GINSENOSIDE COMPOSITIONS

Abstract

The present invention relates to the use of ginsencsides to regulate gut microbiota and increase the production of beneficial short-chain fatty acids by said gut microbiota.

Claims

1. (canceled)

2. (canceled)

3. A method of: (i) modulating or adjusting gut microbiota; (ii) increasing the concentration of SCFAs in the gut; (iii) increasing the production of SCFAs by the gut microbiota; and/or (iv) increasing SCFAs blood concentration comprising the administration of an effective amount of a composition comprising ginsenosides to a subject in need thereof.

4. The method of claim 3, wherein the method improves or increases cognition/working memory; treats, reduces, prevents, and/or ameliorates fatigue; improves or increases attention/alertness; and/or improves or increases self-assurance.

5. The method of claim 3, wherein the method regulates satiety; reverses obesity-related and/or metabolic syndrome-related gut microbiota dysbiosis treating or preventing treats or prevents gut microbiota dysbiosis-induced cardiovascular diseases and/or cardiometabolic diseases; treats or prevents low grade inflammation; treats or prevents obesity; treats or prevents sleep disorders; treats or prevents neuropsychiatric disorders; treats or prevents neurodegenerative disease; and/or treats or prevents aged-induced cognitive declined attention, alertness and/or mood.

6. The method according to claim 3, wherein the gut microbiota are selected from isobutyrate producing gut microbiota, valerate producing gut microbiota, isovalerate producing gut microbiota, isocaproate producing gut microbiota, acetate producing gut microbiota, propionate producing gut microbiota and/or butyrate producing microbiota.

7. The method according to claim 3, wherein the gut microbiota are selected from Akkermansia genus and/or Lactobacillus genus.

8. The method according to claim 3, wherein the SCFAs are selected from isobutyrate, valerate, isovalerate, isocaproate, acetate, propionate, and/or butyrate.

9. The method according to claim 8, wherein the SCFAs are selected from acetate, propionate, and/or butyrate.

10. (canceled)

11. (canceled)

12. The method according to claim 3, wherein the ginsenosides are obtained from a root of Panax quinquefolius, Panax ginseng, and/or Panax notoginseng.

13. (canceled)

14. (canceled)

15. The method according to claim 3, wherein the composition comprising ginsenosides comprises ginsenosides from about 3% to about 100% by weight.

16. The method according to claim 15, wherein the composition comprising ginsenosides comprises the ginsenosides: Rg1 from about 1% to 4% by weight of total ginsenosides; Re from about 4% to 35% by weight of total ginsenosides; Rb1 from about 40% to 70% by weight of total ginsenosides; Rc from about 5% to 35% by weight of total ginsenosides; Rb2 from about 2% to 15% by weight of total ginsenosides; and/or Rd from about 9% to 30% by weight of total ginsenosides.

17. The method according to claim 3, wherein the composition comprising ginsenosides is administered in the form of: (a) a pharmaceutical or nutraceutical composition and optionally a pharmaceutically acceptable excipient; or (b) a food composition and optionally a food acceptable ingredient.

18. (canceled)

19. The method according to claim 3, wherein the composition comprising ginsenosides is administered in an amount of from about 100 mg/day to about 2000 mg/day.

20. (canceled)

21. (canceled)

22. (canceled)

23. A method of: (i) modulating or adjusting gut microbiota; (ii) increasing the concentration of SCFAs in the gut; (iii) increasing the production of SCFAs by the gut microbiota; and/or (iv) increasing SCFAs blood concentration comprising the administration of an effective amount of a Panax quinquefolius extract to a subject in need thereof.

24. The method of claim 23, wherein the method improves or increases cognition/working memory treats, reduces, prevents, and/or ameliorates fatigue; improves or increases attention/alertness; and/or improves or increases self-assurance.

25. The method of claim 23, wherein the method regulates satiety; reverses obesity-related and/or metabolic syndrome-related gut microbiota dysbiosis; treats or prevents gut microbiota dysbiosis-induced cardiovascular diseases and/or cardiometabolic diseases; treats or prevents low grade inflammation; treats or prevents obesity; treats or prevents sleep disorders; treats or prevents neuropsychiatric disorders; treats or prevents neurodegenerative disease; and/or treats or prevents aged-induced cognitive declined attention, alertness and/or mood.

26. The method according to claim 23, wherein the gut microbiota are selected from isobutyrate producing gut microbiota, valerate producing gut microbiota, isovalerate producing gut microbiota, isocaproate producing gut microbiota, acetate producing gut microbiota, propionate producing gut microbiota, and/or butyrate producing microbiota.

27. The method according to claim 23, wherein the gut microbiota are selected from Akkermansia genus and/or Lactobacillus genus.

28. The method according to claim 23, wherein the SCFAs are selected from isobutyrate, valerate, isovalerate, isocaproate, acetate, propionate, and/or butyrate.

29. The method according to claim 23, wherein the Panax quinquefolius extract comprises ginsenosides from about 3% to about 100% by weight.

30. The method according to claim 29, wherein the Panax quinquefolius extract comprises the ginsenosides: Rg1 from about 1% to 4% by weight of total ginsenosides; Re from about 4% to 35% by weight of total ginsenosides; Rb1 from about 40% to 70% by weight of total ginsenosides; Rc from about 5% to 35% by weight of total ginsenosides; Rb2 from about 2% to 15% by weight of total ginsenosides; and/or Rd from about 9% to 30% by weight of total ginsenosides.

31. The method according to claim 23, wherein the Panax quinquefolius extract is administered in the form of: (a) a pharmaceutical or nutraceutical composition and optionally a pharmaceutically acceptable excipient; or (b) a food composition and optionally a food acceptable ingredient.

32. (canceled)

33. The method according to claim 23, wherein the extract is administered in an amount of from about 100 mg/day to about 2000 mg/day.

34. (canceled)

Description

BRIEF DESCRIPTION OF THE FIGURES

[0242] FIG. 1: study design of the Cereboost chronic study.

[0243] FIG. 2a: After 4 and 6 h, Cereboost improves proportion of correct responses compared to placebo.

[0244] FIG. 2b: After 2 h, Cereboost improves reaction time compared to placebo

[0245] FIG. 3a: After 2, 4 and 6 h, Cereboost improves proportion of correct responses compared to placebo

[0246] FIG. 3b: After 2, 4 and 6 h, Cereboost improves reaction time compared to placebo

[0247] FIG. 4: After 4 h, Cereboost improves proportion of correct responses compared to placebo

[0248] FIG. 5: After 4 h, Cereboost improves reaction time compared to placebo

[0249] FIG. 6: 6A. After 6 h in acute 2 Cereboost improves proportion of correct responses compared to placebo and compared to acute 1. 6B: After 4 h in acute 2 Cereboost improves reaction time compared to placebo and compared to acute 1.

[0250] FIG. 7: Chronic Cereboost intake improves proportion of correct responses compared to placebo

[0251] FIG. 8: Chronic Cereboost intake limits numbers of errors compared to placebo

[0252] FIG. 9a, b and c: Chronic Cereboost intake reduces fatigue before, during and after a cognitively demanding series of tasks

[0253] FIG. 10: Chronic Cereboost intake increase self-assurance.

[0254] FIG. 11: Chronic Cereboost intake increase joviality

[0255] FIG. 12: Cereboost increased acetate levels in both distal and proximalcolon regions

[0256] FIG. 13: Cereboost increased propionate levels in both distal and proximalcolon regions

[0257] FIG. 14: Cereboost increased butyrate levels in both distal and proximalcolon regions

[0258] FIG. 15: Cereboost increased Akkermansia muciniphila level in the distal colon

[0259] FIG. 16: Cereboost increased Lactobacillus level in the distal colon

EXAMPLES

Example 1

Study Design

[0260] The objective of the experiment was to assess the impact of the intake of 200 mg of an American ginseng extract named Cereboost on healthy adults (n=60) on attention/alertness and mood. Mood is defined as a way participants feel at a particular time: energized, self-assured, sad, hostile, shy.

[0261] The American ginseng extract (Cereboost) used in the present study has a total ginsenosides content from about 10% to 12% (HPLC). The concentration of the specific ginsenosides is: Rg1 from 0.1 to 0.4%, R2 from 0.4 to 3.5%, Rf non detectable, Rb1 from 4 to 7%, Rc from 0.5 to 3.5%, Rb2 from 0.2 to 1.5% and Rd from 0.9 to 3% by weight of the extract The extract has no quintozene and has a particle size of <250 micrometres.

[0262] The study design is represented into the FIG. 1.

[0263] Following recruitment to the study, participants (N=60) started a one-week ‘run-in’ phase where they completed a food frequency questionnaire to give a measure of their habitual diet and attended the laboratory for an initial ‘practice’ session of the cognitive task battery. Thereafter, they attended the laboratory for two further test days over a 2 weeks period On the first test day (acute 1, baseline), participants arrived at the laboratory in a fasted state where they received a standard breakfast, followed by a battery of cognitive and mood tasks. Subjects were then administered their allocated intervention and were re-tested on the task battery at two-hourly intervals over a 6 hours period (acute 1, results versus baseline). Before leaving the laboratory, participants were given sufficient capsules to consume 1 capsule/day of their allocated intervention every morning with their breakfast for the next 13 days. After 2 weeks of treatment, subjects returned to the lab and the procedure for test day 1 was repeated—a baseline test session to assess effects of 14 days of treatment on cognition (Chrome results versus baseline acute 1 and baseline score for acute 2), followed by administration of a final dose of their allocated intervention and test sessions at 2, 4 and 6 hours post-dosing to assess the effects of tolerance (acute 2, results versus baseline 2). For all test sessions a computerised test battery was employed to assess effects on cognitive function and mood. The tasks comprised of:

1) Positive and Negative Affect Schedule Now (PANAS-NOW)

[0264] The Positive and Negative Affect Scale (PANAS-N) will be used to examine mood states at the start and end of the cognitive task battery. It is regarded as a reliable measure for non-clinical populations (Crawford, J. R., & Henry, J. D. (2004). The Positive and Negative Affect Schedule (PANAS). Construct validity, measurement properties and normative data in a large non-clinical sample British journal of clinical psychology, 43(3), 245-265.). Participants are asked to rate the extent to which they experienced each out of 20 emotions on a 5-point Likert scale ranging from “very slightly” to “very much”. Half of the presented emotion words concern negative affect (distressed, upset, guilty, ashamed hostile, irritable, nervous, jittery, scared afraid), the other half positive affect (interested, alert, attentive, excited, enthusiastic, inspired, proud, determined, strong, active). The PANAS-X will be used to measure trait mood. Additionally, Fatigue 1 and 2 will be assessed before and after the cognitive session using a Visual Analogue Scale from 1 to 9.

2) Immediate and Delayed Word Recall

[0265] Using the methodology outlined in Scholey et at., (2010) (Scholey, A et al. (2010). Psychopharmacology, 212(3), 345-356), participants will be presented with a sequential list of 15 words, at a rate of 1 word per second. The participant will then have 60 s to type as many of these words as possible, with the resulting score recorded as a percentage of accuracy. Approximately 35 minutes after the immediate word recall task, participants will be allowed 60 seconds to write down as many items they can remember from the immediate word recall test.

3) Corsi Blocks Task

[0266] This task examines visuospatial memory. Nine identical squares are fixed in a random arrangement on a screen. Participants observe spatial sequences of between two and nine blocks. Four versions of each sequence length presented during the task. The task is to reproduce the sequence, immediately after each presentation by pressing the relevant squares on the screen. The dependent variable is the number of blocks pointed out in the correct order. A novel sequence will be presented on each occasion, the order of which will be counterbalanced across participants.

4) Rapid Visual Information Processing Task (RVIP)

[0267] This task will assess attention processes. In this task a series of digits are presented one at a time on the screen, in quick succession at a rate of 100/min. The participant must examine the continuous series for a sequence of three consecutive even or three consecutive odd digits. The participant must respond once they have detected a sequence string by pressing the space bar as quickly as possible. Up to 8 correct target strings will be presented in each minute, and the task will last approximately 6 minutes. The task will be scored for accuracy.

5) Modified Attention Network Task (MANT)

[0268] This task examines execution function, attention and inhibition. In this task, participants have to respond to a centrally presented arrow, pointing to the left or the right by pressing the corresponding key on the keyboard. The central arrow is flanked by arrows that point in the same (congruent) or opposite (incongruent) direction. In order to perform the task effectively, participants have to ignore the flanking arrows. Previous studies have found that participants show larger latencies and more errors on incongruent trials when compared with congruent trials due to the conflicting interference of the incongruently facing arrows. The response latencies to congruent trials reflect processing speed, while the amount of interference during incongruent trials indicates susceptibility to interference.

6) Task Switch Task (TST)

[0269] This task measures executive function and attention. Participants view a circle with 8 equally spaced radii 2 of which form a bold bisecting line. Numbers are chosen randomly from a set of 1-4 & 6-9 and displayed sequentially in a clockwise direction. A response of higher or lower than 5 is made for trials below the bold line, and even or odd for numbers above the line. General measures of accuracy and response time along with specific measures of switching cost for the first trial after each task change are acquired.

[0270] Data has been analysed using Linear Mixed Modelling for each outcome variable, with post-hoc analysis to further investigate any main or interaction effects between variables Using the design outlined above, three comparisons are available to be made:

[0271] 1) Assessment of acute effect of Cereboost treatment comparing performance at baseline on Test Day 1 with performance two, four and six hours post-treatment (Session 1 Vs session 2, 3, and 4);

[0272] 2) Assessment of acute effect of Cereboost treatment after chronic treatment by comparing performance at baseline on Test Day 14 with performance two, four and six hours post-treatment (Session 5 Vs session 5, 7, and 8)

[0273] 3) Assessment of improvement between the acute 1 versus acute 2 by comparing acute performances at the same time during the day without or after chronic treatment (Session 2 Vs session 6; Session 3 Vs session 7; Session 4 Vs session 8)

[0274] 4) Assessment of the effect of repeated Cereboost treatment by comparing performance at baseline on Test Day 1 with performance at baseline on Test Day 14 (i.e after 2 weeks of daily treatment: (Session 1 Vs session 5)

Results

1—Acute 1 Results

[0275] MANT task: Cereboost has been shown an increase of proportion of correct response and an improvement of reaction time in the MANT (FIG. 2a and 2b). Overall, participants who took Cereboost responded faster as well as more accurately compared to placebo, demonstrating higher attention and alertness.

2—Acute 2 Results

[0276] MANT task: Cereboost has been shown an increase of proportion of correct response and an improvement of reaction time in the MANT (FIG. 3a and 3b)

[0277] Overall, participants who took Cereboost responded faster as well as more accurately compared to placebo, demonstrating higher attention and alertness.

CORSI Task

[0278] Cereboost has been shown an increase of proportion of correct response in the CORSI task (FIG. 4) Overall, participants who took Cereboost responded more accurately compared to placebo, demonstrating higher attention and alertness. Interestingly, 4 h correspond to postprandial dips which appears in the placebo group but not after Cereboost intake.

Switch Task

[0279] Cereboost has been shown an increase of Reaction time in the Switch task (FIG. 5)

[0280] Overall, participants who took Cereboost responded faster compared to placebo, demonstrating higher attention and alertness.

3—Acute 1 Vs Acute 2 Results

MANT Task

[0281] From Acute 1 to acute 2 Cereboost has been shown an increase of proportion of correct response and an improvement of reaction time in the MANT (FIG. 6a and 6b)

[0282] Overall, participants who took Cereboost responded faster as well as more accurately compared to placebo, demonstrating higher attention and alertness. These improvements were increased by 14 days of Cereboost pre-treatment which helped to outperformed the cognitive task.

4—Chronic Results

[0283] MANT task: Chronic Cereboost intake has been shown an increase of proportion of correct response in the MANT (FIG. 7)

[0284] Overall, participants who took Cereboost chronically responded more accurately compared to placebo, demonstrating higher attention and alertness.

[0285] RVIP task. Chronic Cereboost intake has been shown to limit the numbers of error in the RVIP task (FIG. 8).

[0286] Overall, participants who took Cereboost chronically responded more accurately compared to placebo, demonstrating higher attention and alertness.

[0287] Fatigue 1 and 2/PANAS X Fatigue tasks: Chronic Cereboost intake has been shown to limit Fatigue before the series of tasks (Fatigue 1: FIG. 9a), during the task (PANAS-X Fatigue, measuring feelings and emotions such as sleepy, tired, sluggish, drowsy, FIG. 9b) and after the tasks (Fatigue 2: FIG. 9C).

[0288] Overall, participants who took Cereboost chronically feel more energized compared to placebo.

[0289] PANAS X Self-assurance task: Chronic Cereboost intake has been shown to increase self-assurance, regrouping within the PANAX-X feelings and emotions such as: proud, strong, confident, bold, fearless, daring (FIG. 10)

[0290] Overall, due to an increase of self-assurance, participants who took Cereboost chronically feel more confident and determinate.

[0291] PANAS X Joviality task: Chronic Cereboost intake has been shown to increase Joviality, regrouping within the PANAX-X feelings and emotions such as: cheerful, happy, joyful, delighted, enthusiastic, excited, lively, energetic (FIG. 11)

[0292] Overall, participants who took Cereboost chronically feel more joyful.

Example 2

[0293] The objective of the experiment was to assess the impact of an American ginseng extract named Cereboost on the gut microbiota.

Materials and Methods

[0294] The American ginseng extract (Cereboost) used in the present study has a total ginsenosides content from about 10% to 12% (HPLC). The concentration of the specific ginsenosides is: Rg1 from 0.1 to 0.4%, R2 from 0.4 to 3.5%, Rf non detectable, Rb1 from 4 to 7%, Rc from 0.5 to 3.5%, Rb2 from 0.2 to 1.5% and Rd from 0.9 to 3% by weight of the extract. The extract has no quintozene and has a particle size of <250 micrometres.

Simulator of the Human Intestinal Microbial Ecosystem (SHIME®)

[0295] The reactor configuration was adapted from the SHIME® (ProDigest and Ghent University, Belgium) as described by Molly et al. (1993) (Molly. K., Woestyne, M. V., & Verstraete, W. (1993). Development of a 5-step multi-chamber reactor as a simulation of the human intestinal microbial ecosystem Applied microbiology and biotechnology, 39(2), 254-258). Each segment of the SHIME consisted of a succession of three reactors simulating the stomach and small intestine, proximal colon (PC) and distal colon (DC), respectively. Inoculum preparation, retention times, pH, temperature settings and reactor feed composition were previously described by Possemiers, S., Verthé, K., Uyttendaele, S., & Verstraete, W. (2004). PCR-DGGE-based quantification of stability of the microbial community in a simulator of the human intestinal microbial ecosystem. FEMS Microbiology Ecology, 49(3), 495-507. Upon inoculation with a fecal sample of a healthy human adult, a two-week stabilization period was initiated to allow the fecal microbial community to differentiate to colon region-specific microbiota. During a subsequent control period, baseline values for microbial activity and composition were established. After the control period, a three-week treatment period was initiated, during which Cereboost was administered.

Microbial Metabolic Activity

[0296] Samples for microbial metabolic activity were collected three times per week from each colon compartment starting from the control phase. Analysis of SCFA levels, including acetate, propionate butyrate and branched SCFA (isobutyrate, isovalerate and isocaproate), was performed as described by De Weirdt, R., Possemiers, S., Vermeulen. G., Moerdijk-Poortvliet, T. C., Boschker, H. T., Verstraete, W., & Van de Wiele, T. (2010). Human faecal microbiota display variable patterns of glycerol metabolism FEMS microbiology ecology, 74(3), 601-611. Lactate concentrations were determined using a commercially available enzymatic assay kit (R-Biopharm, Darmstadt, Germany) according to manufacturer's instructions.

Microbial Community Analysis

[0297] Samples for microbial community analysis were collected once per week from each colon reactor starting from the control phase. DNA was isolated as previously described by Vilchez-Vargas, R., Geffers, R., Suárez-Diez, M., Conte. I., Waliczek. A., Kaser, V. S. & Pleper D. H. (2013). Analysis of the microbial gene landscape and transcriptome for aromatic pollutants and alkane degradation using a novel internally calibrated microarray system. Environmental microbiology, 15(4), 1016-1039, starting from pelleted cells originating from 1 mL luminal sample. Subsequently, quantitative polymerase chain reaction (qPCR) for Akkermansia muciniphila and Lactobacillus spp. was performed on a QuantStudio 5 Real-Time PCR system (Applied Biosystems, Fester City, Calif. USA). Each sample was analysed in technical triplicate and outliers (more than 1 CT difference) were omitted. The qPCR for Akkermansia muciniphila and Lactobacillus was performed using the protocol as described by Collado et al. (2007) (Collado, M. C., Derrien. M., Isolauri, E., de Vos, W. M., & Salminen, S. (2007).

[0298] Intestinal integrity and Akkermansia muciniphila, a mucin-degrading member of the intestinal microbiota present in infants, adults, and the elderly Appl. Environ. Microbiol. 73(23), 7767-7770) and Furet et al. (2009) (Furet, J. P., Firmesse, O., Gourmelon, M., Bridonneau, C., Tap. J., Mondot, S., . . . & Corthier. G. (2009). Comparative assessment of human and farm animal faecal microbiota using real-time quantitative PCR. FEMS microbiology ecology, 68(3), 351-362), respectively.

Results and Discussion

[0299] As required during the control period SCFA and microbiota composition were all very stable. This indicated that the SHIME model was operated under its most optimal conditions This stability is a prerequisite to make firm statements that effects observed during the treatment truly result from the administered test products.

[0300] Upon initiating the treatment with Cereboost, it was observed that base consumption in the proximal colon mildly increased at the start of the treatment, indicating a stimulation of microbial fermentation This was accompanied by mild increases in gas production. White base consumption and gas production are only a rough indication of microbial fermentation. SCFA measurements provide more detailed insights in the fermentation processes.

[0301] Cereboost significantly increased acetate, propionate and butyrate levels towards the end of the treatment period in both colon regions (FIGS. 12, 13 and 14).

[0302] Acetate (FIG. 12) can be produced by a wide range of gut microbes including among many others Bacteroides spp. (phylum Bacteroidetes) and Bifidobacteria. It followed that the test product significantly increased acetate levels in the proximal colon towards the end of the treatment period.

[0303] An average increase of 7.8 mM (or +42.0%) was observed. In the distal colon, similar effects as in the proximal colon were observed. Again, a strong average increase in acetate levels was observed upon treatment with Cereboost, i.e. an average increase of 6.2 mM (or +21.2%).

[0304] Propionate (FIG. 13) can be produced by a wide range of gut microbes, with the most abundant propionate producers being Bacteroides spp. (phylum Bacteroidetes), Veillonella (phylum Firmicutes) and Akkermansia muciniphila (phylum Verrucomicrobia). It followed that the treatment with Cereboost resulted in significantly increased propionate levels in the proximal colon, i.e. an average increase of +1.77 mM (or +37.9% relative to the control period). In the distal colon, the Cereboost treatment also resulted in a significant increase in propionate levels, with an average increase of 1.96 mM (or +24.1%).

[0305] Butyrate (FIG. 14) is produced by members of the Clostridium clusters IV and XIVa (phylum Firmicutes). In a process referred to as cross-feeding, these microbes convert acetate and/or lactate (along with other substrates) to the health-related butyrate. It followed that Cereboost significantly increased butyrate levels in the proximal colon, resulting in an average increase of 3.0 mM (or +22.6%). In the distal colon, supplementation of Cereboost resulted in significantly increased butyrate levels, i.e an average increase of 1.1 mM (or+10.5%).

[0306] Akkermansia muciniphila indeed remained below the detection limit in the proximal colon in the current study. In the distal colon, Akkermansia muciniphila levels significantly increased upon treatment with Cereboost. This indicates that Akkermansia muciniphila was probably (at least partly) responsible for the increased acetate and propionate concentrations observed in the distal colon upon treatment (FIG. 15).

[0307] Lactobacilli is regarded as beneficial saccharolytic bacteria which is capable of producing high concentrations of lactate. Lactate is an important metabolite in the human colon environment because of its antimicrobial properties, but also because it is the driver of a series of trophic interactions with other bacteria, resulting in the production of downstream metabolites With respect to Lactobacillus levels (FIG. 16), it followed that levels remained unaffected upon treatment with Cereboost in the proximal colon. In the distal colon, Cereboost supplementation increases Lactobacillus levels for the luminal and reached significantly increased in the mucosal compartment (FIG. 16)

[0308] As required during the control period, acid/base consumption, SCFA, lactate, ammonium and microbiota composition were all very stable within and reproducible between each of the SHIME units.

[0309] This indicated that the SHIME model was operated under its most optimal conditions resulting in a stable and reproducible colon microbiota. This stability is a prerequisite to make firm statements that effects observed during the treatment truly result from the administered test products. Upon initiating the treatment with Cereboost, it was observed that base consumption in the proximal colon mildly increased at the start of the treatment, indicating a stimulation of microbial fermentation. This was accompanied by mild increases in gas production. While base consumption and gas production are only a rough indication of microbial fermentation, SCFA measurements provide more detailed insights in the fermentation processes. It followed that Cereboost significantly increased acetate, propionate and butyrate levels towards the end of the treatment period in both colon regions. In terms of acetate and propionate production in the distal colon, this could be linked with significantly increased levels of the acetate and propionate-producing, mucin-degrading Akkermansia muciniphila in the distal colon. Further, it followed that lactate levels remained tow during the course of the treatment period in the proximal colon, indicating proper cross-feeding to other metabolites. In the distal colon on the other hand significantly increased lactate were observed towards the end of the treatment period, which could be linked with elevated Lactobacillus levels. Finally, with respect to markers of proteolytic fermentation, only minor effects were observed in response to the treatment with the different test ingredients.