Trial Outcomes & Findings for Investigation of the Effects of a Bifidobacterium Breve Strain on Fat Loss in Healthy Adults (NCT NCT04677738)

NCT ID: NCT04677738

Last Updated: 2025-10-17

Results Overview

The difference in change in fat loss from baseline (g), as assessed by Dual-Energy X-Ray Absorptiometry (DXA), between B. breve and placebo after 12 weeks of supplementation. Body tissue density will be measured by a form of X-ray radiation and converted into body fat and muscle mass percentage for assessment

• Recruitment status: COMPLETED
• Study phase: NA
• Target enrollment: 95 participants
• Primary outcome timeframe: 12 weeks from baseline
• Results posted on: 2025-10-17

Participant Flow

Participant milestones

Measure	B. Breve Capsule containing B breve strain. Participants will be instructed to take 2 capsules of placebo for 4 weeks during the run-in period. On day 1 participants will be instructed to take 2 capsules of B. breve strain in the morning before breakfast for 12 weeks. B. breve strain: Probiotic capsule. Participants will be instructed to take 2 capsules of placebo for 4 weeks during the run-in period. On day 1 participants will be instructed to take 2 capsules of B. breve strain in the morning before breakfast for 12 weeks.	Placebo Placebo delivered in capsule format. Participants will be instructed to take 2 capsules of placebo for 4 weeks during the run-in period. On day 1 participants will be instructed to take 2 capsules of placebo in the morning before breakfast for 12 weeks. Placebo: Placebo capsule. Participants will be instructed to take 2 capsules of placebo for 4 weeks during the run-in period. On day 1 participants will be instructed to take 2 capsules of placebo in the morning before breakfast for 12 weeks.
Overall Study STARTED	48	47
Overall Study COMPLETED	44	45
Overall Study NOT COMPLETED	4	2

Reasons for withdrawal

Measure	B. Breve Capsule containing B breve strain. Participants will be instructed to take 2 capsules of placebo for 4 weeks during the run-in period. On day 1 participants will be instructed to take 2 capsules of B. breve strain in the morning before breakfast for 12 weeks. B. breve strain: Probiotic capsule. Participants will be instructed to take 2 capsules of placebo for 4 weeks during the run-in period. On day 1 participants will be instructed to take 2 capsules of B. breve strain in the morning before breakfast for 12 weeks.	Placebo Placebo delivered in capsule format. Participants will be instructed to take 2 capsules of placebo for 4 weeks during the run-in period. On day 1 participants will be instructed to take 2 capsules of placebo in the morning before breakfast for 12 weeks. Placebo: Placebo capsule. Participants will be instructed to take 2 capsules of placebo for 4 weeks during the run-in period. On day 1 participants will be instructed to take 2 capsules of placebo in the morning before breakfast for 12 weeks.
Overall Study Withdrawal by Subject	4	2

Baseline Characteristics

Investigation of the Effects of a Bifidobacterium Breve Strain on Fat Loss in Healthy Adults

Baseline characteristics by cohort

Measure	B. Breve n=48 Participants Capsule containing B breve strain. Participants will be instructed to take 2 capsules of placebo for 4 weeks during the run-in period. On day 1 participants will be instructed to take 2 capsules of B. breve strain in the morning before breakfast for 12 weeks. B. breve strain: Probiotic capsule. Participants will be instructed to take 2 capsules of placebo for 4 weeks during the run-in period. On day 1 participants will be instructed to take 2 capsules of B. breve strain in the morning before breakfast for 12 weeks.	Placebo n=47 Participants Placebo delivered in capsule format. Participants will be instructed to take 2 capsules of placebo for 4 weeks during the run-in period. On day 1 participants will be instructed to take 2 capsules of placebo in the morning before breakfast for 12 weeks. Placebo: Placebo capsule. Participants will be instructed to take 2 capsules of placebo for 4 weeks during the run-in period. On day 1 participants will be instructed to take 2 capsules of placebo in the morning before breakfast for 12 weeks.	Total n=95 Participants Total of all reporting groups
Age, Continuous	41.71 years STANDARD_DEVIATION 10.43 • n=5 Participants	48.04 years STANDARD_DEVIATION 11.16 • n=7 Participants	44.84 years STANDARD_DEVIATION 11.20 • n=5 Participants
Sex: Female, Male Female	38 Participants n=5 Participants	40 Participants n=7 Participants	78 Participants n=5 Participants
Sex: Female, Male Male	10 Participants n=5 Participants	7 Participants n=7 Participants	17 Participants n=5 Participants
Ethnicity (NIH/OMB) Hispanic or Latino	5 Participants n=5 Participants	4 Participants n=7 Participants	9 Participants n=5 Participants
Ethnicity (NIH/OMB) Not Hispanic or Latino	43 Participants n=5 Participants	43 Participants n=7 Participants	86 Participants n=5 Participants
Ethnicity (NIH/OMB) Unknown or Not Reported	0 Participants n=5 Participants	0 Participants n=7 Participants	0 Participants n=5 Participants
Race/Ethnicity, Customized African	1 Participants n=5 Participants	0 Participants n=7 Participants	1 Participants n=5 Participants
Race/Ethnicity, Customized African American	1 Participants n=5 Participants	0 Participants n=7 Participants	1 Participants n=5 Participants
Race/Ethnicity, Customized Central American	1 Participants n=5 Participants	0 Participants n=7 Participants	1 Participants n=5 Participants
Race/Ethnicity, Customized East Asian	3 Participants n=5 Participants	0 Participants n=7 Participants	3 Participants n=5 Participants
Race/Ethnicity, Customized Eastern European White	5 Participants n=5 Participants	4 Participants n=7 Participants	9 Participants n=5 Participants
Race/Ethnicity, Customized Hispanic or Latino	4 Participants n=5 Participants	4 Participants n=7 Participants	8 Participants n=5 Participants
Race/Ethnicity, Customized Middle Eastern	0 Participants n=5 Participants	1 Participants n=7 Participants	1 Participants n=5 Participants
Race/Ethnicity, Customized Mixed	0 Participants n=5 Participants	1 Participants n=7 Participants	1 Participants n=5 Participants
Race/Ethnicity, Customized Other	0 Participants n=5 Participants	1 Participants n=7 Participants	1 Participants n=5 Participants
Race/Ethnicity, Customized South American	1 Participants n=5 Participants	0 Participants n=7 Participants	1 Participants n=5 Participants
Race/Ethnicity, Customized South Asian	3 Participants n=5 Participants	0 Participants n=7 Participants	3 Participants n=5 Participants
Race/Ethnicity, Customized South East Asian	1 Participants n=5 Participants	1 Participants n=7 Participants	2 Participants n=5 Participants
Race/Ethnicity, Customized Western European White	28 Participants n=5 Participants	35 Participants n=7 Participants	63 Participants n=5 Participants
Fat mass	29587 g STANDARD_DEVIATION 5942 • n=5 Participants	30191 g STANDARD_DEVIATION 4645 • n=7 Participants	29886 g STANDARD_DEVIATION 5320 • n=5 Participants
Fat mass ratio	39.4 percentage of body weight STANDARD_DEVIATION 7.5 • n=5 Participants	40.5 percentage of body weight STANDARD_DEVIATION 5.8 • n=7 Participants	40.0 percentage of body weight STANDARD_DEVIATION 6.7 • n=5 Participants

PRIMARY outcome

Timeframe: 12 weeks from baseline

Population: Per-protocol population after excluding the participants with concomitant medication or foods according to the judgement of principal investigator.

The difference in change in fat loss from baseline (g), as assessed by Dual-Energy X-Ray Absorptiometry (DXA), between B. breve and placebo after 12 weeks of supplementation. Body tissue density will be measured by a form of X-ray radiation and converted into body fat and muscle mass percentage for assessment

Outcome measures

Measure	B. Breve n=42 Participants Capsule containing B breve strain. Participants will be instructed to take 2 capsules of placebo for 4 weeks during the run-in period. On day 1 participants will be instructed to take 2 capsules of B. breve strain in the morning before breakfast for 12 weeks. B. breve: Probiotic capsule. Participants will be instructed to take 2 capsules of placebo for 4 weeks during the run-in period. On day 1 participants will be instructed to take 2 capsules of B. breve strain in the morning before breakfast for 12 weeks.	Placebo n=40 Participants Placebo delivered in capsule format. Participants will be instructed to take 2 capsules of placebo for 4 weeks during the run-in period. On day 1 participants will be instructed to take 2 capsules of placebo in the morning before breakfast for 12 weeks. Placebo: Placebo capsule. Participants will be instructed to take 2 capsules of placebo for 4 weeks during the run-in period. On day 1 participants will be instructed to take 2 capsules of placebo in the morning before breakfast for 12 weeks.	Enterotype 2-B. Breve All subjects (regardless of B. breve or Placebo administration) are divided by enterotype. This sub-arm includes the subjects who share the same Enterotype 2, clustered at the genus level.	Enterotype 2-Placebo All subjects (regardless of B. breve or Placebo administration) are divided by enterotype. This sub-arm includes the subjects who share the same Enterotype 2, clustered at the genus level.	Enterotype 3-B. Breve All subjects (regardless of B. breve or Placebo administration) are divided by enterotype. This sub-arm includes the subjects who share the same Enterotype 3, clustered at the genus level.	Enterotype 3-Placebo All subjects (regardless of B. breve or Placebo administration) are divided by enterotype. This sub-arm includes the subjects who share the same Enterotype 3, clustered at the genus level.
Change in Fat Loss (g)	-78.38 g Standard Deviation 2186.66	-9.43 g Standard Deviation 1428.19	—	—	—	—

PRIMARY outcome

Timeframe: 12 weeks from baseline

Population: Per-protocol population after excluding the participants with concomitant medication or foods according to the judgement of principal investigator.

The difference in change in fat loss from baseline (percentage of body weight), as assessed by Dual-Energy X-Ray Absorptiometry (DXA), between B. breve and placebo after 12 weeks of supplementation. Body tissue density will be measured by a form of X-ray radiation and converted into body fat and muscle mass percentage for assessment

Outcome measures

Measure	B. Breve n=42 Participants Capsule containing B breve strain. Participants will be instructed to take 2 capsules of placebo for 4 weeks during the run-in period. On day 1 participants will be instructed to take 2 capsules of B. breve strain in the morning before breakfast for 12 weeks. B. breve: Probiotic capsule. Participants will be instructed to take 2 capsules of placebo for 4 weeks during the run-in period. On day 1 participants will be instructed to take 2 capsules of B. breve strain in the morning before breakfast for 12 weeks.	Placebo n=40 Participants Placebo delivered in capsule format. Participants will be instructed to take 2 capsules of placebo for 4 weeks during the run-in period. On day 1 participants will be instructed to take 2 capsules of placebo in the morning before breakfast for 12 weeks. Placebo: Placebo capsule. Participants will be instructed to take 2 capsules of placebo for 4 weeks during the run-in period. On day 1 participants will be instructed to take 2 capsules of placebo in the morning before breakfast for 12 weeks.	Enterotype 2-B. Breve All subjects (regardless of B. breve or Placebo administration) are divided by enterotype. This sub-arm includes the subjects who share the same Enterotype 2, clustered at the genus level.	Enterotype 2-Placebo All subjects (regardless of B. breve or Placebo administration) are divided by enterotype. This sub-arm includes the subjects who share the same Enterotype 2, clustered at the genus level.	Enterotype 3-B. Breve All subjects (regardless of B. breve or Placebo administration) are divided by enterotype. This sub-arm includes the subjects who share the same Enterotype 3, clustered at the genus level.	Enterotype 3-Placebo All subjects (regardless of B. breve or Placebo administration) are divided by enterotype. This sub-arm includes the subjects who share the same Enterotype 3, clustered at the genus level.
Change in Fat Loss (Percentage of Body Weight)	-0.18 percentage of body weight Standard Deviation 2.01	-0.12 percentage of body weight Standard Deviation 1.35	—	—	—	—

SECONDARY outcome

Timeframe: 12 weeks from baseline

Population: Per-protocol population after excluding the participants with concomitant medication or foods according to the judgement of principal investigator.

The difference in change from baseline between B. breve and placebo in body weight after 12 weeks of supplementation

Outcome measures

Measure	B. Breve n=42 Participants Capsule containing B breve strain. Participants will be instructed to take 2 capsules of placebo for 4 weeks during the run-in period. On day 1 participants will be instructed to take 2 capsules of B. breve strain in the morning before breakfast for 12 weeks. B. breve: Probiotic capsule. Participants will be instructed to take 2 capsules of placebo for 4 weeks during the run-in period. On day 1 participants will be instructed to take 2 capsules of B. breve strain in the morning before breakfast for 12 weeks.	Placebo n=40 Participants Placebo delivered in capsule format. Participants will be instructed to take 2 capsules of placebo for 4 weeks during the run-in period. On day 1 participants will be instructed to take 2 capsules of placebo in the morning before breakfast for 12 weeks. Placebo: Placebo capsule. Participants will be instructed to take 2 capsules of placebo for 4 weeks during the run-in period. On day 1 participants will be instructed to take 2 capsules of placebo in the morning before breakfast for 12 weeks.	Enterotype 2-B. Breve All subjects (regardless of B. breve or Placebo administration) are divided by enterotype. This sub-arm includes the subjects who share the same Enterotype 2, clustered at the genus level.	Enterotype 2-Placebo All subjects (regardless of B. breve or Placebo administration) are divided by enterotype. This sub-arm includes the subjects who share the same Enterotype 2, clustered at the genus level.	Enterotype 3-B. Breve All subjects (regardless of B. breve or Placebo administration) are divided by enterotype. This sub-arm includes the subjects who share the same Enterotype 3, clustered at the genus level.	Enterotype 3-Placebo All subjects (regardless of B. breve or Placebo administration) are divided by enterotype. This sub-arm includes the subjects who share the same Enterotype 3, clustered at the genus level.
Body Weight	-0.04 kg Standard Deviation 2.45	0.10 kg Standard Deviation 1.72	—	—	—	—

SECONDARY outcome

Timeframe: 12 weeks from baseline

Population: Per-protocol population after excluding the participants with concomitant medication or foods according to the judgement of principal investigator.

The difference in change from baseline between B. breve and placebo in BMI after 12 weeks of supplementation

Outcome measures

Measure	B. Breve n=42 Participants Capsule containing B breve strain. Participants will be instructed to take 2 capsules of placebo for 4 weeks during the run-in period. On day 1 participants will be instructed to take 2 capsules of B. breve strain in the morning before breakfast for 12 weeks. B. breve: Probiotic capsule. Participants will be instructed to take 2 capsules of placebo for 4 weeks during the run-in period. On day 1 participants will be instructed to take 2 capsules of B. breve strain in the morning before breakfast for 12 weeks.	Placebo n=40 Participants Placebo delivered in capsule format. Participants will be instructed to take 2 capsules of placebo for 4 weeks during the run-in period. On day 1 participants will be instructed to take 2 capsules of placebo in the morning before breakfast for 12 weeks. Placebo: Placebo capsule. Participants will be instructed to take 2 capsules of placebo for 4 weeks during the run-in period. On day 1 participants will be instructed to take 2 capsules of placebo in the morning before breakfast for 12 weeks.	Enterotype 2-B. Breve All subjects (regardless of B. breve or Placebo administration) are divided by enterotype. This sub-arm includes the subjects who share the same Enterotype 2, clustered at the genus level.	Enterotype 2-Placebo All subjects (regardless of B. breve or Placebo administration) are divided by enterotype. This sub-arm includes the subjects who share the same Enterotype 2, clustered at the genus level.	Enterotype 3-B. Breve All subjects (regardless of B. breve or Placebo administration) are divided by enterotype. This sub-arm includes the subjects who share the same Enterotype 3, clustered at the genus level.	Enterotype 3-Placebo All subjects (regardless of B. breve or Placebo administration) are divided by enterotype. This sub-arm includes the subjects who share the same Enterotype 3, clustered at the genus level.
BMI	-0.02 kilogram per square meter (kg/m^2) Standard Deviation 0.90	0.03 kilogram per square meter (kg/m^2) Standard Deviation 0.65	—	—	—	—

SECONDARY outcome

Timeframe: 12 weeks from baseline

Population: Per-protocol population after excluding the participants with concomitant medication or foods according to the judgement of principal investigator.

The difference in change from baseline between B. breve and placebo in android/gynoid fat ratio as assessed by DXA after 12 weeks of supplementation. Android-gynoid percent fat ratio is a pattern of body fat distribution that is associated with an increased risk for metabolic syndrome in healthy adults.

The measurement of android/gynoid fat ratio is defined as the ratio of the percentage of android fat to the percentage of gynoid fat.

Outcome measures

Measure	B. Breve n=42 Participants Capsule containing B breve strain. Participants will be instructed to take 2 capsules of placebo for 4 weeks during the run-in period. On day 1 participants will be instructed to take 2 capsules of B. breve strain in the morning before breakfast for 12 weeks. B. breve: Probiotic capsule. Participants will be instructed to take 2 capsules of placebo for 4 weeks during the run-in period. On day 1 participants will be instructed to take 2 capsules of B. breve strain in the morning before breakfast for 12 weeks.	Placebo n=40 Participants Placebo delivered in capsule format. Participants will be instructed to take 2 capsules of placebo for 4 weeks during the run-in period. On day 1 participants will be instructed to take 2 capsules of placebo in the morning before breakfast for 12 weeks. Placebo: Placebo capsule. Participants will be instructed to take 2 capsules of placebo for 4 weeks during the run-in period. On day 1 participants will be instructed to take 2 capsules of placebo in the morning before breakfast for 12 weeks.	Enterotype 2-B. Breve All subjects (regardless of B. breve or Placebo administration) are divided by enterotype. This sub-arm includes the subjects who share the same Enterotype 2, clustered at the genus level.	Enterotype 2-Placebo All subjects (regardless of B. breve or Placebo administration) are divided by enterotype. This sub-arm includes the subjects who share the same Enterotype 2, clustered at the genus level.	Enterotype 3-B. Breve All subjects (regardless of B. breve or Placebo administration) are divided by enterotype. This sub-arm includes the subjects who share the same Enterotype 3, clustered at the genus level.	Enterotype 3-Placebo All subjects (regardless of B. breve or Placebo administration) are divided by enterotype. This sub-arm includes the subjects who share the same Enterotype 3, clustered at the genus level.
Android/Gynoid Fat Ratio	-0.02 percentage of gynoid fat ratio Standard Deviation 0.06	0.01 percentage of gynoid fat ratio Standard Deviation 0.06	—	—	—	—

SECONDARY outcome

Timeframe: 12 weeks from baseline

Population: Per-protocol population after excluding the participants with concomitant medication or foods according to the judgement of principal investigator.

The difference in change from baseline between B. breve and placebo in muscle mass (g) as assessed by DXA after 12 weeks of supplementation.

Outcome measures

Measure	B. Breve n=42 Participants Capsule containing B breve strain. Participants will be instructed to take 2 capsules of placebo for 4 weeks during the run-in period. On day 1 participants will be instructed to take 2 capsules of B. breve strain in the morning before breakfast for 12 weeks. B. breve: Probiotic capsule. Participants will be instructed to take 2 capsules of placebo for 4 weeks during the run-in period. On day 1 participants will be instructed to take 2 capsules of B. breve strain in the morning before breakfast for 12 weeks.	Placebo n=40 Participants Placebo delivered in capsule format. Participants will be instructed to take 2 capsules of placebo for 4 weeks during the run-in period. On day 1 participants will be instructed to take 2 capsules of placebo in the morning before breakfast for 12 weeks. Placebo: Placebo capsule. Participants will be instructed to take 2 capsules of placebo for 4 weeks during the run-in period. On day 1 participants will be instructed to take 2 capsules of placebo in the morning before breakfast for 12 weeks.	Enterotype 2-B. Breve All subjects (regardless of B. breve or Placebo administration) are divided by enterotype. This sub-arm includes the subjects who share the same Enterotype 2, clustered at the genus level.	Enterotype 2-Placebo All subjects (regardless of B. breve or Placebo administration) are divided by enterotype. This sub-arm includes the subjects who share the same Enterotype 2, clustered at the genus level.	Enterotype 3-B. Breve All subjects (regardless of B. breve or Placebo administration) are divided by enterotype. This sub-arm includes the subjects who share the same Enterotype 3, clustered at the genus level.	Enterotype 3-Placebo All subjects (regardless of B. breve or Placebo administration) are divided by enterotype. This sub-arm includes the subjects who share the same Enterotype 3, clustered at the genus level.
Muscle Mass (g)	110.64 g Standard Deviation 1380.62	210.82 g Standard Deviation 1157.93	—	—	—	—

SECONDARY outcome

Timeframe: 12 weeks from baseline

Population: Per-protocol population after excluding the participants with concomitant medication or foods according to the judgement of principal investigator.

The difference in change from baseline between B. breve and placebo in muscle mass (percentage of body weight) as assessed by DXA after 12 weeks of supplementation.

Outcome measures

Measure	B. Breve n=42 Participants Capsule containing B breve strain. Participants will be instructed to take 2 capsules of placebo for 4 weeks during the run-in period. On day 1 participants will be instructed to take 2 capsules of B. breve strain in the morning before breakfast for 12 weeks. B. breve: Probiotic capsule. Participants will be instructed to take 2 capsules of placebo for 4 weeks during the run-in period. On day 1 participants will be instructed to take 2 capsules of B. breve strain in the morning before breakfast for 12 weeks.	Placebo n=40 Participants Placebo delivered in capsule format. Participants will be instructed to take 2 capsules of placebo for 4 weeks during the run-in period. On day 1 participants will be instructed to take 2 capsules of placebo in the morning before breakfast for 12 weeks. Placebo: Placebo capsule. Participants will be instructed to take 2 capsules of placebo for 4 weeks during the run-in period. On day 1 participants will be instructed to take 2 capsules of placebo in the morning before breakfast for 12 weeks.	Enterotype 2-B. Breve All subjects (regardless of B. breve or Placebo administration) are divided by enterotype. This sub-arm includes the subjects who share the same Enterotype 2, clustered at the genus level.	Enterotype 2-Placebo All subjects (regardless of B. breve or Placebo administration) are divided by enterotype. This sub-arm includes the subjects who share the same Enterotype 2, clustered at the genus level.	Enterotype 3-B. Breve All subjects (regardless of B. breve or Placebo administration) are divided by enterotype. This sub-arm includes the subjects who share the same Enterotype 3, clustered at the genus level.	Enterotype 3-Placebo All subjects (regardless of B. breve or Placebo administration) are divided by enterotype. This sub-arm includes the subjects who share the same Enterotype 3, clustered at the genus level.
Muscle Mass (Percentage of Body Weight)	0.17 percentage of body weight Standard Deviation 2.04	0.15 percentage of body weight Standard Deviation 1.38	—	—	—	—

SECONDARY outcome

Timeframe: Baseline, 6 weeks and 12 weeks

Population: Per-protocol population after excluding the participants with concomitant medication or foods according to the judgement of principal investigator.

The difference in change from baseline between B. breve and placebo in Waist circumference after 6 and 12 weeks of supplementation.

Outcome measures

Measure	B. Breve n=42 Participants Capsule containing B breve strain. Participants will be instructed to take 2 capsules of placebo for 4 weeks during the run-in period. On day 1 participants will be instructed to take 2 capsules of B. breve strain in the morning before breakfast for 12 weeks. B. breve: Probiotic capsule. Participants will be instructed to take 2 capsules of placebo for 4 weeks during the run-in period. On day 1 participants will be instructed to take 2 capsules of B. breve strain in the morning before breakfast for 12 weeks.	Placebo n=40 Participants Placebo delivered in capsule format. Participants will be instructed to take 2 capsules of placebo for 4 weeks during the run-in period. On day 1 participants will be instructed to take 2 capsules of placebo in the morning before breakfast for 12 weeks. Placebo: Placebo capsule. Participants will be instructed to take 2 capsules of placebo for 4 weeks during the run-in period. On day 1 participants will be instructed to take 2 capsules of placebo in the morning before breakfast for 12 weeks.	Enterotype 2-B. Breve All subjects (regardless of B. breve or Placebo administration) are divided by enterotype. This sub-arm includes the subjects who share the same Enterotype 2, clustered at the genus level.	Enterotype 2-Placebo All subjects (regardless of B. breve or Placebo administration) are divided by enterotype. This sub-arm includes the subjects who share the same Enterotype 2, clustered at the genus level.	Enterotype 3-B. Breve All subjects (regardless of B. breve or Placebo administration) are divided by enterotype. This sub-arm includes the subjects who share the same Enterotype 3, clustered at the genus level.	Enterotype 3-Placebo All subjects (regardless of B. breve or Placebo administration) are divided by enterotype. This sub-arm includes the subjects who share the same Enterotype 3, clustered at the genus level.
Waist Circumference 6 weeks from baseline	-0.22 cm Standard Deviation 3.32	-0.54 cm Standard Deviation 3.31	—	—	—	—
Waist Circumference 12 weeks from baseline	-0.06 cm Standard Deviation 2.92	-0.54 cm Standard Deviation 3.35	—	—	—	—

SECONDARY outcome

Timeframe: Baseline, 6 weeks and 12 weeks

Population: Per-protocol population after excluding the participants with concomitant medication or foods according to the judgement of principal investigator.

The difference in change from baseline between B. breve and placebo in hip circumference after 6 and 12 weeks of supplementation.

Outcome measures

Measure	B. Breve n=42 Participants Capsule containing B breve strain. Participants will be instructed to take 2 capsules of placebo for 4 weeks during the run-in period. On day 1 participants will be instructed to take 2 capsules of B. breve strain in the morning before breakfast for 12 weeks. B. breve: Probiotic capsule. Participants will be instructed to take 2 capsules of placebo for 4 weeks during the run-in period. On day 1 participants will be instructed to take 2 capsules of B. breve strain in the morning before breakfast for 12 weeks.	Placebo n=40 Participants Placebo delivered in capsule format. Participants will be instructed to take 2 capsules of placebo for 4 weeks during the run-in period. On day 1 participants will be instructed to take 2 capsules of placebo in the morning before breakfast for 12 weeks. Placebo: Placebo capsule. Participants will be instructed to take 2 capsules of placebo for 4 weeks during the run-in period. On day 1 participants will be instructed to take 2 capsules of placebo in the morning before breakfast for 12 weeks.	Enterotype 2-B. Breve All subjects (regardless of B. breve or Placebo administration) are divided by enterotype. This sub-arm includes the subjects who share the same Enterotype 2, clustered at the genus level.	Enterotype 2-Placebo All subjects (regardless of B. breve or Placebo administration) are divided by enterotype. This sub-arm includes the subjects who share the same Enterotype 2, clustered at the genus level.	Enterotype 3-B. Breve All subjects (regardless of B. breve or Placebo administration) are divided by enterotype. This sub-arm includes the subjects who share the same Enterotype 3, clustered at the genus level.	Enterotype 3-Placebo All subjects (regardless of B. breve or Placebo administration) are divided by enterotype. This sub-arm includes the subjects who share the same Enterotype 3, clustered at the genus level.
Hip Circumference 6 weeks from baseline	-0.67 cm Standard Deviation 3.97	-0.80 cm Standard Deviation 3.77	—	—	—	—
Hip Circumference 12 weeks from baseline	-1.04 cm Standard Deviation 3.58	-0.97 cm Standard Deviation 4.51	—	—	—	—

SECONDARY outcome

Timeframe: Baseline, 6 weeks and 12 weeks

Population: Per-protocol population after excluding the participants with concomitant medication or foods according to the judgement of principal investigator.

The difference in change from baseline between B. breve and placebo in waist/hip circumference ratio after 6 and 12 weeks of supplementation.

The waist/hip circumference ratio is defined as the percentage of waist circumference to hip circumference. The change in waist/hip circumference ratio from baseline to week 12 was calculated.

Outcome measures

Measure	B. Breve n=42 Participants Capsule containing B breve strain. Participants will be instructed to take 2 capsules of placebo for 4 weeks during the run-in period. On day 1 participants will be instructed to take 2 capsules of B. breve strain in the morning before breakfast for 12 weeks. B. breve: Probiotic capsule. Participants will be instructed to take 2 capsules of placebo for 4 weeks during the run-in period. On day 1 participants will be instructed to take 2 capsules of B. breve strain in the morning before breakfast for 12 weeks.	Placebo n=40 Participants Placebo delivered in capsule format. Participants will be instructed to take 2 capsules of placebo for 4 weeks during the run-in period. On day 1 participants will be instructed to take 2 capsules of placebo in the morning before breakfast for 12 weeks. Placebo: Placebo capsule. Participants will be instructed to take 2 capsules of placebo for 4 weeks during the run-in period. On day 1 participants will be instructed to take 2 capsules of placebo in the morning before breakfast for 12 weeks.	Enterotype 2-B. Breve All subjects (regardless of B. breve or Placebo administration) are divided by enterotype. This sub-arm includes the subjects who share the same Enterotype 2, clustered at the genus level.	Enterotype 2-Placebo All subjects (regardless of B. breve or Placebo administration) are divided by enterotype. This sub-arm includes the subjects who share the same Enterotype 2, clustered at the genus level.	Enterotype 3-B. Breve All subjects (regardless of B. breve or Placebo administration) are divided by enterotype. This sub-arm includes the subjects who share the same Enterotype 3, clustered at the genus level.	Enterotype 3-Placebo All subjects (regardless of B. breve or Placebo administration) are divided by enterotype. This sub-arm includes the subjects who share the same Enterotype 3, clustered at the genus level.
Waist/Hip Circumference Ratio 6 weeks from baseline	0.00 percentage of hip circumference Standard Deviation 0.04	0.00 percentage of hip circumference Standard Deviation 0.05	—	—	—	—
Waist/Hip Circumference Ratio 12 weeks from baseline	0.01 percentage of hip circumference Standard Deviation 0.04	0.00 percentage of hip circumference Standard Deviation 0.05	—	—	—	—

SECONDARY outcome

Timeframe: 12 weeks from baseline

Population: Per-protocol population after excluding the participants with concomitant medication or foods according to the judgement of principal investigator.

Alpha diversity at baseline and after 12-week intake were calculated by R. Alpha diversity index is usually unitless since it is a numerical value calculated by a mathematical formula, not a physical quantity.

The Shannon Index (or Shannon-Wiener/Shannon-Weaver index) is a widely used metric in ecology to quantify alpha diversity, which measures both species richness and evenness.

The value of the Shannon Index ranges from a minimum of zero, when there is only one species present (no diversity), to a maximum of ln(S), the natural logarithm of the total number of species (richness) to the base of Euler's number (e), which means all species are equally abundant. High values indicate a diverse, balanced community.

Outcome measures

Measure	B. Breve n=42 Participants Capsule containing B breve strain. Participants will be instructed to take 2 capsules of placebo for 4 weeks during the run-in period. On day 1 participants will be instructed to take 2 capsules of B. breve strain in the morning before breakfast for 12 weeks. B. breve: Probiotic capsule. Participants will be instructed to take 2 capsules of placebo for 4 weeks during the run-in period. On day 1 participants will be instructed to take 2 capsules of B. breve strain in the morning before breakfast for 12 weeks.	Placebo n=40 Participants Placebo delivered in capsule format. Participants will be instructed to take 2 capsules of placebo for 4 weeks during the run-in period. On day 1 participants will be instructed to take 2 capsules of placebo in the morning before breakfast for 12 weeks. Placebo: Placebo capsule. Participants will be instructed to take 2 capsules of placebo for 4 weeks during the run-in period. On day 1 participants will be instructed to take 2 capsules of placebo in the morning before breakfast for 12 weeks.	Enterotype 2-B. Breve All subjects (regardless of B. breve or Placebo administration) are divided by enterotype. This sub-arm includes the subjects who share the same Enterotype 2, clustered at the genus level.	Enterotype 2-Placebo All subjects (regardless of B. breve or Placebo administration) are divided by enterotype. This sub-arm includes the subjects who share the same Enterotype 2, clustered at the genus level.	Enterotype 3-B. Breve All subjects (regardless of B. breve or Placebo administration) are divided by enterotype. This sub-arm includes the subjects who share the same Enterotype 3, clustered at the genus level.	Enterotype 3-Placebo All subjects (regardless of B. breve or Placebo administration) are divided by enterotype. This sub-arm includes the subjects who share the same Enterotype 3, clustered at the genus level.
Microbiota Analysis: Shannon Index Shannon@baseline	5.92 Shannon Index Standard Deviation 0.42	5.88 Shannon Index Standard Deviation 0.52	—	—	—	—
Microbiota Analysis: Shannon Index Shannon@12 weeks	6.00 Shannon Index Standard Deviation 0.43	5.97 Shannon Index Standard Deviation 0.50	—	—	—	—

SECONDARY outcome

Timeframe: 12 weeks from baseline

Population: Per-protocol population after excluding the participants with concomitant medication or foods according to the judgement of principal investigator.

Alpha diversity at baseline and after 12-week intake were calculated by R. Alpha diversity index is usually unitless since it is a numerical value calculated by a mathematical formula, not a physical quantity.

The Chao1 Index estimates total species richness, including rare/undetected species.

The value of the Chao1 Index ranges from a minimum, close to zero (suggesting reduced richness, often observed in dysbiosis) to a theoretically unbounded maximum, as it depends on rare species. Higher values indicate greater estimated species richness, generally associated with a healthier gut microbiome, linked to resilience, metabolic versatility, and protection against pathogens.

Outcome measures

Measure	B. Breve n=42 Participants Capsule containing B breve strain. Participants will be instructed to take 2 capsules of placebo for 4 weeks during the run-in period. On day 1 participants will be instructed to take 2 capsules of B. breve strain in the morning before breakfast for 12 weeks. B. breve: Probiotic capsule. Participants will be instructed to take 2 capsules of placebo for 4 weeks during the run-in period. On day 1 participants will be instructed to take 2 capsules of B. breve strain in the morning before breakfast for 12 weeks.	Placebo n=40 Participants Placebo delivered in capsule format. Participants will be instructed to take 2 capsules of placebo for 4 weeks during the run-in period. On day 1 participants will be instructed to take 2 capsules of placebo in the morning before breakfast for 12 weeks. Placebo: Placebo capsule. Participants will be instructed to take 2 capsules of placebo for 4 weeks during the run-in period. On day 1 participants will be instructed to take 2 capsules of placebo in the morning before breakfast for 12 weeks.	Enterotype 2-B. Breve All subjects (regardless of B. breve or Placebo administration) are divided by enterotype. This sub-arm includes the subjects who share the same Enterotype 2, clustered at the genus level.	Enterotype 2-Placebo All subjects (regardless of B. breve or Placebo administration) are divided by enterotype. This sub-arm includes the subjects who share the same Enterotype 2, clustered at the genus level.	Enterotype 3-B. Breve All subjects (regardless of B. breve or Placebo administration) are divided by enterotype. This sub-arm includes the subjects who share the same Enterotype 3, clustered at the genus level.	Enterotype 3-Placebo All subjects (regardless of B. breve or Placebo administration) are divided by enterotype. This sub-arm includes the subjects who share the same Enterotype 3, clustered at the genus level.
Microbiota Analysis: Chao1 Index Chao1@12 weeks	165.90 Chao1 index Standard Deviation 40.57	158.20 Chao1 index Standard Deviation 39.36	—	—	—	—
Microbiota Analysis: Chao1 Index Chao1@baseline	135.90 Chao1 index Standard Deviation 31.16	141.50 Chao1 index Standard Deviation 36.73	—	—	—	—

SECONDARY outcome

Timeframe: 12 weeks from baseline

Population: Per-protocol population after excluding the participants with concomitant medication or foods according to the judgement of principal investigator.

Alpha diversity at baseline and after 12-week intake were calculated by R. Alpha diversity index is usually unitless since it is a numerical value calculated by a mathematical formula, not a physical quantity.

The Evenness Index measures how evenly individuals are distributed among species in a community, such as the gut microbiome. It complements species richness (the number of species) to describe diversity, ranging from 0 (only a single species, meaning there is no evenness.) to 1, where 1 indicates perfect evenness, meaning all species are equally abundant.

Higher Evenness Index value (closer to 1) is generally considered better, as it suggests a more balanced and stable ecosystem. Conversely, lower value (closer to 0) is generally considered worse, as it suggests that a few species dominate the community, which can be associated with dysbiosis or an unhealthy gut microbiome.

Outcome measures

Measure	B. Breve n=42 Participants Capsule containing B breve strain. Participants will be instructed to take 2 capsules of placebo for 4 weeks during the run-in period. On day 1 participants will be instructed to take 2 capsules of B. breve strain in the morning before breakfast for 12 weeks. B. breve: Probiotic capsule. Participants will be instructed to take 2 capsules of placebo for 4 weeks during the run-in period. On day 1 participants will be instructed to take 2 capsules of B. breve strain in the morning before breakfast for 12 weeks.	Placebo n=40 Participants Placebo delivered in capsule format. Participants will be instructed to take 2 capsules of placebo for 4 weeks during the run-in period. On day 1 participants will be instructed to take 2 capsules of placebo in the morning before breakfast for 12 weeks. Placebo: Placebo capsule. Participants will be instructed to take 2 capsules of placebo for 4 weeks during the run-in period. On day 1 participants will be instructed to take 2 capsules of placebo in the morning before breakfast for 12 weeks.	Enterotype 2-B. Breve All subjects (regardless of B. breve or Placebo administration) are divided by enterotype. This sub-arm includes the subjects who share the same Enterotype 2, clustered at the genus level.	Enterotype 2-Placebo All subjects (regardless of B. breve or Placebo administration) are divided by enterotype. This sub-arm includes the subjects who share the same Enterotype 2, clustered at the genus level.	Enterotype 3-B. Breve All subjects (regardless of B. breve or Placebo administration) are divided by enterotype. This sub-arm includes the subjects who share the same Enterotype 3, clustered at the genus level.	Enterotype 3-Placebo All subjects (regardless of B. breve or Placebo administration) are divided by enterotype. This sub-arm includes the subjects who share the same Enterotype 3, clustered at the genus level.
Microbiota Analysis: Evenness Index Evenness@12 weeks	0.82 Evenness Index Standard Deviation 0.030	0.82 Evenness Index Standard Deviation 0.0058	—	—	—	—
Microbiota Analysis: Evenness Index Evenness@baseline	0.84 Evenness Index Standard Deviation 0.026	0.84 Evenness Index Standard Deviation 0.038	—	—	—	—

SECONDARY outcome

Timeframe: 12 weeks from baseline

Population: Per-protocol population after excluding the participants with concomitant medication or foods according to the judgement of principal investigator.

Alpha diversity at baseline and after 12-week intake were calculated by R. Alpha diversity index is usually unitless since it is a numerical value calculated by a mathematical formula, not a physical quantity.

Observed ASV (Amplicon Sequence Variants) Index refers to the number of distinct Amplicon Sequence Variants (ASVs) detected in a sample. ASVs are highly resolved sequences used to identify and differentiate microbial taxa, providing finer resolution than traditional Operational Taxonomic Units (OTUs).

The range of Observed ASV Index in gut microbiome samples can range from a few hundred to several thousand, depending on several factors, including sample source, health status of the host, diet and lifestyle, as well as sequencing depth and methodology. Typically, higher Observed ASV Index indicates a more diverse microbial community, which can contribute to improved gut function and overall health.

Outcome measures

Measure	B. Breve n=42 Participants Capsule containing B breve strain. Participants will be instructed to take 2 capsules of placebo for 4 weeks during the run-in period. On day 1 participants will be instructed to take 2 capsules of B. breve strain in the morning before breakfast for 12 weeks. B. breve: Probiotic capsule. Participants will be instructed to take 2 capsules of placebo for 4 weeks during the run-in period. On day 1 participants will be instructed to take 2 capsules of B. breve strain in the morning before breakfast for 12 weeks.	Placebo n=40 Participants Placebo delivered in capsule format. Participants will be instructed to take 2 capsules of placebo for 4 weeks during the run-in period. On day 1 participants will be instructed to take 2 capsules of placebo in the morning before breakfast for 12 weeks. Placebo: Placebo capsule. Participants will be instructed to take 2 capsules of placebo for 4 weeks during the run-in period. On day 1 participants will be instructed to take 2 capsules of placebo in the morning before breakfast for 12 weeks.	Enterotype 2-B. Breve All subjects (regardless of B. breve or Placebo administration) are divided by enterotype. This sub-arm includes the subjects who share the same Enterotype 2, clustered at the genus level.	Enterotype 2-Placebo All subjects (regardless of B. breve or Placebo administration) are divided by enterotype. This sub-arm includes the subjects who share the same Enterotype 2, clustered at the genus level.	Enterotype 3-B. Breve All subjects (regardless of B. breve or Placebo administration) are divided by enterotype. This sub-arm includes the subjects who share the same Enterotype 3, clustered at the genus level.	Enterotype 3-Placebo All subjects (regardless of B. breve or Placebo administration) are divided by enterotype. This sub-arm includes the subjects who share the same Enterotype 3, clustered at the genus level.
Microbiota Analysis: Observed ASV Index Observed ASV@baseline	136.60 Observed ASV Index Standard Deviation 32.94	132.40 Observed ASV Index Standard Deviation 35.65	—	—	—	—
Microbiota Analysis: Observed ASV Index Observed ASV@12 weeks	163.00 Observed ASV Index Standard Deviation 38.68	156.1 Observed ASV Index Standard Deviation 38.3	—	—	—	—

SECONDARY outcome

Timeframe: 12 weeks from baseline

Population: Per-protocol population after excluding the participants with concomitant medication or foods according to the judgement of principal investigator.

Alpha diversity at baseline and after 12-week intake were calculated by R. Alpha diversity index is usually unitless since it is a numerical value calculated by a mathematical formula, not a physical quantity.

Faith's Phylogenetic Diversity (PD) is a measure used in ecology to quantify the biodiversity of a sample based on the phylogenetic tree. In the context of the gut microbiome, it specifically measures the diversity of microbial species by considering both the number of species present (richness) and the phylogenetic differences between them.

In practice, the values can range from very low (close to zero, indicating low microbial diversity linked to various health issues) to very high (indicates greater evolutionary diversity, which may enhance ecosystem resilience and is generally considered better in the context of gut microbiome health.).

Outcome measures

Measure	B. Breve n=42 Participants Capsule containing B breve strain. Participants will be instructed to take 2 capsules of placebo for 4 weeks during the run-in period. On day 1 participants will be instructed to take 2 capsules of B. breve strain in the morning before breakfast for 12 weeks. B. breve: Probiotic capsule. Participants will be instructed to take 2 capsules of placebo for 4 weeks during the run-in period. On day 1 participants will be instructed to take 2 capsules of B. breve strain in the morning before breakfast for 12 weeks.	Placebo n=40 Participants Placebo delivered in capsule format. Participants will be instructed to take 2 capsules of placebo for 4 weeks during the run-in period. On day 1 participants will be instructed to take 2 capsules of placebo in the morning before breakfast for 12 weeks. Placebo: Placebo capsule. Participants will be instructed to take 2 capsules of placebo for 4 weeks during the run-in period. On day 1 participants will be instructed to take 2 capsules of placebo in the morning before breakfast for 12 weeks.	Enterotype 2-B. Breve All subjects (regardless of B. breve or Placebo administration) are divided by enterotype. This sub-arm includes the subjects who share the same Enterotype 2, clustered at the genus level.	Enterotype 2-Placebo All subjects (regardless of B. breve or Placebo administration) are divided by enterotype. This sub-arm includes the subjects who share the same Enterotype 2, clustered at the genus level.	Enterotype 3-B. Breve All subjects (regardless of B. breve or Placebo administration) are divided by enterotype. This sub-arm includes the subjects who share the same Enterotype 3, clustered at the genus level.	Enterotype 3-Placebo All subjects (regardless of B. breve or Placebo administration) are divided by enterotype. This sub-arm includes the subjects who share the same Enterotype 3, clustered at the genus level.
Microbiota Analysis: Faith's PD Index Faith's PD@baseline	14.82 Faith's PD Index Standard Deviation 3.17	14.57 Faith's PD Index Standard Deviation 3.84	—	—	—	—
Microbiota Analysis: Faith's PD Index Faith's PD@12 weeks	15.71 Faith's PD Index Standard Deviation 3.81	15.53 Faith's PD Index Standard Deviation 3.49	—	—	—	—

SECONDARY outcome

Timeframe: 12 weeks from baseline

Population: Per-protocol population after excluding the participants with concomitant medication or foods according to the judgement of principal investigator.

The change from baseline between B. breve and placebo in weight in participant groups classified by microbiota composition at week 0, after 12 weeks of supplementation. These individual outcomes comprise relevant aspects of body composition. Each outcome will be analyzed separately and interpreted separately as well as collectively to inform relevant changes to body composition as a whole during the study.

Outcome measures

Measure	B. Breve n=25 Participants Capsule containing B breve strain. Participants will be instructed to take 2 capsules of placebo for 4 weeks during the run-in period. On day 1 participants will be instructed to take 2 capsules of B. breve strain in the morning before breakfast for 12 weeks. B. breve: Probiotic capsule. Participants will be instructed to take 2 capsules of placebo for 4 weeks during the run-in period. On day 1 participants will be instructed to take 2 capsules of B. breve strain in the morning before breakfast for 12 weeks.	Placebo n=22 Participants Placebo delivered in capsule format. Participants will be instructed to take 2 capsules of placebo for 4 weeks during the run-in period. On day 1 participants will be instructed to take 2 capsules of placebo in the morning before breakfast for 12 weeks. Placebo: Placebo capsule. Participants will be instructed to take 2 capsules of placebo for 4 weeks during the run-in period. On day 1 participants will be instructed to take 2 capsules of placebo in the morning before breakfast for 12 weeks.	Enterotype 2-B. Breve n=10 Participants All subjects (regardless of B. breve or Placebo administration) are divided by enterotype. This sub-arm includes the subjects who share the same Enterotype 2, clustered at the genus level.	Enterotype 2-Placebo n=15 Participants All subjects (regardless of B. breve or Placebo administration) are divided by enterotype. This sub-arm includes the subjects who share the same Enterotype 2, clustered at the genus level.	Enterotype 3-B. Breve n=7 Participants All subjects (regardless of B. breve or Placebo administration) are divided by enterotype. This sub-arm includes the subjects who share the same Enterotype 3, clustered at the genus level.	Enterotype 3-Placebo n=3 Participants All subjects (regardless of B. breve or Placebo administration) are divided by enterotype. This sub-arm includes the subjects who share the same Enterotype 3, clustered at the genus level.
Change in Weight in Participant Groups Classified by Microbiota Composition at Week 0	-0.17 kg Standard Deviation 2.76	0.46 kg Standard Deviation 1.79	0.58 kg Standard Deviation 1.96	-0.25 kg Standard Deviation 1.37	-0.44 kg Standard Deviation 1.93	-0.77 kg Standard Deviation 2.63

SECONDARY outcome

Timeframe: 12 weeks from baseline

Population: Per-protocol population after excluding the participants with concomitant medication or foods according to the judgement of principal investigator.

The change from baseline between B. breve and placebo in body mass index in participant groups classified by microbiota composition at week 0, after 12 weeks of supplementation. These individual outcomes comprise relevant aspects of body composition. Each outcome will be analyzed separately and interpreted separately as well as collectively to inform relevant changes to body composition as a whole during the study.

Outcome measures

Measure	B. Breve n=25 Participants Capsule containing B breve strain. Participants will be instructed to take 2 capsules of placebo for 4 weeks during the run-in period. On day 1 participants will be instructed to take 2 capsules of B. breve strain in the morning before breakfast for 12 weeks. B. breve: Probiotic capsule. Participants will be instructed to take 2 capsules of placebo for 4 weeks during the run-in period. On day 1 participants will be instructed to take 2 capsules of B. breve strain in the morning before breakfast for 12 weeks.	Placebo n=22 Participants Placebo delivered in capsule format. Participants will be instructed to take 2 capsules of placebo for 4 weeks during the run-in period. On day 1 participants will be instructed to take 2 capsules of placebo in the morning before breakfast for 12 weeks. Placebo: Placebo capsule. Participants will be instructed to take 2 capsules of placebo for 4 weeks during the run-in period. On day 1 participants will be instructed to take 2 capsules of placebo in the morning before breakfast for 12 weeks.	Enterotype 2-B. Breve n=10 Participants All subjects (regardless of B. breve or Placebo administration) are divided by enterotype. This sub-arm includes the subjects who share the same Enterotype 2, clustered at the genus level.	Enterotype 2-Placebo n=15 Participants All subjects (regardless of B. breve or Placebo administration) are divided by enterotype. This sub-arm includes the subjects who share the same Enterotype 2, clustered at the genus level.	Enterotype 3-B. Breve n=7 Participants All subjects (regardless of B. breve or Placebo administration) are divided by enterotype. This sub-arm includes the subjects who share the same Enterotype 3, clustered at the genus level.	Enterotype 3-Placebo n=3 Participants All subjects (regardless of B. breve or Placebo administration) are divided by enterotype. This sub-arm includes the subjects who share the same Enterotype 3, clustered at the genus level.
Change in Body Mass Index in Participant Groups Classified by Microbiota Composition at Week 0	-0.06 kilogram per square meter (kg/m^2) Standard Deviation 1.04	0.16 kilogram per square meter (kg/m^2) Standard Deviation 0.67	0.18 kilogram per square meter (kg/m^2) Standard Deviation 0.6	-0.11 kilogram per square meter (kg/m^2) Standard Deviation 0.52	-0.13 kilogram per square meter (kg/m^2) Standard Deviation 0.7	-0.3 kilogram per square meter (kg/m^2) Standard Deviation 1.01

SECONDARY outcome

Timeframe: 12 weeks from baseline

Population: Per-protocol population after excluding the participants with concomitant medication or foods according to the judgement of principal investigator.

The change from baseline between B. breve and placebo in total body fat (g) in participant groups classified by microbiota composition at week 0, after 12 weeks of supplementation. These individual outcomes comprise relevant aspects of body composition. Each outcome will be analyzed separately and interpreted separately as well as collectively to inform relevant changes to body composition as a whole during the study.

Outcome measures

Measure	B. Breve n=25 Participants Capsule containing B breve strain. Participants will be instructed to take 2 capsules of placebo for 4 weeks during the run-in period. On day 1 participants will be instructed to take 2 capsules of B. breve strain in the morning before breakfast for 12 weeks. B. breve: Probiotic capsule. Participants will be instructed to take 2 capsules of placebo for 4 weeks during the run-in period. On day 1 participants will be instructed to take 2 capsules of B. breve strain in the morning before breakfast for 12 weeks.	Placebo n=22 Participants Placebo delivered in capsule format. Participants will be instructed to take 2 capsules of placebo for 4 weeks during the run-in period. On day 1 participants will be instructed to take 2 capsules of placebo in the morning before breakfast for 12 weeks. Placebo: Placebo capsule. Participants will be instructed to take 2 capsules of placebo for 4 weeks during the run-in period. On day 1 participants will be instructed to take 2 capsules of placebo in the morning before breakfast for 12 weeks.	Enterotype 2-B. Breve n=10 Participants All subjects (regardless of B. breve or Placebo administration) are divided by enterotype. This sub-arm includes the subjects who share the same Enterotype 2, clustered at the genus level.	Enterotype 2-Placebo n=15 Participants All subjects (regardless of B. breve or Placebo administration) are divided by enterotype. This sub-arm includes the subjects who share the same Enterotype 2, clustered at the genus level.	Enterotype 3-B. Breve n=7 Participants All subjects (regardless of B. breve or Placebo administration) are divided by enterotype. This sub-arm includes the subjects who share the same Enterotype 3, clustered at the genus level.	Enterotype 3-Placebo n=3 Participants All subjects (regardless of B. breve or Placebo administration) are divided by enterotype. This sub-arm includes the subjects who share the same Enterotype 3, clustered at the genus level.
Change in Total Body Fat (g) in Participant Groups Classified by Microbiota Composition at Week 0	-238.24 g Standard Deviation 2301.91	55.77 g Standard Deviation 1290.03	31.8 g Standard Deviation 2251.64	51.67 g Standard Deviation 1506.94	335.14 g Standard Deviation 1883.4	-793 g Standard Deviation 2325.77

SECONDARY outcome

Timeframe: 12 weeks from baseline

Population: Per-protocol population after excluding the participants with concomitant medication or foods according to the judgement of principal investigator.

The change from baseline between B. breve and placebo in total body fat (%) in participant groups classified by microbiota composition at week 0, after 12 weeks of supplementation. These individual outcomes comprise relevant aspects of body composition. Each outcome will be analyzed separately and interpreted separately as well as collectively to inform relevant changes to body composition as a whole during the study.

Outcome measures

Measure	B. Breve n=25 Participants Capsule containing B breve strain. Participants will be instructed to take 2 capsules of placebo for 4 weeks during the run-in period. On day 1 participants will be instructed to take 2 capsules of B. breve strain in the morning before breakfast for 12 weeks. B. breve: Probiotic capsule. Participants will be instructed to take 2 capsules of placebo for 4 weeks during the run-in period. On day 1 participants will be instructed to take 2 capsules of B. breve strain in the morning before breakfast for 12 weeks.	Placebo n=22 Participants Placebo delivered in capsule format. Participants will be instructed to take 2 capsules of placebo for 4 weeks during the run-in period. On day 1 participants will be instructed to take 2 capsules of placebo in the morning before breakfast for 12 weeks. Placebo: Placebo capsule. Participants will be instructed to take 2 capsules of placebo for 4 weeks during the run-in period. On day 1 participants will be instructed to take 2 capsules of placebo in the morning before breakfast for 12 weeks.	Enterotype 2-B. Breve n=10 Participants All subjects (regardless of B. breve or Placebo administration) are divided by enterotype. This sub-arm includes the subjects who share the same Enterotype 2, clustered at the genus level.	Enterotype 2-Placebo n=15 Participants All subjects (regardless of B. breve or Placebo administration) are divided by enterotype. This sub-arm includes the subjects who share the same Enterotype 2, clustered at the genus level.	Enterotype 3-B. Breve n=7 Participants All subjects (regardless of B. breve or Placebo administration) are divided by enterotype. This sub-arm includes the subjects who share the same Enterotype 3, clustered at the genus level.	Enterotype 3-Placebo n=3 Participants All subjects (regardless of B. breve or Placebo administration) are divided by enterotype. This sub-arm includes the subjects who share the same Enterotype 3, clustered at the genus level.
Change in Total Body Fat (% of Body Weight) in Participant Groups Classified by Microbiota Composition at Week 0	-0.28 percentage of body weight Standard Deviation 2.02	-0.14 percentage of body weight Standard Deviation 1.05	-0.31 percentage of body weight Standard Deviation 2.26	0.09 percentage of body weight Standard Deviation 1.64	0.38 percentage of body weight Standard Deviation 1.79	-1.02 percentage of body weight Standard Deviation 1.95

SECONDARY outcome

Timeframe: 12 weeks from baseline

Population: Per-protocol population after excluding the participants with concomitant medication or foods according to the judgement of principal investigator.

The change from baseline between B. breve and placebo in muscle mass (g) in participant groups classified by microbiota composition at week 0, after 12 weeks of supplementation. These individual outcomes comprise relevant aspects of body composition. Each outcome will be analyzed separately and interpreted separately as well as collectively to inform relevant changes to body composition as a whole during the study.

Outcome measures

Measure	B. Breve n=25 Participants Capsule containing B breve strain. Participants will be instructed to take 2 capsules of placebo for 4 weeks during the run-in period. On day 1 participants will be instructed to take 2 capsules of B. breve strain in the morning before breakfast for 12 weeks. B. breve: Probiotic capsule. Participants will be instructed to take 2 capsules of placebo for 4 weeks during the run-in period. On day 1 participants will be instructed to take 2 capsules of B. breve strain in the morning before breakfast for 12 weeks.	Placebo n=22 Participants Placebo delivered in capsule format. Participants will be instructed to take 2 capsules of placebo for 4 weeks during the run-in period. On day 1 participants will be instructed to take 2 capsules of placebo in the morning before breakfast for 12 weeks. Placebo: Placebo capsule. Participants will be instructed to take 2 capsules of placebo for 4 weeks during the run-in period. On day 1 participants will be instructed to take 2 capsules of placebo in the morning before breakfast for 12 weeks.	Enterotype 2-B. Breve n=10 Participants All subjects (regardless of B. breve or Placebo administration) are divided by enterotype. This sub-arm includes the subjects who share the same Enterotype 2, clustered at the genus level.	Enterotype 2-Placebo n=15 Participants All subjects (regardless of B. breve or Placebo administration) are divided by enterotype. This sub-arm includes the subjects who share the same Enterotype 2, clustered at the genus level.	Enterotype 3-B. Breve n=7 Participants All subjects (regardless of B. breve or Placebo administration) are divided by enterotype. This sub-arm includes the subjects who share the same Enterotype 3, clustered at the genus level.	Enterotype 3-Placebo n=3 Participants All subjects (regardless of B. breve or Placebo administration) are divided by enterotype. This sub-arm includes the subjects who share the same Enterotype 3, clustered at the genus level.
Change in Muscle Mass (g) in Participant Groups Classified by Microbiota Composition at Week 0	108.28 g Standard Deviation 1273.45	364.82 g Standard Deviation 981.21	600.4 g Standard Deviation 1660.45	-82.87 g Standard Deviation 1441.46	-580.57 g Standard Deviation 1202.31	550 g Standard Deviation 641.47

SECONDARY outcome

Timeframe: 12 weeks from baseline

Population: Per-protocol population after excluding the participants with concomitant medication or foods according to the judgement of principal investigator.

The change from baseline between B. breve and placebo in muscle mass (%) in participant groups classified by microbiota composition at week 0, after 12 weeks of supplementation. These individual outcomes comprise relevant aspects of body composition. Each outcome will be analyzed separately and interpreted separately as well as collectively to inform relevant changes to body composition as a whole during the study.

Outcome measures

Measure	B. Breve n=25 Participants Capsule containing B breve strain. Participants will be instructed to take 2 capsules of placebo for 4 weeks during the run-in period. On day 1 participants will be instructed to take 2 capsules of B. breve strain in the morning before breakfast for 12 weeks. B. breve: Probiotic capsule. Participants will be instructed to take 2 capsules of placebo for 4 weeks during the run-in period. On day 1 participants will be instructed to take 2 capsules of B. breve strain in the morning before breakfast for 12 weeks.	Placebo n=22 Participants Placebo delivered in capsule format. Participants will be instructed to take 2 capsules of placebo for 4 weeks during the run-in period. On day 1 participants will be instructed to take 2 capsules of placebo in the morning before breakfast for 12 weeks. Placebo: Placebo capsule. Participants will be instructed to take 2 capsules of placebo for 4 weeks during the run-in period. On day 1 participants will be instructed to take 2 capsules of placebo in the morning before breakfast for 12 weeks.	Enterotype 2-B. Breve n=10 Participants All subjects (regardless of B. breve or Placebo administration) are divided by enterotype. This sub-arm includes the subjects who share the same Enterotype 2, clustered at the genus level.	Enterotype 2-Placebo n=15 Participants All subjects (regardless of B. breve or Placebo administration) are divided by enterotype. This sub-arm includes the subjects who share the same Enterotype 2, clustered at the genus level.	Enterotype 3-B. Breve n=7 Participants All subjects (regardless of B. breve or Placebo administration) are divided by enterotype. This sub-arm includes the subjects who share the same Enterotype 3, clustered at the genus level.	Enterotype 3-Placebo n=3 Participants All subjects (regardless of B. breve or Placebo administration) are divided by enterotype. This sub-arm includes the subjects who share the same Enterotype 3, clustered at the genus level.
Change in Muscle Mass (% of Body Weight) in Participant Groups Classified by Microbiota Composition at Week 0	0.31 percentage of body weight Standard Deviation 2.04	0.2 percentage of body weight Standard Deviation 1.04	0.26 percentage of body weight Standard Deviation 2.28	-0.12 percentage of body weight Standard Deviation 1.7	-0.48 percentage of body weight Standard Deviation 1.86	1.18 percentage of body weight Standard Deviation 1.84

SECONDARY outcome

Timeframe: 12 weeks from baseline

Population: Per-protocol population after excluding the participants with concomitant medication or foods according to the judgement of principal investigator.

The change from baseline between B. breve and placebo in android fat (g) in participant groups classified by microbiota composition at week 0, after 12 weeks of supplementation. These individual outcomes comprise relevant aspects of body composition. Each outcome will be analyzed separately and interpreted separately as well as collectively to inform relevant changes to body composition as a whole during the study.

Outcome measures

Measure	B. Breve n=25 Participants Capsule containing B breve strain. Participants will be instructed to take 2 capsules of placebo for 4 weeks during the run-in period. On day 1 participants will be instructed to take 2 capsules of B. breve strain in the morning before breakfast for 12 weeks. B. breve: Probiotic capsule. Participants will be instructed to take 2 capsules of placebo for 4 weeks during the run-in period. On day 1 participants will be instructed to take 2 capsules of B. breve strain in the morning before breakfast for 12 weeks.	Placebo n=22 Participants Placebo delivered in capsule format. Participants will be instructed to take 2 capsules of placebo for 4 weeks during the run-in period. On day 1 participants will be instructed to take 2 capsules of placebo in the morning before breakfast for 12 weeks. Placebo: Placebo capsule. Participants will be instructed to take 2 capsules of placebo for 4 weeks during the run-in period. On day 1 participants will be instructed to take 2 capsules of placebo in the morning before breakfast for 12 weeks.	Enterotype 2-B. Breve n=10 Participants All subjects (regardless of B. breve or Placebo administration) are divided by enterotype. This sub-arm includes the subjects who share the same Enterotype 2, clustered at the genus level.	Enterotype 2-Placebo n=15 Participants All subjects (regardless of B. breve or Placebo administration) are divided by enterotype. This sub-arm includes the subjects who share the same Enterotype 2, clustered at the genus level.	Enterotype 3-B. Breve n=7 Participants All subjects (regardless of B. breve or Placebo administration) are divided by enterotype. This sub-arm includes the subjects who share the same Enterotype 3, clustered at the genus level.	Enterotype 3-Placebo n=3 Participants All subjects (regardless of B. breve or Placebo administration) are divided by enterotype. This sub-arm includes the subjects who share the same Enterotype 3, clustered at the genus level.
Change in Android Fat (g) in Participant Groups Classified by Microbiota Composition at Week 0	-0.8 g Standard Deviation 269.1	83.14 g Standard Deviation 217.59	-37.1 g Standard Deviation 411.6	1.13 g Standard Deviation 119.24	-24.43 g Standard Deviation 190.81	-122 g Standard Deviation 65.64

SECONDARY outcome

Timeframe: 12 weeks from baseline

Population: Per-protocol population after excluding the participants with concomitant medication or foods according to the judgement of principal investigator.

The change from baseline between B. breve and placebo in android fat (%) in participant groups classified by microbiota composition at week 0, after 12 weeks of supplementation. These individual outcomes comprise relevant aspects of body composition. Each outcome will be analyzed separately and interpreted separately as well as collectively to inform relevant changes to body composition as a whole during the study.

Outcome measures

Measure	B. Breve n=25 Participants Capsule containing B breve strain. Participants will be instructed to take 2 capsules of placebo for 4 weeks during the run-in period. On day 1 participants will be instructed to take 2 capsules of B. breve strain in the morning before breakfast for 12 weeks. B. breve: Probiotic capsule. Participants will be instructed to take 2 capsules of placebo for 4 weeks during the run-in period. On day 1 participants will be instructed to take 2 capsules of B. breve strain in the morning before breakfast for 12 weeks.	Placebo n=22 Participants Placebo delivered in capsule format. Participants will be instructed to take 2 capsules of placebo for 4 weeks during the run-in period. On day 1 participants will be instructed to take 2 capsules of placebo in the morning before breakfast for 12 weeks. Placebo: Placebo capsule. Participants will be instructed to take 2 capsules of placebo for 4 weeks during the run-in period. On day 1 participants will be instructed to take 2 capsules of placebo in the morning before breakfast for 12 weeks.	Enterotype 2-B. Breve n=10 Participants All subjects (regardless of B. breve or Placebo administration) are divided by enterotype. This sub-arm includes the subjects who share the same Enterotype 2, clustered at the genus level.	Enterotype 2-Placebo n=15 Participants All subjects (regardless of B. breve or Placebo administration) are divided by enterotype. This sub-arm includes the subjects who share the same Enterotype 2, clustered at the genus level.	Enterotype 3-B. Breve n=7 Participants All subjects (regardless of B. breve or Placebo administration) are divided by enterotype. This sub-arm includes the subjects who share the same Enterotype 3, clustered at the genus level.	Enterotype 3-Placebo n=3 Participants All subjects (regardless of B. breve or Placebo administration) are divided by enterotype. This sub-arm includes the subjects who share the same Enterotype 3, clustered at the genus level.
Change in Android Fat (% of Android Tissue) in Participant Groups Classified by Microbiota Composition at Week 0	-0.56 percentage of android tissue Standard Deviation 2.8	0.53 percentage of android tissue Standard Deviation 1.17	-0.84 percentage of android tissue Standard Deviation 4.48	0.26 percentage of android tissue Standard Deviation 1.92	0.33 percentage of android tissue Standard Deviation 2.7	-1.7 percentage of android tissue Standard Deviation 0.87

SECONDARY outcome

Timeframe: 12 weeks from baseline

Population: Per-protocol population after excluding the participants with concomitant medication or foods according to the judgement of principal investigator.

The change from baseline between B. breve and placebo in gynoid fat (g) in participant groups classified by microbiota composition at week 0, after 12 weeks of supplementation. These individual outcomes comprise relevant aspects of body composition. Each outcome will be analyzed separately and interpreted separately as well as collectively to inform relevant changes to body composition as a whole during the study.

Outcome measures

Measure	B. Breve n=25 Participants Capsule containing B breve strain. Participants will be instructed to take 2 capsules of placebo for 4 weeks during the run-in period. On day 1 participants will be instructed to take 2 capsules of B. breve strain in the morning before breakfast for 12 weeks. B. breve: Probiotic capsule. Participants will be instructed to take 2 capsules of placebo for 4 weeks during the run-in period. On day 1 participants will be instructed to take 2 capsules of B. breve strain in the morning before breakfast for 12 weeks.	Placebo n=22 Participants Placebo delivered in capsule format. Participants will be instructed to take 2 capsules of placebo for 4 weeks during the run-in period. On day 1 participants will be instructed to take 2 capsules of placebo in the morning before breakfast for 12 weeks. Placebo: Placebo capsule. Participants will be instructed to take 2 capsules of placebo for 4 weeks during the run-in period. On day 1 participants will be instructed to take 2 capsules of placebo in the morning before breakfast for 12 weeks.	Enterotype 2-B. Breve n=10 Participants All subjects (regardless of B. breve or Placebo administration) are divided by enterotype. This sub-arm includes the subjects who share the same Enterotype 2, clustered at the genus level.	Enterotype 2-Placebo n=15 Participants All subjects (regardless of B. breve or Placebo administration) are divided by enterotype. This sub-arm includes the subjects who share the same Enterotype 2, clustered at the genus level.	Enterotype 3-B. Breve n=7 Participants All subjects (regardless of B. breve or Placebo administration) are divided by enterotype. This sub-arm includes the subjects who share the same Enterotype 3, clustered at the genus level.	Enterotype 3-Placebo n=3 Participants All subjects (regardless of B. breve or Placebo administration) are divided by enterotype. This sub-arm includes the subjects who share the same Enterotype 3, clustered at the genus level.
Change in Gynoid Fat (g) in Participant Groups Classified by Microbiota Composition at Week 0	83.52 g Standard Deviation 555.31	75.09 g Standard Deviation 310.83	94.50 g Standard Deviation 452.95	-4.13 g Standard Deviation 524.2	107.29 g Standard Deviation 420.15	36 g Standard Deviation 677.02

SECONDARY outcome

Timeframe: 12 weeks from baseline

Population: Per-protocol population after excluding the participants with concomitant medication or foods according to the judgement of principal investigator.

The change from baseline between B. breve and placebo in gynoid fat (%) in participant groups classified by microbiota composition at week 0, after 12 weeks of supplementation. These individual outcomes comprise relevant aspects of body composition. Each outcome will be analyzed separately and interpreted separately as well as collectively to inform relevant changes to body composition as a whole during the study.

Outcome measures

Measure	B. Breve n=25 Participants Capsule containing B breve strain. Participants will be instructed to take 2 capsules of placebo for 4 weeks during the run-in period. On day 1 participants will be instructed to take 2 capsules of B. breve strain in the morning before breakfast for 12 weeks. B. breve: Probiotic capsule. Participants will be instructed to take 2 capsules of placebo for 4 weeks during the run-in period. On day 1 participants will be instructed to take 2 capsules of B. breve strain in the morning before breakfast for 12 weeks.	Placebo n=22 Participants Placebo delivered in capsule format. Participants will be instructed to take 2 capsules of placebo for 4 weeks during the run-in period. On day 1 participants will be instructed to take 2 capsules of placebo in the morning before breakfast for 12 weeks. Placebo: Placebo capsule. Participants will be instructed to take 2 capsules of placebo for 4 weeks during the run-in period. On day 1 participants will be instructed to take 2 capsules of placebo in the morning before breakfast for 12 weeks.	Enterotype 2-B. Breve n=10 Participants All subjects (regardless of B. breve or Placebo administration) are divided by enterotype. This sub-arm includes the subjects who share the same Enterotype 2, clustered at the genus level.	Enterotype 2-Placebo n=15 Participants All subjects (regardless of B. breve or Placebo administration) are divided by enterotype. This sub-arm includes the subjects who share the same Enterotype 2, clustered at the genus level.	Enterotype 3-B. Breve n=7 Participants All subjects (regardless of B. breve or Placebo administration) are divided by enterotype. This sub-arm includes the subjects who share the same Enterotype 3, clustered at the genus level.	Enterotype 3-Placebo n=3 Participants All subjects (regardless of B. breve or Placebo administration) are divided by enterotype. This sub-arm includes the subjects who share the same Enterotype 3, clustered at the genus level.
Change in Gynoid Fat (% of Gynoid Tissue) in Participant Groups Classified by Microbiota Composition at Week 0	0.2 percentage of gynoid tissue Standard Deviation 2.48	0.28 percentage of gynoid tissue Standard Deviation 1.75	0.74 percentage of gynoid tissue Standard Deviation 2.44	0.1 percentage of gynoid tissue Standard Deviation 2.65	0.36 percentage of gynoid tissue Standard Deviation 2.51	-0.57 percentage of gynoid tissue Standard Deviation 1.6

SECONDARY outcome

Timeframe: 12 weeks from baseline

Population: Per-protocol population after excluding the participants with concomitant medication or foods according to the judgement of principal investigator.

The change from baseline between B. breve and placebo in android:gynoid fat ratio in participant groups classified by microbiota composition at week 0, after 12 weeks of supplementation. These individual outcomes comprise relevant aspects of body composition. Each outcome will be analyzed separately and interpreted separately as well as collectively to inform relevant changes to body composition as a whole during the study.

Outcome measures

Measure	B. Breve n=25 Participants Capsule containing B breve strain. Participants will be instructed to take 2 capsules of placebo for 4 weeks during the run-in period. On day 1 participants will be instructed to take 2 capsules of B. breve strain in the morning before breakfast for 12 weeks. B. breve: Probiotic capsule. Participants will be instructed to take 2 capsules of placebo for 4 weeks during the run-in period. On day 1 participants will be instructed to take 2 capsules of B. breve strain in the morning before breakfast for 12 weeks.	Placebo n=22 Participants Placebo delivered in capsule format. Participants will be instructed to take 2 capsules of placebo for 4 weeks during the run-in period. On day 1 participants will be instructed to take 2 capsules of placebo in the morning before breakfast for 12 weeks. Placebo: Placebo capsule. Participants will be instructed to take 2 capsules of placebo for 4 weeks during the run-in period. On day 1 participants will be instructed to take 2 capsules of placebo in the morning before breakfast for 12 weeks.	Enterotype 2-B. Breve n=10 Participants All subjects (regardless of B. breve or Placebo administration) are divided by enterotype. This sub-arm includes the subjects who share the same Enterotype 2, clustered at the genus level.	Enterotype 2-Placebo n=15 Participants All subjects (regardless of B. breve or Placebo administration) are divided by enterotype. This sub-arm includes the subjects who share the same Enterotype 2, clustered at the genus level.	Enterotype 3-B. Breve n=7 Participants All subjects (regardless of B. breve or Placebo administration) are divided by enterotype. This sub-arm includes the subjects who share the same Enterotype 3, clustered at the genus level.	Enterotype 3-Placebo n=3 Participants All subjects (regardless of B. breve or Placebo administration) are divided by enterotype. This sub-arm includes the subjects who share the same Enterotype 3, clustered at the genus level.
Change in Android:Gynoid Fat Ratio in Participant Groups Classified by Microbiota Composition at Week 0	-0.01 percentage of gynoid fat Standard Deviation 0.05	0.01 percentage of gynoid fat Standard Deviation 0.06	-0.04 percentage of gynoid fat Standard Deviation 0.1	0.00 percentage of gynoid fat Standard Deviation 0.06	0.02 percentage of gynoid fat Standard Deviation 0.04	-0.03 percentage of gynoid fat Standard Deviation 0.02

SECONDARY outcome

Timeframe: 12 weeks from baseline

Population: Per-protocol population after excluding the participants with concomitant medication or foods according to the judgement of principal investigator.

The change from baseline between B. breve and placebo in waist circumference in participant groups classified by microbiota composition at week 0, after 12 weeks of supplementation. These individual outcomes comprise relevant aspects of body composition. Each outcome will be analyzed separately and interpreted separately as well as collectively to inform relevant changes to body composition as a whole during the study.

Outcome measures

Measure	B. Breve n=25 Participants Capsule containing B breve strain. Participants will be instructed to take 2 capsules of placebo for 4 weeks during the run-in period. On day 1 participants will be instructed to take 2 capsules of B. breve strain in the morning before breakfast for 12 weeks. B. breve: Probiotic capsule. Participants will be instructed to take 2 capsules of placebo for 4 weeks during the run-in period. On day 1 participants will be instructed to take 2 capsules of B. breve strain in the morning before breakfast for 12 weeks.	Placebo n=22 Participants Placebo delivered in capsule format. Participants will be instructed to take 2 capsules of placebo for 4 weeks during the run-in period. On day 1 participants will be instructed to take 2 capsules of placebo in the morning before breakfast for 12 weeks. Placebo: Placebo capsule. Participants will be instructed to take 2 capsules of placebo for 4 weeks during the run-in period. On day 1 participants will be instructed to take 2 capsules of placebo in the morning before breakfast for 12 weeks.	Enterotype 2-B. Breve n=10 Participants All subjects (regardless of B. breve or Placebo administration) are divided by enterotype. This sub-arm includes the subjects who share the same Enterotype 2, clustered at the genus level.	Enterotype 2-Placebo n=15 Participants All subjects (regardless of B. breve or Placebo administration) are divided by enterotype. This sub-arm includes the subjects who share the same Enterotype 2, clustered at the genus level.	Enterotype 3-B. Breve n=7 Participants All subjects (regardless of B. breve or Placebo administration) are divided by enterotype. This sub-arm includes the subjects who share the same Enterotype 3, clustered at the genus level.	Enterotype 3-Placebo n=3 Participants All subjects (regardless of B. breve or Placebo administration) are divided by enterotype. This sub-arm includes the subjects who share the same Enterotype 3, clustered at the genus level.
Change in Waist Circumference in Participant Groups Classified by Microbiota Composition at Week 0	-0.36 cm Standard Deviation 3.46	0.00 cm Standard Deviation 3.41	0.63 cm Standard Deviation 2.38	-0.94 cm Standard Deviation 3.29	0.03 cm Standard Deviation 0.93	-2.4 cm Standard Deviation 3.4

SECONDARY outcome

Timeframe: 12 weeks from baseline

Population: Per-protocol population after excluding the participants with concomitant medication or foods according to the judgement of principal investigator.

The change from baseline between B. breve and placebo in hip circumference, in participant groups classified by microbiota composition at week 0, after 12 weeks of supplementation. These individual outcomes comprise relevant aspects of body composition. Each outcome will be analyzed separately and interpreted separately as well as collectively to inform relevant changes to body composition as a whole during the study.

Outcome measures

Measure	B. Breve n=25 Participants Capsule containing B breve strain. Participants will be instructed to take 2 capsules of placebo for 4 weeks during the run-in period. On day 1 participants will be instructed to take 2 capsules of B. breve strain in the morning before breakfast for 12 weeks. B. breve: Probiotic capsule. Participants will be instructed to take 2 capsules of placebo for 4 weeks during the run-in period. On day 1 participants will be instructed to take 2 capsules of B. breve strain in the morning before breakfast for 12 weeks.	Placebo n=22 Participants Placebo delivered in capsule format. Participants will be instructed to take 2 capsules of placebo for 4 weeks during the run-in period. On day 1 participants will be instructed to take 2 capsules of placebo in the morning before breakfast for 12 weeks. Placebo: Placebo capsule. Participants will be instructed to take 2 capsules of placebo for 4 weeks during the run-in period. On day 1 participants will be instructed to take 2 capsules of placebo in the morning before breakfast for 12 weeks.	Enterotype 2-B. Breve n=10 Participants All subjects (regardless of B. breve or Placebo administration) are divided by enterotype. This sub-arm includes the subjects who share the same Enterotype 2, clustered at the genus level.	Enterotype 2-Placebo n=15 Participants All subjects (regardless of B. breve or Placebo administration) are divided by enterotype. This sub-arm includes the subjects who share the same Enterotype 2, clustered at the genus level.	Enterotype 3-B. Breve n=7 Participants All subjects (regardless of B. breve or Placebo administration) are divided by enterotype. This sub-arm includes the subjects who share the same Enterotype 3, clustered at the genus level.	Enterotype 3-Placebo n=3 Participants All subjects (regardless of B. breve or Placebo administration) are divided by enterotype. This sub-arm includes the subjects who share the same Enterotype 3, clustered at the genus level.
Change in Hip Circumference in Participant Groups Classified by Microbiota Composition at Week 0	-1.08 cm Standard Deviation 4.08	-0.26 cm Standard Deviation 5.08	-1.07 cm Standard Deviation 3.3	-1.77 cm Standard Deviation 4.04	-0.86 cm Standard Deviation 2.13	-2.1 cm Standard Deviation 0.17

SECONDARY outcome

Timeframe: 12 weeks from baseline

Population: Per-protocol population after excluding the participants with concomitant medication or foods according to the judgement of principal investigator.

The change from baseline between B. breve and placebo in waist:hip circumference ratio in participant groups classified by microbiota composition at week 0, after 12 weeks of supplementation. These individual outcomes comprise relevant aspects of body composition. Each outcome will be analyzed separately and interpreted separately as well as collectively to inform relevant changes to body composition as a whole during the study.

Outcome measures

Measure	B. Breve n=25 Participants Capsule containing B breve strain. Participants will be instructed to take 2 capsules of placebo for 4 weeks during the run-in period. On day 1 participants will be instructed to take 2 capsules of B. breve strain in the morning before breakfast for 12 weeks. B. breve: Probiotic capsule. Participants will be instructed to take 2 capsules of placebo for 4 weeks during the run-in period. On day 1 participants will be instructed to take 2 capsules of B. breve strain in the morning before breakfast for 12 weeks.	Placebo n=22 Participants Placebo delivered in capsule format. Participants will be instructed to take 2 capsules of placebo for 4 weeks during the run-in period. On day 1 participants will be instructed to take 2 capsules of placebo in the morning before breakfast for 12 weeks. Placebo: Placebo capsule. Participants will be instructed to take 2 capsules of placebo for 4 weeks during the run-in period. On day 1 participants will be instructed to take 2 capsules of placebo in the morning before breakfast for 12 weeks.	Enterotype 2-B. Breve n=10 Participants All subjects (regardless of B. breve or Placebo administration) are divided by enterotype. This sub-arm includes the subjects who share the same Enterotype 2, clustered at the genus level.	Enterotype 2-Placebo n=15 Participants All subjects (regardless of B. breve or Placebo administration) are divided by enterotype. This sub-arm includes the subjects who share the same Enterotype 2, clustered at the genus level.	Enterotype 3-B. Breve n=7 Participants All subjects (regardless of B. breve or Placebo administration) are divided by enterotype. This sub-arm includes the subjects who share the same Enterotype 3, clustered at the genus level.	Enterotype 3-Placebo n=3 Participants All subjects (regardless of B. breve or Placebo administration) are divided by enterotype. This sub-arm includes the subjects who share the same Enterotype 3, clustered at the genus level.
Change in Waist:Hip Circumference Ratio in Participant Groups Classified by Microbiota Composition at Week 0	0.01 percentage of hip circumference Standard Deviation 0.05	0.00 percentage of hip circumference Standard Deviation 0.06	0.02 percentage of hip circumference Standard Deviation 0.04	0.01 percentage of hip circumference Standard Deviation 0.03	0.01 percentage of hip circumference Standard Deviation 0.01	-0.01 percentage of hip circumference Standard Deviation 0.04

SECONDARY outcome

Timeframe: Baseline and 12 weeks of supplementation

Population: Per-protocol population after excluding the participants with concomitant medication or foods according to the judgement of principal investigator.

The difference in change from baseline between B. breve and placebo in total cholesterol after 12 weeks of supplementation.

Outcome measures

Measure	B. Breve n=42 Participants Capsule containing B breve strain. Participants will be instructed to take 2 capsules of placebo for 4 weeks during the run-in period. On day 1 participants will be instructed to take 2 capsules of B. breve strain in the morning before breakfast for 12 weeks. B. breve: Probiotic capsule. Participants will be instructed to take 2 capsules of placebo for 4 weeks during the run-in period. On day 1 participants will be instructed to take 2 capsules of B. breve strain in the morning before breakfast for 12 weeks.	Placebo n=40 Participants Placebo delivered in capsule format. Participants will be instructed to take 2 capsules of placebo for 4 weeks during the run-in period. On day 1 participants will be instructed to take 2 capsules of placebo in the morning before breakfast for 12 weeks. Placebo: Placebo capsule. Participants will be instructed to take 2 capsules of placebo for 4 weeks during the run-in period. On day 1 participants will be instructed to take 2 capsules of placebo in the morning before breakfast for 12 weeks.	Enterotype 2-B. Breve All subjects (regardless of B. breve or Placebo administration) are divided by enterotype. This sub-arm includes the subjects who share the same Enterotype 2, clustered at the genus level.	Enterotype 2-Placebo All subjects (regardless of B. breve or Placebo administration) are divided by enterotype. This sub-arm includes the subjects who share the same Enterotype 2, clustered at the genus level.	Enterotype 3-B. Breve All subjects (regardless of B. breve or Placebo administration) are divided by enterotype. This sub-arm includes the subjects who share the same Enterotype 3, clustered at the genus level.	Enterotype 3-Placebo All subjects (regardless of B. breve or Placebo administration) are divided by enterotype. This sub-arm includes the subjects who share the same Enterotype 3, clustered at the genus level.
Total Cholesterol	-0.05 mmol/L Standard Deviation 0.69	0.08 mmol/L Standard Deviation 0.53	—	—	—	—

SECONDARY outcome

Timeframe: Baseline and 12 weeks of supplementation

Population: Per-protocol population after excluding the participants with concomitant medication or foods according to the judgement of principal investigator.

The difference in change from baseline between B. breve and placebo in HDL-cholesterol after 12 weeks of supplementation.

Outcome measures

Measure	B. Breve n=42 Participants Capsule containing B breve strain. Participants will be instructed to take 2 capsules of placebo for 4 weeks during the run-in period. On day 1 participants will be instructed to take 2 capsules of B. breve strain in the morning before breakfast for 12 weeks. B. breve: Probiotic capsule. Participants will be instructed to take 2 capsules of placebo for 4 weeks during the run-in period. On day 1 participants will be instructed to take 2 capsules of B. breve strain in the morning before breakfast for 12 weeks.	Placebo n=40 Participants Placebo delivered in capsule format. Participants will be instructed to take 2 capsules of placebo for 4 weeks during the run-in period. On day 1 participants will be instructed to take 2 capsules of placebo in the morning before breakfast for 12 weeks. Placebo: Placebo capsule. Participants will be instructed to take 2 capsules of placebo for 4 weeks during the run-in period. On day 1 participants will be instructed to take 2 capsules of placebo in the morning before breakfast for 12 weeks.	Enterotype 2-B. Breve All subjects (regardless of B. breve or Placebo administration) are divided by enterotype. This sub-arm includes the subjects who share the same Enterotype 2, clustered at the genus level.	Enterotype 2-Placebo All subjects (regardless of B. breve or Placebo administration) are divided by enterotype. This sub-arm includes the subjects who share the same Enterotype 2, clustered at the genus level.	Enterotype 3-B. Breve All subjects (regardless of B. breve or Placebo administration) are divided by enterotype. This sub-arm includes the subjects who share the same Enterotype 3, clustered at the genus level.	Enterotype 3-Placebo All subjects (regardless of B. breve or Placebo administration) are divided by enterotype. This sub-arm includes the subjects who share the same Enterotype 3, clustered at the genus level.
HDL-cholesterol	-0.03 mmol/L Standard Deviation 0.18	0.00 mmol/L Standard Deviation 0.19	—	—	—	—

SECONDARY outcome

Timeframe: Baseline and 12 weeks of supplementation

Population: Per-protocol population after excluding the participants with concomitant medication or foods according to the judgement of principal investigator.

The difference in change from baseline between B. breve and placebo in LDL-cholesterol after 12 weeks of supplementation.

Outcome measures

Measure	B. Breve n=42 Participants Capsule containing B breve strain. Participants will be instructed to take 2 capsules of placebo for 4 weeks during the run-in period. On day 1 participants will be instructed to take 2 capsules of B. breve strain in the morning before breakfast for 12 weeks. B. breve: Probiotic capsule. Participants will be instructed to take 2 capsules of placebo for 4 weeks during the run-in period. On day 1 participants will be instructed to take 2 capsules of B. breve strain in the morning before breakfast for 12 weeks.	Placebo n=40 Participants Placebo delivered in capsule format. Participants will be instructed to take 2 capsules of placebo for 4 weeks during the run-in period. On day 1 participants will be instructed to take 2 capsules of placebo in the morning before breakfast for 12 weeks. Placebo: Placebo capsule. Participants will be instructed to take 2 capsules of placebo for 4 weeks during the run-in period. On day 1 participants will be instructed to take 2 capsules of placebo in the morning before breakfast for 12 weeks.	Enterotype 2-B. Breve All subjects (regardless of B. breve or Placebo administration) are divided by enterotype. This sub-arm includes the subjects who share the same Enterotype 2, clustered at the genus level.	Enterotype 2-Placebo All subjects (regardless of B. breve or Placebo administration) are divided by enterotype. This sub-arm includes the subjects who share the same Enterotype 2, clustered at the genus level.	Enterotype 3-B. Breve All subjects (regardless of B. breve or Placebo administration) are divided by enterotype. This sub-arm includes the subjects who share the same Enterotype 3, clustered at the genus level.	Enterotype 3-Placebo All subjects (regardless of B. breve or Placebo administration) are divided by enterotype. This sub-arm includes the subjects who share the same Enterotype 3, clustered at the genus level.
LDL-cholesterol	-0.06 mmol/L Standard Deviation 0.57	0.09 mmol/L Standard Deviation 0.47	—	—	—	—

SECONDARY outcome

Timeframe: Baseline and 12 weeks of supplementation

Population: Per-protocol population after excluding the participants with concomitant medication or foods according to the judgement of principal investigator.

The difference in change from baseline between B. breve and placebo in triglycerides after 12 weeks of supplementation.

Outcome measures

Measure	B. Breve n=42 Participants Capsule containing B breve strain. Participants will be instructed to take 2 capsules of placebo for 4 weeks during the run-in period. On day 1 participants will be instructed to take 2 capsules of B. breve strain in the morning before breakfast for 12 weeks. B. breve: Probiotic capsule. Participants will be instructed to take 2 capsules of placebo for 4 weeks during the run-in period. On day 1 participants will be instructed to take 2 capsules of B. breve strain in the morning before breakfast for 12 weeks.	Placebo n=40 Participants Placebo delivered in capsule format. Participants will be instructed to take 2 capsules of placebo for 4 weeks during the run-in period. On day 1 participants will be instructed to take 2 capsules of placebo in the morning before breakfast for 12 weeks. Placebo: Placebo capsule. Participants will be instructed to take 2 capsules of placebo for 4 weeks during the run-in period. On day 1 participants will be instructed to take 2 capsules of placebo in the morning before breakfast for 12 weeks.	Enterotype 2-B. Breve All subjects (regardless of B. breve or Placebo administration) are divided by enterotype. This sub-arm includes the subjects who share the same Enterotype 2, clustered at the genus level.	Enterotype 2-Placebo All subjects (regardless of B. breve or Placebo administration) are divided by enterotype. This sub-arm includes the subjects who share the same Enterotype 2, clustered at the genus level.	Enterotype 3-B. Breve All subjects (regardless of B. breve or Placebo administration) are divided by enterotype. This sub-arm includes the subjects who share the same Enterotype 3, clustered at the genus level.	Enterotype 3-Placebo All subjects (regardless of B. breve or Placebo administration) are divided by enterotype. This sub-arm includes the subjects who share the same Enterotype 3, clustered at the genus level.
Triglycerides	0.09 mmol/L Standard Deviation 0.54	-0.04 mmol/L Standard Deviation 0.50	—	—	—	—

SECONDARY outcome

Timeframe: Baseline and 12 weeks of supplementation

Population: Per-protocol population after excluding the participants with concomitant medication or foods according to the judgement of principal investigator.

The difference in change from baseline between B. breve and placebo in fasting blood glucose after 12 weeks of supplementation.

Outcome measures

Measure	B. Breve n=42 Participants Capsule containing B breve strain. Participants will be instructed to take 2 capsules of placebo for 4 weeks during the run-in period. On day 1 participants will be instructed to take 2 capsules of B. breve strain in the morning before breakfast for 12 weeks. B. breve: Probiotic capsule. Participants will be instructed to take 2 capsules of placebo for 4 weeks during the run-in period. On day 1 participants will be instructed to take 2 capsules of B. breve strain in the morning before breakfast for 12 weeks.	Placebo n=40 Participants Placebo delivered in capsule format. Participants will be instructed to take 2 capsules of placebo for 4 weeks during the run-in period. On day 1 participants will be instructed to take 2 capsules of placebo in the morning before breakfast for 12 weeks. Placebo: Placebo capsule. Participants will be instructed to take 2 capsules of placebo for 4 weeks during the run-in period. On day 1 participants will be instructed to take 2 capsules of placebo in the morning before breakfast for 12 weeks.	Enterotype 2-B. Breve All subjects (regardless of B. breve or Placebo administration) are divided by enterotype. This sub-arm includes the subjects who share the same Enterotype 2, clustered at the genus level.	Enterotype 2-Placebo All subjects (regardless of B. breve or Placebo administration) are divided by enterotype. This sub-arm includes the subjects who share the same Enterotype 2, clustered at the genus level.	Enterotype 3-B. Breve All subjects (regardless of B. breve or Placebo administration) are divided by enterotype. This sub-arm includes the subjects who share the same Enterotype 3, clustered at the genus level.	Enterotype 3-Placebo All subjects (regardless of B. breve or Placebo administration) are divided by enterotype. This sub-arm includes the subjects who share the same Enterotype 3, clustered at the genus level.
Fasting Blood Glucose	0.05 mmol/L Standard Deviation 0.39	0.15 mmol/L Standard Deviation 0.33	—	—	—	—

SECONDARY outcome

Timeframe: Baseline and 12 weeks of supplementation

Population: Per-protocol population after excluding the participants with concomitant medication or foods according to the judgement of principal investigator.

The difference in change from baseline between B. breve and placebo in HbA1c after 12 weeks of supplementation.

Outcome measures

Measure	B. Breve n=42 Participants Capsule containing B breve strain. Participants will be instructed to take 2 capsules of placebo for 4 weeks during the run-in period. On day 1 participants will be instructed to take 2 capsules of B. breve strain in the morning before breakfast for 12 weeks. B. breve: Probiotic capsule. Participants will be instructed to take 2 capsules of placebo for 4 weeks during the run-in period. On day 1 participants will be instructed to take 2 capsules of B. breve strain in the morning before breakfast for 12 weeks.	Placebo n=40 Participants Placebo delivered in capsule format. Participants will be instructed to take 2 capsules of placebo for 4 weeks during the run-in period. On day 1 participants will be instructed to take 2 capsules of placebo in the morning before breakfast for 12 weeks. Placebo: Placebo capsule. Participants will be instructed to take 2 capsules of placebo for 4 weeks during the run-in period. On day 1 participants will be instructed to take 2 capsules of placebo in the morning before breakfast for 12 weeks.	Enterotype 2-B. Breve All subjects (regardless of B. breve or Placebo administration) are divided by enterotype. This sub-arm includes the subjects who share the same Enterotype 2, clustered at the genus level.	Enterotype 2-Placebo All subjects (regardless of B. breve or Placebo administration) are divided by enterotype. This sub-arm includes the subjects who share the same Enterotype 2, clustered at the genus level.	Enterotype 3-B. Breve All subjects (regardless of B. breve or Placebo administration) are divided by enterotype. This sub-arm includes the subjects who share the same Enterotype 3, clustered at the genus level.	Enterotype 3-Placebo All subjects (regardless of B. breve or Placebo administration) are divided by enterotype. This sub-arm includes the subjects who share the same Enterotype 3, clustered at the genus level.
HbA1c	0.02 percentage of glycated hemoglobin Standard Deviation 0.17	0.03 percentage of glycated hemoglobin Standard Deviation 0.17	—	—	—	—

SECONDARY outcome

Timeframe: Baseline and 12 weeks of supplementation

Population: Per-protocol population after excluding the participants with concomitant medication or foods according to the judgement of principal investigator.

The difference in change from baseline between B. breve and placebo in fasting insulin after 12 weeks of supplementation.

Outcome measures

Measure	B. Breve n=42 Participants Capsule containing B breve strain. Participants will be instructed to take 2 capsules of placebo for 4 weeks during the run-in period. On day 1 participants will be instructed to take 2 capsules of B. breve strain in the morning before breakfast for 12 weeks. B. breve: Probiotic capsule. Participants will be instructed to take 2 capsules of placebo for 4 weeks during the run-in period. On day 1 participants will be instructed to take 2 capsules of B. breve strain in the morning before breakfast for 12 weeks.	Placebo n=40 Participants Placebo delivered in capsule format. Participants will be instructed to take 2 capsules of placebo for 4 weeks during the run-in period. On day 1 participants will be instructed to take 2 capsules of placebo in the morning before breakfast for 12 weeks. Placebo: Placebo capsule. Participants will be instructed to take 2 capsules of placebo for 4 weeks during the run-in period. On day 1 participants will be instructed to take 2 capsules of placebo in the morning before breakfast for 12 weeks.	Enterotype 2-B. Breve All subjects (regardless of B. breve or Placebo administration) are divided by enterotype. This sub-arm includes the subjects who share the same Enterotype 2, clustered at the genus level.	Enterotype 2-Placebo All subjects (regardless of B. breve or Placebo administration) are divided by enterotype. This sub-arm includes the subjects who share the same Enterotype 2, clustered at the genus level.	Enterotype 3-B. Breve All subjects (regardless of B. breve or Placebo administration) are divided by enterotype. This sub-arm includes the subjects who share the same Enterotype 3, clustered at the genus level.	Enterotype 3-Placebo All subjects (regardless of B. breve or Placebo administration) are divided by enterotype. This sub-arm includes the subjects who share the same Enterotype 3, clustered at the genus level.
Fasting Insulin	-9.43 pmol/L Standard Deviation 61.49	4.10 pmol/L Standard Deviation 28.25	—	—	—	—

SECONDARY outcome

Timeframe: Baseline and 12 weeks of supplementation

Population: Per-protocol population after excluding the participants with concomitant medication or foods according to the judgement of principal investigator.

The difference in change from baseline between B. breve and placebo in ALP after 12 weeks of supplementation.

Outcome measures

Measure	B. Breve n=42 Participants Capsule containing B breve strain. Participants will be instructed to take 2 capsules of placebo for 4 weeks during the run-in period. On day 1 participants will be instructed to take 2 capsules of B. breve strain in the morning before breakfast for 12 weeks. B. breve: Probiotic capsule. Participants will be instructed to take 2 capsules of placebo for 4 weeks during the run-in period. On day 1 participants will be instructed to take 2 capsules of B. breve strain in the morning before breakfast for 12 weeks.	Placebo n=40 Participants Placebo delivered in capsule format. Participants will be instructed to take 2 capsules of placebo for 4 weeks during the run-in period. On day 1 participants will be instructed to take 2 capsules of placebo in the morning before breakfast for 12 weeks. Placebo: Placebo capsule. Participants will be instructed to take 2 capsules of placebo for 4 weeks during the run-in period. On day 1 participants will be instructed to take 2 capsules of placebo in the morning before breakfast for 12 weeks.	Enterotype 2-B. Breve All subjects (regardless of B. breve or Placebo administration) are divided by enterotype. This sub-arm includes the subjects who share the same Enterotype 2, clustered at the genus level.	Enterotype 2-Placebo All subjects (regardless of B. breve or Placebo administration) are divided by enterotype. This sub-arm includes the subjects who share the same Enterotype 2, clustered at the genus level.	Enterotype 3-B. Breve All subjects (regardless of B. breve or Placebo administration) are divided by enterotype. This sub-arm includes the subjects who share the same Enterotype 3, clustered at the genus level.	Enterotype 3-Placebo All subjects (regardless of B. breve or Placebo administration) are divided by enterotype. This sub-arm includes the subjects who share the same Enterotype 3, clustered at the genus level.
Alkaline Phosphatase (ALP)	2.83 U/L Standard Deviation 13.21	0.40 U/L Standard Deviation 8.44	—	—	—	—

SECONDARY outcome

Timeframe: Baseline and 12 weeks of supplementation

Population: Per-protocol population after excluding the participants with concomitant medication or foods according to the judgement of principal investigator.

The difference in change from baseline between B. breve and placebo in GGT after 12 weeks of supplementation.

Outcome measures

Measure	B. Breve n=42 Participants Capsule containing B breve strain. Participants will be instructed to take 2 capsules of placebo for 4 weeks during the run-in period. On day 1 participants will be instructed to take 2 capsules of B. breve strain in the morning before breakfast for 12 weeks. B. breve: Probiotic capsule. Participants will be instructed to take 2 capsules of placebo for 4 weeks during the run-in period. On day 1 participants will be instructed to take 2 capsules of B. breve strain in the morning before breakfast for 12 weeks.	Placebo n=40 Participants Placebo delivered in capsule format. Participants will be instructed to take 2 capsules of placebo for 4 weeks during the run-in period. On day 1 participants will be instructed to take 2 capsules of placebo in the morning before breakfast for 12 weeks. Placebo: Placebo capsule. Participants will be instructed to take 2 capsules of placebo for 4 weeks during the run-in period. On day 1 participants will be instructed to take 2 capsules of placebo in the morning before breakfast for 12 weeks.	Enterotype 2-B. Breve All subjects (regardless of B. breve or Placebo administration) are divided by enterotype. This sub-arm includes the subjects who share the same Enterotype 2, clustered at the genus level.	Enterotype 2-Placebo All subjects (regardless of B. breve or Placebo administration) are divided by enterotype. This sub-arm includes the subjects who share the same Enterotype 2, clustered at the genus level.	Enterotype 3-B. Breve All subjects (regardless of B. breve or Placebo administration) are divided by enterotype. This sub-arm includes the subjects who share the same Enterotype 3, clustered at the genus level.	Enterotype 3-Placebo All subjects (regardless of B. breve or Placebo administration) are divided by enterotype. This sub-arm includes the subjects who share the same Enterotype 3, clustered at the genus level.
Gamma-glutamyl Transferase (GGT)	0.81 U/L Standard Deviation 5.58	0.25 U/L Standard Deviation 3.72	—	—	—	—

SECONDARY outcome

Timeframe: Baseline and 12 weeks of supplementation

Population: Per-protocol population after excluding the participants with concomitant medication or foods according to the judgement of principal investigator.

The difference in change from baseline between B. breve and placebo in ALT after 12 weeks of supplementation.

Outcome measures

Measure	B. Breve n=42 Participants Capsule containing B breve strain. Participants will be instructed to take 2 capsules of placebo for 4 weeks during the run-in period. On day 1 participants will be instructed to take 2 capsules of B. breve strain in the morning before breakfast for 12 weeks. B. breve: Probiotic capsule. Participants will be instructed to take 2 capsules of placebo for 4 weeks during the run-in period. On day 1 participants will be instructed to take 2 capsules of B. breve strain in the morning before breakfast for 12 weeks.	Placebo n=40 Participants Placebo delivered in capsule format. Participants will be instructed to take 2 capsules of placebo for 4 weeks during the run-in period. On day 1 participants will be instructed to take 2 capsules of placebo in the morning before breakfast for 12 weeks. Placebo: Placebo capsule. Participants will be instructed to take 2 capsules of placebo for 4 weeks during the run-in period. On day 1 participants will be instructed to take 2 capsules of placebo in the morning before breakfast for 12 weeks.	Enterotype 2-B. Breve All subjects (regardless of B. breve or Placebo administration) are divided by enterotype. This sub-arm includes the subjects who share the same Enterotype 2, clustered at the genus level.	Enterotype 2-Placebo All subjects (regardless of B. breve or Placebo administration) are divided by enterotype. This sub-arm includes the subjects who share the same Enterotype 2, clustered at the genus level.	Enterotype 3-B. Breve All subjects (regardless of B. breve or Placebo administration) are divided by enterotype. This sub-arm includes the subjects who share the same Enterotype 3, clustered at the genus level.	Enterotype 3-Placebo All subjects (regardless of B. breve or Placebo administration) are divided by enterotype. This sub-arm includes the subjects who share the same Enterotype 3, clustered at the genus level.
Alanine Aminotransferase (ALT)	0.60 U/L Standard Deviation 12.72	-1.70 U/L Standard Deviation 7.63	—	—	—	—

SECONDARY outcome

Timeframe: Baseline and 12 weeks of supplementation

Population: Per-protocol population after excluding the participants with concomitant medication or foods according to the judgement of principal investigator.

The difference in change from baseline between B. breve and placebo in AST after 12 weeks of supplementation.

Outcome measures

Measure	B. Breve n=42 Participants Capsule containing B breve strain. Participants will be instructed to take 2 capsules of placebo for 4 weeks during the run-in period. On day 1 participants will be instructed to take 2 capsules of B. breve strain in the morning before breakfast for 12 weeks. B. breve: Probiotic capsule. Participants will be instructed to take 2 capsules of placebo for 4 weeks during the run-in period. On day 1 participants will be instructed to take 2 capsules of B. breve strain in the morning before breakfast for 12 weeks.	Placebo n=40 Participants Placebo delivered in capsule format. Participants will be instructed to take 2 capsules of placebo for 4 weeks during the run-in period. On day 1 participants will be instructed to take 2 capsules of placebo in the morning before breakfast for 12 weeks. Placebo: Placebo capsule. Participants will be instructed to take 2 capsules of placebo for 4 weeks during the run-in period. On day 1 participants will be instructed to take 2 capsules of placebo in the morning before breakfast for 12 weeks.	Enterotype 2-B. Breve All subjects (regardless of B. breve or Placebo administration) are divided by enterotype. This sub-arm includes the subjects who share the same Enterotype 2, clustered at the genus level.	Enterotype 2-Placebo All subjects (regardless of B. breve or Placebo administration) are divided by enterotype. This sub-arm includes the subjects who share the same Enterotype 2, clustered at the genus level.	Enterotype 3-B. Breve All subjects (regardless of B. breve or Placebo administration) are divided by enterotype. This sub-arm includes the subjects who share the same Enterotype 3, clustered at the genus level.	Enterotype 3-Placebo All subjects (regardless of B. breve or Placebo administration) are divided by enterotype. This sub-arm includes the subjects who share the same Enterotype 3, clustered at the genus level.
Aspartate Transaminase (AST)	0.40 U/L Standard Deviation 5.91	-1.77 U/L Standard Deviation 7.66	—	—	—	—

SECONDARY outcome

Timeframe: Baseline, 6 and 12 weeks of supplementation

Population: Per-protocol population after excluding the participants with concomitant medication or foods according to the judgement of principal investigator.

The difference in change from baseline between B. breve and placebo in Frequency of bowel movements after 6 and 12 weeks of supplementation.

Outcome measures

Measure	B. Breve n=42 Participants Capsule containing B breve strain. Participants will be instructed to take 2 capsules of placebo for 4 weeks during the run-in period. On day 1 participants will be instructed to take 2 capsules of B. breve strain in the morning before breakfast for 12 weeks. B. breve: Probiotic capsule. Participants will be instructed to take 2 capsules of placebo for 4 weeks during the run-in period. On day 1 participants will be instructed to take 2 capsules of B. breve strain in the morning before breakfast for 12 weeks.	Placebo n=40 Participants Placebo delivered in capsule format. Participants will be instructed to take 2 capsules of placebo for 4 weeks during the run-in period. On day 1 participants will be instructed to take 2 capsules of placebo in the morning before breakfast for 12 weeks. Placebo: Placebo capsule. Participants will be instructed to take 2 capsules of placebo for 4 weeks during the run-in period. On day 1 participants will be instructed to take 2 capsules of placebo in the morning before breakfast for 12 weeks.	Enterotype 2-B. Breve All subjects (regardless of B. breve or Placebo administration) are divided by enterotype. This sub-arm includes the subjects who share the same Enterotype 2, clustered at the genus level.	Enterotype 2-Placebo All subjects (regardless of B. breve or Placebo administration) are divided by enterotype. This sub-arm includes the subjects who share the same Enterotype 2, clustered at the genus level.	Enterotype 3-B. Breve All subjects (regardless of B. breve or Placebo administration) are divided by enterotype. This sub-arm includes the subjects who share the same Enterotype 3, clustered at the genus level.	Enterotype 3-Placebo All subjects (regardless of B. breve or Placebo administration) are divided by enterotype. This sub-arm includes the subjects who share the same Enterotype 3, clustered at the genus level.
Frequency of Bowel Movements 6 weeks from baseline	-0.36 bowel movements Standard Deviation 2.03	0.82 bowel movements Standard Deviation 2.68	—	—	—	—
Frequency of Bowel Movements 12 weeks from baseline	-0.12 bowel movements Standard Deviation 2.05	0.62 bowel movements Standard Deviation 2.37	—	—	—	—

Adverse Events

B. Breve

Serious events: 0 serious events

Other events: 36 other events

Deaths: 0 deaths

Placebo

Serious events: 0 serious events

Other events: 32 other events

Deaths: 0 deaths

Serious adverse events

Adverse event data not reported

Other adverse events

Measure	B. Breve n=48 participants at risk Capsule containing B breve strain. Participants will be instructed to take 2 capsules of placebo for 4 weeks during the run-in period. On day 1 participants will be instructed to take 2 capsules of B. breve strain in the morning before breakfast for 12 weeks. B. breve strain: Probiotic capsule. Participants will be instructed to take 2 capsules of placebo for 4 weeks during the run-in period. On day 1 participants will be instructed to take 2 capsules of B. breve strain in the morning before breakfast for 12 weeks.	Placebo n=47 participants at risk Placebo delivered in capsule format. Participants will be instructed to take 2 capsules of placebo for 4 weeks during the run-in period. On day 1 participants will be instructed to take 2 capsules of placebo in the morning before breakfast for 12 weeks. Placebo: Placebo capsule. Participants will be instructed to take 2 capsules of placebo for 4 weeks during the run-in period. On day 1 participants will be instructed to take 2 capsules of placebo in the morning before breakfast for 12 weeks.
Gastrointestinal disorders Abdominal discomfort	27.1% 13/48 • Number of events 18 • 12 weeks	21.3% 10/47 • Number of events 21 • 12 weeks
Eye disorders Eye inflammation	0.00% 0/48 • 12 weeks	2.1% 1/47 • Number of events 1 • 12 weeks
Infections and infestations Nasopharyngitis	20.8% 10/48 • Number of events 11 • 12 weeks	23.4% 11/47 • Number of events 17 • 12 weeks
Ear and labyrinth disorders Ear pain	2.1% 1/48 • Number of events 1 • 12 weeks	0.00% 0/47 • 12 weeks
Investigations Blood and urine biochemical indicator abnormal	12.5% 6/48 • Number of events 6 • 12 weeks	0.00% 0/47 • 12 weeks
Musculoskeletal and connective tissue disorders Back pain	12.5% 6/48 • Number of events 7 • 12 weeks	25.5% 12/47 • Number of events 23 • 12 weeks
Injury, poisoning and procedural complications Arthropod sting, Head injury	0.00% 0/48 • 12 weeks	4.3% 2/47 • Number of events 2 • 12 weeks
General disorders Pain, Fatigue, Pyrexia	2.1% 1/48 • Number of events 1 • 12 weeks	6.4% 3/47 • Number of events 3 • 12 weeks
Neoplasms benign, malignant and unspecified (incl cysts and polyps) Melanocytic naevus	2.1% 1/48 • Number of events 1 • 12 weeks	0.00% 0/47 • 12 weeks
Nervous system disorders Headache	37.5% 18/48 • Number of events 33 • 12 weeks	40.4% 19/47 • Number of events 47 • 12 weeks
Psychiatric disorders Anxiety	0.00% 0/48 • 12 weeks	2.1% 1/47 • Number of events 1 • 12 weeks
Renal and urinary disorders Dysuria	0.00% 0/48 • 12 weeks	2.1% 1/47 • Number of events 1 • 12 weeks
Reproductive system and breast disorders Dysmenorrhea, Vulvovaginal discomfort	4.2% 2/48 • Number of events 3 • 12 weeks	6.4% 3/47 • Number of events 6 • 12 weeks
Respiratory, thoracic and mediastinal disorders Oropharyngeal pain, Epistaxis	2.1% 1/48 • Number of events 1 • 12 weeks	8.5% 4/47 • Number of events 5 • 12 weeks
Skin and subcutaneous tissue disorders Onychalgia, Nail discoloration	2.1% 1/48 • Number of events 1 • 12 weeks	2.1% 1/47 • Number of events 1 • 12 weeks
Immune system disorders Allergy to arthropod sting	0.00% 0/48 • 12 weeks	2.1% 1/47 • Number of events 1 • 12 weeks

Additional Information

Director of Clinical Trials

Morinaga Milk Industry Co., Ltd.

Phone: +81-46-252-3067

Email: izen-ri138@morinagamilk.co.jp

Results disclosure agreements

• Principal investigator is a sponsor employee
• Publication restrictions are in place