne, Prof.; Phone Number: 0032 2 764 73 69; Email: nathalie.delzenne@uclouvain.be

Study Locations

• Belgium
  • Brabant Wallon
    • Ottignies-Louvain-la-Neuve, Brabant Wallon, Belgium, 1348
      • Not yet recruiting
      • Center of Investigation in Clinical Nutrition (CICN), UCLouvain/CICN
      • Contact: Louise Deldicque, Prof.
  • Brussels
    • Woluwe-Saint-Lambert, Brussels, Belgium, 1200
      • Recruiting
      • Metabolism and Nutrition research group (MNUT), Louvain Drug Research Institute (LDRI), UCLouvain
      • Contact: Nathalie Delzenne, Prof.
    • Woluwe-Saint-Lambert, Brussels, Belgium, 1200
      • Recruiting
      • Testing rooms of Institute of Neuroscience (IONS), UCLouvain/IONS
      • Contact: Louise Deldicque, Prof.

Participation Criteria

Eligibility Criteria

• Ages Eligible for Study: Adult; Older Adult
• Accepts Healthy Volunteers: Yes

Description

Inclusion Criteria:

• Woman or man, aged of 18 to 65 years;
• Body mass index (BMI) between 18 and 30 kg/m2;
• Non-smoker;
• Provision of signed and dated informed consent form;
• Stated willingness to comply with all study procedures and availability for the duration of the study;
• Speaking French.

Exclusion Criteria:

• Subject with severe systemic disease (renal or hepatic failure, cancer, pancreatic cancer, chronic pancreatitis, asthma …)
• Subject with a history or current chronic/severe gastrointestinal disorder such as duodenal ulcer, chronic colitis, or chronic inflammatory disease of the digestive tract disease (Crohn's disease, ulcerative colitis), celiac disease or irritable bowel syndrome;
• Subjects with a history of digestive tract surgery (except appendectomy);
• Subject who had surgery within the two months prior to the study
• Subjects with psychiatric problems and/or using antipsychotics
• Current or recent (< 4 weeks) intake of antibiotics, fiber supplement, and/or any products modulating gut transit (laxative, anti-diarrheal, ...) or gut microbiota composition; these products will also be avoided for the duration of the study;
• Chronic intake of drugs, except contraceptive drug;
• Subject following a low-calorie diet and followed by a doctor or dietician in progress or recently (< 6 weeks);
• Women of childbearing age who are pregnant or breastfeeding or who wish to become pregnant within the next 6 weeks or who are not using an adequate method of contraception (e.g. oral contraception, intrauterine device (IUD), abstinence, ...);
• Subjects who drink more than 3 glasses of alcohol per day (> 30 g of alcohol per day);
• Subjects having participated to another clinical trial two weeks before the screening test visit;
• Subjects who practice an intense sport activity (more than 10 hours per week of intense activity);
• Language barrier, mental or legal incapacity, unwillingness or inability to understand or not being able to participate to the study.

Study Plan

How is the study designed?

Design Details

• Primary Purpose: Other
• Allocation: Non-Randomized
• Interventional Model: Crossover Assignment
• Masking: Single

Number of Arms

5

Arms and Interventions

Participant Group / Arm	Intervention / Treatment
Other: Screening; D-42 to D-7	Other: Screening visit; Subjects will be invited to perform the screening visit to sign the informed consent and then to ensure that they are able to participate in this study.
Other: Lactulose; D0	Other: Lactulose test; Fasted subjects (at least 10 hours) will receive on oral load of 10 g of lactulose. Then, exhaled H2 will be measured every 30 minutes during 4 hours.
Other: Meal 1; D14+/-7	Other: Administration of standardized meals with characterized dietary fiber content; Subjects will receive standardized breakfasts and lunches characterized in terms of dietary fiber content. Urine, saliva, feces samples will be collected. Breath test and gastrointestinal symptoms will be followed during maximum 8h. Subjects will also complete SF36 questionnaire, food diary and stool/transit questionnaire.
Other: Meal 2; D16+/-7	Other: Administration of standardized meals with characterized dietary fiber content; Subjects will receive standardized breakfasts and lunches characterized in terms of dietary fiber content. Urine, saliva, feces samples will be collected. Breath test and gastrointestinal symptoms will be followed during maximum 8h. Subjects will also complete SF36 questionnaire, food diary and stool/transit questionnaire.
Other: Meal 3; D18+/-7	Other: Administration of standardized meals with characterized dietary fiber content; Subjects will receive standardized breakfasts and lunches characterized in terms of dietary fiber content. Urine, saliva, feces samples will be collected. Breath test and gastrointestinal symptoms will be followed during maximum 8h. Subjects will also complete SF36 questionnaire, food diary and stool/transit questionnaire.

What is the study measuring?

Primary Outcome Measures

Outcome Measure	Measure Description	Time Frame
Exhaled volatile compounds	Selected Ion flow tube mass spectrometry (SIFT-MS) - Area under the curve (calculated on 8 hours following breakfast ingestion)	Throughout the entire study, approximately during 24 months

Secondary Outcome Measures

Outcome Measure	Measure Description	Time Frame
Gut microbiota	Illumina sequencing of 16S ribosomal deoxyribonucleic acid (rDNA), quantitative polymerase chain reaction (PCR)	Throughout the entire study, approximately during 24 months
Salivary microbiota	Illumina sequencing of 16SrDNA, quantitative PCR	Throughout the entire study, approximately during 24 months
Gastrointestinal symptom - discomfort	100 mm Visual analog scale (0 = no symptom, 100 = very severe symptom) - Area under the curve (calculated on 8 hours following breakfast ingestion)	Throughout the entire study, approximately during 24 months
Gastrointestinal symptom - nausea	100 mm Visual analog scale (0 = no symptom, 100 = very severe symptom) - Area under the curve (calculated on 8 hours following breakfast ingestion)	Throughout the entire study, approximately during 24 months
Gastrointestinal symptom - bloating	100 mm Visual analog scale (0 = no symptom, 100 = very severe symptom) - Area under the curve (calculated on 8 hours following breakfast ingestion)	Throughout the entire study, approximately during 24 months
Gastrointestinal symptom - flatulences	100 mm Visual analog scale (0 = no symptom, 100 = very severe symptom) - Area under the curve (calculated on 8 hours following breakfast ingestion)	Throughout the entire study, approximately during 24 months
Gastrointestinal symptom - gastrointestinal reflux	100 mm Visual analog scale (0 = no symptom, 100 = very severe symptom) - Area under the curve (calculated on 8 hours following breakfast ingestion)	Throughout the entire study, approximately during 24 months
Gastrointestinal symptom - cramps	100 mm Visual analog scale (0 = no symptom, 100 = very severe symptom) - Area under the curve (calculated on 8 hours following breakfast ingestion)	Throughout the entire study, approximately during 24 months
Gastrointestinal symptom - rumbling	100 mm Visual analog scale (0 = no symptom, 100 = very severe symptom) - Area under the curve (calculated on 8 hours following breakfast ingestion)	Throughout the entire study, approximately during 24 months
Gastrointestinal symptom - burps	100 mm Visual analog scale (0 = no symptom, 100 = very severe symptom)	Throughout the entire study, approximately during 24 months
Gut microbiota metabolic capacity	Enzymatic analysis via in-vitro studies, incubation of fecal extracts	Throughout the entire study, approximately during 24 months
Exhaled hydrogen (H2)	Lactotest 202 - Area under the curve (calculated on 4 hours following lactulose ingestion)	Throughout the entire study, approximately during 24 months
Exhaled methane (CH4)	Lactotest 202 - Area under the curve (calculated on 8 hours following breakfast ingestion)	Throughout the entire study, approximately during 24 months
Exhaled carbon dioxide (CO2)	Lactotest 202 - Area under the curve (calculated on 4 hours following lactulose ingestion)	Throughout the entire study, approximately during 24 months
Carbohydrates utilization - Carbohydrate active enzymes (CAZymes)	Saliva enzymatic activities	Throughout the entire study, approximately during 24 months
Nutrient intake	Food diary (daily log of what the volunteer eats and drinks each day)	Throughout the entire study, approximately during 24 months
Dietary habits	Food frequency questionnaire (FFQ) (record the volunteers' usual diet and dietary habits)	Throughout the entire study, approximately during 24 months
Well-being	The short form (36) health survey (SF36) (0 = worse physical and mental health functioning, 100 = best physical and mental health functioning)	Throughout the entire study, approximately during 24 months
Transit - frequency	Questionnaire on stool frequency (record each defecation time)	Throughout the entire study, approximately during 24 months
Transit - type of stool	Bristol stools scale (BSS) (classify the form of feces into 7 categories; type 1 = severe constipation, type 2 = mild constipation, types 3 and 4 = normal, type 5 = lacking fiber, type 6 = mild diarrhea, type 7 = severe diarrhea)	Throughout the entire study, approximately during 24 months
Transit - ease of passage	5-point Likert's scale (1 = very difficult to defecate, 5 = very easy to defecate)	Throughout the entire study, approximately during 24 months
Transit - level of emergency	5-point Likert's scale (1 = very urgent need to defecate, 5 = very nonurgent need to defecate)	Throughout the entire study, approximately during 24 months
Transit - empty feeling	5-point Likert's scale (1 = very nonempty feeling after defecation, 5 = very empty feeling after defecation)	Throughout the entire study, approximately during 24 months

Collaborators and Investigators

• Sponsor: Université Catholique de Louvain
• Investigators: Principal Investigator: Nathalie Delzenne, Prof., Université Catholique de Louvain

Publications and helpful links

General Publications

• Neyrinck AM, Rodriguez J, Zhang Z, Nazare JA, Bindels LB, Cani PD, Maquet V, Laville M, Bischoff SC, Walter J, Delzenne NM. Breath volatile metabolome reveals the impact of dietary fibres on the gut microbiota: Proof of concept in healthy volunteers. EBioMedicine. 2022 Jun;80:104051. doi: 10.1016/j.ebiom.2022.104051. Epub 2022 May 10.
• Neyrinck AM, Rodriguez J, Zhang Z, Seethaler B, Mailleux F, Vercammen J, Bindels LB, Cani PD, Nazare JA, Maquet V, Laville M, Bischoff SC, Walter J, Delzenne NM. Noninvasive monitoring of fibre fermentation in healthy volunteers by analyzing breath volatile metabolites: lessons from the FiberTAG intervention study. Gut Microbes. 2021 Jan-Dec;13(1):1-16. doi: 10.1080/19490976.2020.1862028.

Study record dates

Study Major Dates

• Study Start (Actual): July 1, 2023
• Primary Completion (Estimated): December 31, 2024
• Study Completion (Estimated): December 31, 2024

Study Registration Dates

• First Submitted: April 17, 2023
• First Submitted That Met QC Criteria: July 13, 2023
• First Posted (Actual): July 18, 2023

Study Record Updates

• Last Update Posted (Actual): July 18, 2023
• Last Update Submitted That Met QC Criteria: July 13, 2023
• Last Verified: July 1, 2023

More Information

Terms related to this study

• Additional Relevant MeSH Terms: Gastrointestinal Agents; Lactulose
• Other Study ID Numbers: MICROBOOST

Plan for Individual participant data (IPD)

• Plan to Share Individual Participant Data (IPD)?: YES

IPD Plan Description

All data will be shared by the partners from the Consortium agreement.

• Gut and salivary microbiota composition;
• Urinary, salivary and fecal metabolites;
• Visual analogue scales data;
• Gut microbial functions;
• Nutrient intake and dietary habits;
• Well-being;
• Transit;
• Concomitant medication;
• Inclusion/exclusion criteria;
• Adverse event.

Data will be shared at the end of the study.

• IPD Sharing Time Frame: At the end of the study
• IPD Sharing Access Criteria: Belonging to the project partner (Consortium agreement)
• IPD Sharing Supporting Information Type: STUDY_PROTOCOL; SAP; CSR

Drug and device information, study documents

• Studies a U.S. FDA-regulated drug product: No
• Studies a U.S. FDA-regulated device product: No