Dietary fibre formulations

Abstract

A method of treating a person suffering from Irritable Bowel Syndrome (IBS) and other gut-related conditions which involves providing a source of fibre while not aggravating the IBS symptoms in the person, where the method comprises: administering to the person a dietary fibre formulation comprising a ratio of approximately 3 parts high amylose maize starch (HAMS) to 1 part psyllium; wherein the person is administered increasing volumes of the formulation for at least two weeks in an initial ramp-up period; wherein the psyllium protects against fermentation of the HAMS as the formulation passes through the proximal colon, and wherein the HAMS can be fermented in the distal colon of the person to produce butyrate and faeces; and wherein the symptoms of IBS in the person are not substantially aggravated by administration of the formulation.

Description

Dietary Fibre FormulationsField of the Invention

[0001] The invention relates to dietary fibre formulations and dosage regimen for persons that suffer from Irritable Bowel Syndrome (IBS) and other gut-related conditions that provides a source of fibre while not aggravating the IBS symptoms in the persons.Background

[0002] The following discussion of the background art is intended to facilitate an understanding of the present invention only. It should be appreciated that the discussion is not an acknowledgement or admission that any of the material referred to was part of the common general knowledge as at the priority date of the application.

[0003] With their relatively safe and inexpensive characteristics, dietary fibres can be widely applied to improve symptoms for people with IBS. However, it is typically difficult for individuals with IBS to find fibre-rich foods that are potentially low in FODMAPs (fermentable oligosaccharides, disaccharides, monosaccharides and polyols), or those they can tolerate well in order to avoid worsening symptoms.

[0004] Therefore, it would be favourable and cost-effective for individuals with IBS if they could manage the disorder by supplementing their usual diet with a combination of certain isolated fibres that optimise their gut microenvironment and gut function rather than introducing foods which exacerbate their symptoms. This could also ensure they maintain overall diet quality whilst overcoming potential challenges and minimising the possible detrimental effects of following a low FODMAP diet.

[0005] There is evidence in the literature that a combination of isolated fibres as a dietary supplement, may be an option for IBS patients to improve their gut health (So et al., 2022) and a number have been trialed. However, in IBS patients, short-chain carbohydrates resist digestion in the small intestine and are rapidly fermentable in the proximal colon, where they typically result in aggravated gastrointestinal symptoms.

[0006] Thus, there is still a need for sufferers of IBS and other similar conditions that aggravate gastrointestinal systems to obtain alternative sources of fibre.Summary of Invention

[0007] In a first aspect, the invention provides a dietary fibre formulation for use, or when used, in the treatment of IBS, the formulation comprising: one soluble highly fermentable dietary fibre; and one soluble gel-forming partly fermentable dietary fibre also containing insoluble fibre.

[0008] In a preferred embodiment, the one soluble dietary fibre is High Amylose Maize Starch (HAMS). The HAMS preferably comprises approximately 40% resistant starch (RS).

[0009] In a preferred embodiment, the other soluble gel-forming partly fermentable fibre (psyllium). The psyllium preferably comprises powdered psyllium. The psyllium preferably comprises approximately 70% fibre w / w.

[0010] In a preferred embodiment, the formulation comprises an approximately 3:1 ratio of approximately 3 parts one soluble highly fermentable dietary fibre, to approximately 1 part of- soluble gel-forming partly fermentable dietary fibre.

[0011] In a preferred embodiment, the formulation comprises an approximately 3:1 ratio of approximately 3 parts HAMS, to approximately 1 part psyllium.

[0012] In an embodiment, the formulation comprises additional components. The additional components may comprise additives. In an embodiment, the additives comprise flavours, for example, from freeze-dried fruits and vegetables. Such naturally sourced flavours from freeze-dried fruits and vegetables can provide additional beneficial phytochemicals and phenolic compounds to the person consuming the formulation.

[0013] In a second aspect, the invention provides a method of treating a person suffering from IBS or another condition comprising aggravated gastrointestinal symptoms, the method comprising: administering to the person a dietary fibre formulation as described herein.

[0014] In a third aspect, the invention provides a method of treating a person suffering from Irritable Bowel Syndrome (IBS), the method comprising: administering to the person a dietary fibre formulation comprising a ratio of approximately 3 parts high amylose maize starch (HAMS) to 1 part psyllium;wherein the person is administered increasing volumes of the formulation for at least two weeks in an initial ramp-up period; wherein the psyllium protects against fermentation of the HAMS as the formulation passes through the proximal colon, and wherein the HAMS can be fermented in the distal colon of the person to produce butyrate to promote gut health; and wherein the symptoms of IBS in the person are not substantially aggravated by administration of the formulation.

[0015] In a fourth aspect, the invention provides a use of a dietary fibre formulation as described herein for not exacerbating, or alleviating the symptoms of a person suffering from IBS or another condition comprising aggravated gastrointestinal symptoms.

[0016] In an embodiment of the method of the second aspect or third aspect, or use of the fourth aspect, the dosage of the dietary fibre formulation administered to the person increases during an initial ramp up period. The initial ramp up period preferably comprises approximately 11 days.

[0017] In an embodiment, the initial ramp up period comprises two days of approximately 5 g dose of dietary fibre formulation.

[0018] In an embodiment, the initial ramp up period comprises two days of approximately 5 g / day dose of dietary fibre formulation followed by two days of approximately 10 g / day dose of dietary fibre formulation.

[0019] In an embodiment, the initial ramp up period comprises two days of approximately 5 g / day dose of dietary fibre formulation followed by two days of approximately 10 g / day dose of dietary fibre formulation, followed by three days of approximately 20 g / day dose of dietary fibre formulation and another three days of approximately 30 g / day dose of dietary fibre formulation.

[0020] In an embodiment, the initial ramp up period comprises two days of approximately 5 g / day dose of dietary fibre formulation followed by two days of approximately 10 g / day dose of dietary fibre formulation, followed by three days of approximately 20 g / day dose of dietary fibre formulation and another three days of approximately 30 g / day dose of dietary fibre formulation, followed by 11 days of approximately 40 g / day dose of dietary fibre formulation.

[0021] In an embodiment, the ramp up period of increasing amounts of dietary fibre formulation enables the gut microbiome of the person to adjust to the increase in total dietary fibre and fermentable fibres.

[0022] In an embodiment, following the initial ramp up period is a period of maintenance comprising a dose of between approximately 20 g / day to 30 g / day dose of dietary fibre formulation.

[0023] In an embodiment, the dose is divided into more than one administration per day.

[0024] In an embodiment, approximately half of the dose is administered twice a day, or approximately a third of the dose is administered three times a day.

[0025] In an embodiment, approximately half of the dose is administered with a morning meal, and the other approximately half of the dose is administered with an evening meal.

[0026] In an embodiment, approximately a third of the dose is administered with a morning meal, approximately a third of the dose is administered with a midday meal, and approximately a third of the dose is administered with an evening meal.

[0027] In an embodiment, the concentration of psyllium in a daily dose comprises between approximately 1 - 20 g / day.

[0028] In an embodiment, the concentration of HAMS in a daily dose comprises between approximately 10 - 39 g / day.

[0029] In an embodiment, the formulation can be administered as a dietary supplement.

[0030] In an embodiment, the formulation can be administered in combination with one or more food products.

[0031] In an embodiment, the one or more food products may be selected from the group comprising: yoghurt, chia pudding blend, milk from one or more sources, muesli, raw balls, ‘food ink’ for 3D food printing, smoothies, and water-based, and fruit and vegetable juicebased drinks, amongst others.

[0032] In an embodiment, the persons comprise athletes.

[0033] In an embodiment, the persons comprise those taking the formulation for weight loss.

[0034] In an embodiment, the persons comprise those taking the formulation as a preventative for colorectal cancer due to increasing production of butyrate in the colon of the person.

[0035] In an embodiment, the persons comprise those taking the formulation for gut repair and where medication use negatively affects the persons microbiome in their colon or gut.

[0036] In an embodiment, the psyllium prevents fermentation of the HAMS as the formulation passes through the proximal colon of the person.

[0037] In an embodiment, the HAMS can be fermented in the distal colon of the person to produce butyrate to promote gut health.

[0038] In an embodiment, the gastrointestinal symptoms in the person are not substantially aggravated.

[0039] In IBS patients, short-chain carbohydrates resist digestion in the small intestine but then as they progress to the proximal colon, they are rapidly fermented which aggravates gastrointestinal symptoms in the patients.

[0040] Inventors identified a formulation comprising a specific ratio of the soluble highly fermentable dietary fibre High Amylose Maize Starch (HAMS), with the soluble gel-forming partly fermentable dietary fibre psyllium, that when administered to an IBS patient in a ramped dosage regime, enables the gut microbiome of the IBS patient to adjust to the increase in total dietary fibre and fermentable fibres. Importantly, the formulation was also found to shift fermentation of the HAMS to the distal colon which increased production of butyrate to promote gut health in this region while avoiding aggravation of gastrointestinal symptoms. Alternatively, other fibres cause a push the other way to greater fermentation of fibre in the proximal colon which causes increased issues for IBS sufferers.

[0041] More specifically, the process of ramping allows the HAMS-fermenting specific bacteria to increase their numbers gradually, in tandem with the ever-increasing volumes of HAMS. Without this ramping, there would be insufficient numbers of HAMS-fermenting bacteria present in the distal colon resulting in lower butyrate that could cause issues including constipation for the person, further aggravating their IBS symptoms. Thus, the formulation of the invention and initial dosage ramping positively affects the bacterial flora and their growth along the gastrointestinal tract of the person. It also allows an increasing tolerance to the effects of the fermentation of the fibres to minimise symptoms for IBS sufferers as they adjust to fermentation of fibres.

[0042] The shifting of fermentation is provided by the smaller proportion of psyllium in the formulation which has the characteristics of slow fermentability, strong gel-forming, and water-holding capacity, which traps the soluble HAMS granules protecting against fermentation as it passes through the proximal colon. Thereby the HAMS endures to the distal colon for fermentation and butyrate production. Importantly, the 3:1 ratio was found during trials to be required to provide this effect.

[0043] The formulation comprising HAMS and psyllium has not been shown in IBS patients until now and the benefits observed by the inventors include favourably altered gut microbiota, by enriching relative abundances of butyrate-producing taxa, and potentially shifting gut fermentation away from using protein toward fibre as a substrate for colonic health. The mixed fibres also resulted in lower energy and protein intake, and improved Gl- specific anxiety, including sexual subscale of quality of life. However, it did not increase gastrointestinal symptoms and initial quality sleep and dietary patterns were maintained. The inventors showed that the invention highlights that a specific mix and proportion of fibres can improve gut microbiota and Gl-specific anxiety, without impacts on sleep and symptoms in IBS patients on the low FODMAP diet.Brief Description of FiguresFigure 1. Tables of (A) a first dosage regime and (B) a second dosage regime of the dietary fibre formulation of the invention.Figure 2. Table showing demographic information by group.Figure 3. Table showing results of IBS symptom questionnaires by group and timepoint.Figure 4. Table showing summary of variables explained significant individual proportion(%) of total gut microbial variation at three taxonomic levels at T2.Figure s. Table showing results of anthropometry and physical activity by group and timepoint.Figure 6. Table showing daily Gl symptoms by group and phase.Figure 7. Table showing bowel habits and faeces samples by group and timepoint.Figure 8. Table showing daily intake of dietary micronutrients, FODMAP, and five food groups by group and timepoint (Column title with *indicates values from bothdiet + supplement; values in bold indicate significance, with no FDR significance founded).Figure 9. Table showing the relative abundance and fold change of 26 genera by group and timepoint, and number of participants with the direction of change observed in the group at T2.Figure 10. Table showing relative abundance of genera with significant difference between groups and within the group.Figure 11. Table showing the relative abundance and fold change of 42 ASV by group and timepoint, and number of participants with the direction of change observed in the group at T2.Figure 12. Table showing gut alpha diversity indices by group and timepoint.Figure 13. Table showing SCFA concentration and excretion amount comparison by group and timepoint.Figure 14. Table showing SCFA concentration and excretion amount, and faecal weight in the two subgroups of the Fibre-fix group by timepoint.Figure 15. Table showing SCFA concentration and excretion amount, and faecal weight in the two subgroups of the placebo group by timepoint.Figure 16. Table showing serum biomarkers by group and timepoint.Figure 17. Table showing sleep questionnaire results by group and timepoint.Figure 18. Table showing results of mental health questionnaires by group and timepoint.Figure 19. Table showing results of quality of life questionnaires by group and timepoint.Description of Preferred Embodiments

[0044] In order to provide a more precise understanding of the matter of the invention, features of the invention will now be discussed with reference to the following preferred embodiment or embodiments.

[0045] In a preferred embodiment, the dietary fibre formulation of the invention comprises one soluble dietary fibre comprising High Amylose Maize Starch (HAMS), and one soluble gel-forming partly fermentable dietary fibre comprising psyllium.

[0046] In healthy individuals 25 g / day of HAMS is insufficient to increase faecal butyrate levels and change the gut microbiome. However, to initiate a change in a dysbiotic gut, such as those of IBS sufferers, a lower dose is required and the minimum amount of HAMS is approximately 10 g / day. The maximum amount of HAMS is approximately 70 g / day, though this may lower in circumstances as is will also depend on what the dietary fibre formulation of the invention is consumed with and the form it is in. Within the dietary fibre formulation of the invention, the range for HAMS per day is preferably between approximately 10 g to 70 g as some individuals can tolerate more than 15 g / day of psyllium.

[0047] Psyllium husk is tolerable up to a total of approximately 15 g / day for adults who do not have gastrointestinal disease, and a therapeutic amount at 7 g / day to reduce the risk of coronary heart disease (Mysonhimer & Holscher, 2022). Within the dietary fibre formulation of the invention, the range for psyllium per day is preferably between approximately 3 g to 24 g as some individuals can tolerate more than 15 g / day of psyllium.

[0048] The HAMS comprises approximately 40% Resistant Starch (RS) according to nutritional data of HI-MAIZE® 260 produced by Ingredion Incorporated.

[0049] The psyllium is unflavoured powdered psyllium husks (Myprotein®, The Hut Group) and comprises 70.5% fibre according to the nutritional information.

[0050] In the preferred embodiment, the dietary fibre formulation is combined by mixing in an approximately 3:1 ratio with three parts HAMS to one part psyllium.

[0051] A key part of the effect of the dietary fibre formulation is the gradual ramping of the dosage over a 3-week period to allow the gut microbiome time to accommodate to the increase in total dietary fibre and fermentable fibres.

[0052] The initial ramping is outlined in a first dosage regime of the invention shown in Figure 1 , wherein for the first two days, a person is administered 5 g of the formulation each morning with a morning meal for a total of 5 g / day.

[0053] For the next two days, the person is administered 5 g of the formulation each morning with a meal and evening with a meal for a total of 10 g / day.

[0054] For the next three days, the person is administered 10 g of the formulation each morning with a meal and evening with a meal for a total of 20 g / day to end the first week of treatment.

[0055] For the next three days (Day 8 - Day 10), the person is administrated 20 g of the formulation each morning with a meal and 10 g with an evening meal for a total of 30 g / day.

[0056] For the next 11 days the person is administered 20 g of the formulation each morning with a meal and evening with a meal for a total of 40 g / day to complete the three weeks of ramping of the dosage.

[0057] Following the three weeks of ramping of the dosage, the person is administered 10 g of the formulation each morning with a meal and evening with a meal for a total of 20 g / day in a maintenance dosage regime.

[0058] In one alternative embodiment, the formulation may be taken in one daily dose in the total amounts shown in Figure 1 instead of split doses at different times of the day.

[0059] In another alternative embodiment, the formulation may be taken in more than two daily doses, for example, as shown in a second dosage regime of the invention shown in Figure 1 B with the doses split into three daily doses of differing amounts at different times.

[0060] In this respect, the initial ramping is outlined in Figure 1 B, wherein for the first two days, a person is administered 5 g of the formulation each morning with a morning meal for a total of 5 g / day.

[0061] For the next day (Day 3), the person is administered 5 g of the formulation in the morning with a meal and evening with a meal for a total of 10 g / day.

[0062] For the next day (Day 4), the person is administered 5 g of the formulation in the morning with a meal, at noon with a meal, and evening with a meal for a total of 15 g / day.

[0063] For the next three days (Day 5 - Day 7), the person is administered 10 g of the formulation each morning with a meal, 5 g of the formulation each noon with a meal, and 5 g of the formulation each evening with a meal for a total of 20 g / day.

[0064] For the next three days (Day 8 - Day 10), the person is administered 10 g of the formulation each morning with a meal, 10 g of the formulation each noon with a meal, and 10 g of the formulation each evening with a meal for a total of 30 g / day.

[0065] For the next 11 days (Day 11 - Day 21 ), the person is administered 20 g of the formulation each morning with a meal, 10 g of the formulation each noon with a meal, and 10 g of the formulation each evening with a meal for a total of 40 g / day.

[0066] Following the initial three weeks, the person is administered 10 g of the formulation each morning with a meal, 10 g of the formulation each noon with a meal, and 10 g evening with a meal for a total of 30 g / day in a maintenance dosage regime.

[0067] While determined for IBS sufferers, the stated approximately 3:1 ratio with three parts HAMS to one part psyllium and the amounts used in the formulation between the two fibres could be altered for other applications where there is evidence that it may be required.

[0068] The ratio of the dietary fibres in the dietary fibre formulation of the invention may also be varied depending on the type of food it is added to. The ratio for the initial product will be 3:1 (HAMS: Psyllium) but because of the viscosity of the psyllium and the “starchy” taste of HAMS, it may be useful to alter the ratio for different situations and the possible range can be from 1 part / g psyllium to 10 parts / g HAMS and / or 10 parts / g psyllium to 1 part / g HAMS.

[0069] The dietary fibre formulation of the invention may comprise additional components to the one soluble dietary fibre (e.g. HAMS); and one soluble gel-forming partly fermentable fibre (e.g. psyllium).

[0070] In an embodiment, the dietary fibre formulation of the invention may be combined with additional ingredients. In a preferred embodiment, an additional ingredient comprises lupin, other legumes, seeds, and grains. In another embodiment, an additional ingredient comprises one or more selected from the group comprising: grains, seeds, legumes natural and synthetic flavours, natural and synthetic colourings. The grains may comprise any one or more comprising whole grains, refined grains, enriched grains, or fortified grains. The grains may comprise any one or more comprising barley, bulgur, farro, millet, quinoa, black rice, brown rice, red rice, wild rice, oatmeal, popcorn, whole-wheat flour, whole grain breakfast cereals.

[0071] In some non-limiting examples, the additional components may comprise, amongst others, flavours, for example flavours from freeze-dried fruits and vegetables. Such flavours from natural sources can provide additional benefit to support gut health in terms of additional phytochemicals and phenolic compounds, thereby increasing the nutritional value of the formulation.

[0072] The dietary fibre formulation of the invention can be consumed with a meal as an individual product or within a meal. Within a meal may comprise adding the dietary fibre formulation to a food or drink product.

[0073] Some non-limiting examples of food products that could include or be mixed or combined with the dietary fibre formulation, or used as an ingredient in other food products, comprise, amongst others: A yoghurt blend similar to “Chia pudding” with added ingredients comprising but not limited to, chia seeds, nuts, LSA, dried fruit, honey, fruit, dried fruit powders; a milk (oat, rice, soy, almond, cow, goat) blend similar to blancmange with added ingredients comprising but not limited to, chia seeds, nuts, LSA, dried fruit, honey, fruit, dried fruit powders; bircher muesli; muesli bar; raw balls; “Food Ink” for 3D food printing; drinks including milk and yoghurt-based smoothies (oat, rice, soy, almond, cow, goat); and drinks including water-based and fruit and vegetable juice-based drinks.

[0074] The dietary fibre formulation of the invention may also be provided as a dietary supplement.

[0075] The dietary fibre formulation of the invention is directed towards administration and use by IBS sufferers and the treatment thereof as identified from clinical trials.

[0076] Clinical Trial Data

[0077] Methods - Participants and Sample Size

[0078] Inclusion and exclusion criteria have been included in the protocol paper (Yan et al., 2020) and are detailed in the supplementary material. In summary, adult IBS patients (18-65 years old) were included if they had stable symptoms (self-reported), a clinal diagnosis of IBS by a gastroenterologist or other medical professional, and been on a low FODMAP diet for at least one month prior to the intervention. Participants were excluded if they were current smokers or had a known diagnosis of other gastrointestinal illness.

[0079] A sample size calculation was performed using G*Power version 3.1 .9.2 (Faul et al. 2007). The calculation was based on a within-between interaction design with an expected effect size as 0.3 (a change in log SCFA concentration) with 80% power and an a-error at 5%, whereby a minimum sample size of 24 was required. Accounting for a 15% attrition rate, the final minimum required sample size included 28 participants.

[0080] Methods - Trial Protocol

[0081] The study protocol has been published elsewhere (Yan et al., 2020). In summary, this study was designed as a randomised, double-blind, placebo-control trial, including a one- week baseline and a three-week intervention. At baseline (Timepoint 1 , T1 ), participants were assessed for anthropometric measures, such as body weight (kg), waist and hip circumference (cm) and percentage of lean and fat mass (determined by BOD POD, an Air Displacement Plethysmograph). Their fasting blood sample were collected to test the blood biomarkers of inflammation including lnterleukin-6 (IL-6), IL-1 [3, Tumour Necrosis Factor alpha (TNF-a) and high-sensitivity C-reactive protein (hs-CRP). The baseline concentration and excretion amount of SOFA were examined from a 24-h faecal sample (all faecal output within the 24 hours). A 3-day weighed food diary was recorded by each participants for analysing their habitual dietary intake. The general diet quality was assessed using Aussie- Dietary quality index (Zarrin et al., 2013), detailed in the supplementary material. Varied questionnaires were used to assess IBS symptoms, FODMAP intake, sleep, mental health, quality of life and physical activity, as detailed in the study protocol (Yan et al., 2020). After the baseline week, participants were randomly assigned to one of two groups; either the Fibre-fix supplement or placebo, for a period of three weeks.

[0082] For the purpose of describing the invention, reference herein to ‘Fibre-fix’ also refers to a dietary fibre formulation according to the invention. Fibre-fix™ is also now more commonly referred to as GutFeels™ by the inventors.

[0083] Both groups increased their supplement dose for 11 days (from 5 g to 40 g / d) until the full dose was consumed for a further 10 days (40 g / d), as described in the study protocol (Yan et al., 2020). Education about consumption, calculation of compliance and specimen collection and analysis are detailed in the supplementary material. A wrist-actigraphy monitor, a Readiband™ v5 (Readiband, Fatigue Science Inc., Canada) (validated Dunican et al., 2017), was provided to each participant to capture daily sleep data. Throughout the four weeks, participants also recorded their daily symptoms via an online checklist. At the end of the study (Timepoint 2, T2), the assessments conducted at T 1 were repeated. The study was approved by the Human Research Ethics Committee (HREC) of Edith Cowan University (ECU) (ID: 2019-00619-YAN). The study was registered with the Australia and New Zealand Clinical Trial Registry (ACTRN12620000032954). All participants consented before the commencement of the study.

[0084] Statistical Methods

[0085] The per-protocol analysis was undertaken using data from participants who completed the study, whereby non-compliant participants were excluded. Linear Mixed Modelling (LMM) was used to determine both within- and between-group differences in health outcomes via the group x time interaction effect and the main effects (if the interaction is not significant), with BMI and age included as covariates. Gender was not included due to the small number of male participants (n=3). Post-hoc tests were conducted if a significant effect was detected. The significance level was set at P < 0.05. Furthermore, false discovery rate (FDR) correction was performed on all raw P-values to minimise false positive results, and an effect was noted as marginal when the raw P < 0.05, but the FDR P > 0.05. For gut microbiota, a combination of R, Primer 7 and Permutation Multivariate Analysis of Variance (PERMANOVA) + (PRIMER-E, Plymouth), was used to analyse bacterial community structure at three taxonomic levels; phylum, genus and Amplicon Sequencing Variant (ASV). The relative abundance data were square root transformed and a Bray-Curtis dissimilarity matrix was calculated, followed by the deployment of Principal Coordinates Analysis (PCoA) to visualise findings of the three taxonomic levels at T1 and T2. For determination of significant associations at baseline and longitudinal dissimilarities between the two groups, PERMANOVA and dissimilarity percentage (SIMPER) were conducted. Distance-based linear models (DistLM) and distance-based redundancy analysis (dbRDA) were used to integrate gut microbial findings with all other relevant aspects. Further details of the statistical analyses are provided in the supplementary material.

[0086] Results - Participants

[0087] Overall, 30 participants consented to the study, and 26 participants (87%) completed the study and were included in the analysis (Supplementary results), which fulfilled the minimum required sample size of 24 participants. The mean age of participants was 37 ± 13 years, whereby 88% (n = 23) were female, with no between-group difference in demographic characteristics (Figure 2). Compliance for the placebo group (94.8 ± 5.5%) was significantly higher than the Fibre-fix group (89.0 ± 6.5%) (P = 0.022), and both groups met the a priori compliance criteria (> 80%). No significant change was noted in anthropometries and physical activity at T2 (Figure 5).

[0088] Results - IBS Symptoms and Bowel Habits

[0089] The severity of symptoms was assessed by two questionnaires: the Gastrointestinal Symptom Rating Scale for IBS (GSRS) and the Irritable Bowel Syndrome-Symptom Severity Scale (IBS-SSS). Both groups had a moderate level of symptoms and did not change overthe intervention (Figure 3). Likewise, no significant interaction effect was found in daily Gl symptom scores throughout the 4-week study (Figure 6).

[0090] Whilst the effect of treatments on bowel habits were examined longitudinally, no significant interaction effects were observed for stool weight and type (self-rated using Bristol Stool Chart) or the number of bowel movements (an average of the last three days of each timepoint, same period as the 3-day food diary) (Figure 7).

[0091] Results - Dietary outcomes (FODMAP, Macronutrients)

[0092] At T1 , no significant between-group difference was noted for all dietary outcomes, except for energy from carbohydrate (in percentage), which was higher in the placebo group than the Fibre-fix group (40.6 ± 8.9 % vs 31 .5 ± 7.2%, P = 0.009, FDR-P = 0.045).

[0093] Cross the whole study there was no significant Group x Time interaction effect for any dietary outcomes, including Dietary Quality Index (DQI), or FODMAP intake, where these dietary markers remained stable in the respective groups (Figure 8). However, marginal evidence of an interaction effect was observed for intakes of energy and protein (P = 0.033 and P = 0.004 respectively; both FDR P > 0.05), where both showed lower intakes in the Fibre-fix group at T2* in contrast to increases in the placebo group (Figure 8).

[0094] A marginal interaction effect (P = 0.016, FDR P > 0.05) was observed in habitual dietary fibre derived from diet only, with a decrease in the Fibre-fix group at T2; however, this reduction was offset by the Fibre-fix supplement, leading to an overall increase in fibre intake at a total of 33.1 ± 2.4 g / d (Figure 8), which met daily Australian recommended levels for males (30 g / d) and females (25 g / d). The Fibre-fix group marginally increased their intake of resistant starch to 9.5 ± 0.5 g / d at T2* from their T 1 level at 2.0 ± 0.3 g / d (P < 0.05, FDR P > 0.05) (Figure 8). This reached the average level of Australian adults at 9.5 g / d (Rees et al., 2021), although only half of the suggested amount of 20 g / d (Baghurst et al., 1996).

[0095] Results - Gut Microbiota

[0096] PERMANOVA analysis did not show significant microbial composition changes at the phylum level (P = 0.610). However, overall microbial composition significantly altered at the genus level (P = 0.003), where between-group differences were not significant at T1 (P = 0.262), but were significant at T2 (P = 0.008), despite no Group x Time interaction effect. Similarly, no significant between-group difference at T1 (P = 0.275) was observed at ASVlevel. However, after the intervention, the microbial composition of the two groups significantly differed (P = 0.002).

[0097] The genus of Ruminococcus E significantly increased to a relative abundance of 11.33% at T2 with a fold change (fc) of 5.5 (P < 0.01 ) in the Fibre-fix group, where 92.3% (n=12) of participants recorded an increase (Figure 9). This relative abundance was greater than that of the placebo group at T2 (P < 0.001 , FDR P = 0.0037) (Figure 9). Similarly, Fibrefix enriched Prevotella, fc = 5.9 (P > 0.05), which was significantly more abundant at T2 compared to the placebo group where the relative abundance of Prevotella remained unchanged (Figure 9). A similar but insignificant trend was found in Akkermansia, with fc = 2.6 in the Fibre-fix group compared to a fc = -1.8 in the placebo group (Figure 9). Among all classified genera, a total of three genera significantly changed within the Fibre-fix group, including Ruminococcus E (increased), Anaerobutyricum (decreased) and Catenibacterium (decreased), with none occurring in the placebo group (Figure 10, also showing between- group difference at T1 and T2).

[0098] Within the Fibre-fix group, ASV 14 Ruminococcus was marginally enriched, 1.6% (T1) vs 6.7% (T2), fc = 4.3 (P = 0.010, FDR.P > 0.05) (Figure 11 ). Error! Reference source not found.Other enriched ASVs included ASV 12 Akkermansia muciniphila, ASV 21 Ruminococcus E bromii A sp003526955, ASV 11 Prevotella copri A, ASV 18 Prevotella copri A sp900546535 and with respective fcs of 32.9, 9.7, 8.8 and 9.1 , despite their statistical insignificance (Figure 11 Error! Reference source not found. )Error! Reference source not found..

[0099] Regarding gut diversity indices, the placebo group was observed to have a significantly lower diversity at baseline than the Fibre-fix group, including Chaol index, Shannon index and Fisher index (Figure 12). No significant interaction effects were noted after the intervention, where the indices remained stable in the Fibre-fix group at T2 (Figure 12).

[0100] Results - Faecal SCFA and Blood Biomarkers

[0101] No significant interaction effects were shown on the concentration and excretion amount of SCFA in 24-hour stool samples (Figure 13). However, significant changes in faecal output and SCFA were found, when dividing the Fibre-fix group according to their T1 faecal output lower or greater than 150 g / d, a faecal index associated with improving fermentation products in relation to colon cancer risk (Birkett et al., 1997). At T2, the Fibre-fix LOW subgroup(n = 5) significantly increased the faecal butyrate concentrations and excretion by 51% and 289% of their T1 level (both P , 0.05, FDR P > 0.05), where their mean faecal output also increased from 78 to 162 g / d (detailed in the Supplementary Results and Figure 14 A and Figure 15).

[0102] Blood biomarkers of inflammation status were collected, but most of the results for IL-1 [3 and TNF-a were below detection limits, leading to insufficient sample sizes for further analysis. For IL-6 and hs-CRP, no significant changes were observed (Figure 16).

[0103] Results - Sleep

[0104] Participants exhibited normal sleep duration, sleep latency and sleep efficiency measured daily across the baseline week, according to the National Sleep Foundation's recommendations (Ohayon et al., 2017), whereby no significant change was observed during the intervention in either group in relation to objective sleep measures . A marginal interaction effect was observed for Time at Sleep Onset (P = 0.031 , FDR P > 0.05), where the placebo group delayed their sleep onset time, whilst the Fibre-fix group maintained their sleep onset time throughout the study. Similarly, no significant effects were observed in sleep questionnaires (Figure 17). These sleep outcomes indicated that sleep patterns and behaviours were stable in the Fibre-fix group over the duration of the study.

[0105] Results - Mental Health and Quality of life

[0106] Suggestive evidence of an interaction effect was observed for the Visceral Sensitivity Index (VSI) (P = 0.034, FDR-adjusted P > 0.05), where the Fibre-fix group had a reduction of 7.4 ± 2.6 units at T2, whilst VSI remained steady for the placebo group (Figure 18). No significant interaction effects occurred on measures of the Depression Anxiety Stress Scales (DASS21 ) (Figure 18). Two genera, Alistipes and Coprococcus, showed a significant correlation with VSI score (rp= 0.750 and 0.743, P <0.001 , respectively).

[0107] A marginal interaction effect was found on the sexual subscale of the IBS-quality of life (QOL) (P = 0.019, FDR-adjusted P > 0.05), where the Fibre-fix group had a within-group increase of 12.5 ± 4.9 (P = 0.018), whilst there was no notable change for the placebo group (Figure 19). No significance was noted for all remaining seven subscales of IBS-QOL (Figure 19).

[0108] Results - Gut Microbiota - DistLM

[0109] DistLM analysis showed that seven variables individually explained a significant proportion of the variability of gut microbial composition at the three taxonomic levels of gut microbiota (phylum, genus and ASV) (Figure 4). The Epworth Sleepiness Scale (ESS), indicating the daytime sleepiness contributed significantly at all three levels, together with PSQI at two levels, suggesting the influence of sleep on the gut microbiota composition (Figure 4).[001 10] Supplementary Methods[001 1 1 ] Inclusion and exclusion criteria[001 12] Those who met the inclusion criteria were recruited into the study. These included: men and women between 18 - 65 years old with a clinical diagnosis of IBS, using the Rome IV edition diagnostic criteria (Drossman and Hasler, 2016), diagnosed by a gastroenterologist or other medical professional; those who had been on a low FODMAP diet for at least one month prior to the intervention and had stable symptoms (self-reported); those who were available to attend local clinic visits and were willing to consume the Fibre-fix supplement or matched placebo; and those who had a BMI between 18.5 - 40 kg / m2.[001 13] Participants were excluded if they: were current smokers; were pregnant or planning to become pregnant; had a known diagnosis of other gastrointestinal illness (e.g., Inflammatory Bowel Disease, malabsorption of any macronutrients, bowel resection, coeliac disease); had undergone previous abdominal or gastrointestinal surgeries, had severe mental health and sleep-related conditions (e.g., insomnia); had renal or hepatic diseases, and major medical illness; currently used pharmaceutical agents that could modify or treat IBS (e.g., probiotics, antibiotics, eluxadoline, lubiprostone and linaclotide) or sleep conditions; followed other restrictive dietary patterns or therapies (e.g., low-carbohydrate, or high-protein diets, keto / Paleo-diet); took any prebiotics; had any other diseases, conditions or habits that may interfere with completion of the study.[001 14] In response to the COVID 19 pandemic, which caused difficulty in recruitment, the eligibility criteria relevant to probiotic and antibiotic use was relaxed to include those who were willing to stop taking current probiotics / antibiotics for at least 3 weeks prior to study commencement and cease using them during the study. Accordingly, these eligible participants completed a 3-week wash-out prior to the starting date.[001 15] Diet Quality Index[001 16] Aussie-DQI (a diet quality index for Australian population) (Zarrin et al, 2022) was used for assessing general diet quality, where it has been listed as one of the preferred DQI used in Australian adults according to a systematic review in 2020 (Hlaing-Hlaing, 2020). Aussie-DQI is comprised of 11 components, each scored out of ten points (ten indicating the guideline was sufficiently met), with one component having four sub-components and each was scored out of five points. The detailed components and scoring instruction were adapted from Table 1 of the original Aussie DQI validation paper Zarrin et al (2022), thereby making the total Aussie-DQI score within a range of 0 to 120. Higher Aussie-DQI scores indicate a better diet quality with desirable intake of nutrients, a greater adherence to the Australian Dietary Guidelines (ADG) (NHMRC, 2013), and inversely, is associated with risk of cancer mortality Zarrin et al (2022).[001 17] In 2017, ADG revised the Suggested Dietary T arget to 2,000 mg / d for adults and ‘not determined’ for sodium (NRVAUNZ, 2017). After consultation with authors of the Aussie-DQI, we agreed to adjust the cut-off of sodium to align with the latest recommendation of 2,000 mg / d. This also suggests the results by NRVAUNZ, 2017 are slightly outdated. Despite this, it is mostly correct and provides a valuable baseline reference for the current study.[001 18] Education of consumption and calculation of compliance[001 19] Participants were educated about the best procedure for consuming the supplement. Participants were recommended to take the supplement immediately after mixing it thoroughly with at least 320 ml water or a beverage at room temperature after stirring thoroughly for 20 seconds (before the fibre settles to the bottom). Moreover, participants were instructed to not heat / cook / bake the supplement, or add the mixture to hot drinks above 60 °C (140 °F) to avoid altering fibre content. Participants were also instructed to record the time of consumption via the online daily checklist. If a sachet was missed, participants were instructed to keep consuming sachets as scheduled, and bring back empty packages and missed sachets.

[0120] Both sachet packages and checklist records were used to calculate the compliance. Consumption of greater than 80% of the whole amount of the fibre supplement over the 3- week intervention was considered compliant with the study protocols. Therefore, participants who did not meet the criterion were regarded as non-complaint and were excluded from the per-protocol analysis.

[0121] Blood sample collection

[0122] Venous blood samples were collected at both first and final clinic visits taken by a trained phlebotomist at Edith Cowan University (ECU). Participants were required to fast overnight from 10 pm the night before their morning appointment. Approximately 15 ml of blood was collected in three kinds of vacuum blood collection tubes, 5 ml premium VACUETTE® tube serum gel gold cap, 4 ml premium VACUETTE® tube lithium heparin, and 4 ml premium VACUETTE® tube K3 EDTA, using ECU Standard Operating Procedures for Human Phlebotomy. Following Standard Operating Procedures for Human Blood Centrifuging and Aliquoting, the collected samples were stood to clot for 30 minutes and centrifuged before separating plasma and serum, and then frozen at -80 °C after being aliquoted into 2 ml vials.

[0123] Blood sample analysis

[0124] Analysis of cytokines (High-sensitive lnterleukin-6 (IL-6), IL-1p, tumour necrosis factor-a (TNF-a), and high-sensitive C reaction protein (hs-CRP) was assessed in accordance with the protocols from the National Association of Testing Laboratories by SA pathology laboratory in Adelaide, South Australia. Their assay cytokine test has received National Association of Testing Authorities, Australia (NATA) accreditation. Cytokines, high- sensitive IL-6, IL-1 p, TNF- a, were measured in serum using BD™ Cytometric Bead Array (CBA) Kits (BD Biosciences, Sans Jose, CA, USA) with the addition of the BD™ CBA Enhanced Sensitivity Flex Set, a bead-based immunoassay, being capable to quantitate soluble analytes present in very low concentrations. Following the manufactures’ assay instructions, the BD™ CBA kit was used in conjunction with a BD CBA Human Enhanced Sensitivity Master Buffer Kit, a flow cytometer on BD FACSCanto, and FCAP Array™ Software for data analysis (BD Biosciences, Sans Jose, CA, USA). For the measurement of hs-CRP in serum, particle enhanced immunoturbidimetric assay (Roche Diagnostics, Indianapolis, IN, USA) was used following manufacturers instruction. The lower detection limits of the all tests summarised as followed: IL-1 p: 0.048 pg / mL; IL-6: 0.068 pg / mL; TNF- a: 0.067 pg / mL; hs-CRP: 0.15 mg / L.

[0125] 24-hour stool collection

[0126] Participants were provided with a stool collection kit, including a portable cooler bag, frozen icepacks and an instruction sheet. Participants were instructed to collect the whole stool sample of each bowel movement within the 24-h period. Upon receipt, stool samples were immediately weighed and stored at -80 °C, according to ECU Standard OperationProcedure for stool collection. To generate 24-hour faecal output, all weights of stool samples were summed and the total weight of collecting bags and cable ties was subtracted.

[0127] Faecal SCFA analysis

[0128] Faecal SCFA analysis was undertaken at ECU. The concentration of SCFA was determined by gas chromatography (Stinson et al, 2019). In brief, an acidified aqueous methanol solution was used to extract SCFA from faecal samples, followed by separating of SCFA by gas chromatography with a fatty acid column (Zebron ZB-FFAP 30m x 0.53mm x 1 m pm supplied by PhenomeneX). The SCFA concentrations were determined via internal standards. The SCFA outcome measures included six acids: acetic acid, propionic acid, isobutyric acid, butyric acid, isovaleric acid and valeric acid. These six acids added together to calculate the total SCFA concentration. The acid concentration was measured in pmol / g of wet faeces. The excretion amount of each SCFA type and all SCFA was calculated by multiplying the respective concentration by the participant’s total 24-hour stool weight (gram), and the unit was mmol per 24 hours.

[0129] Gut microbiota analysis

[0130] Microbial analyses were performed at the WA Human Microbiome Collaboration Centre, Curtin University, Western Australia. The analysis of the gut microbiota was similar to the approach in the study of Jones et al (2021 ). In summary, DNA was extracted using the QIAamp PowerFecal Pro DNA kit (Qiagen) using Qiacube extraction platform following PCR inhibition assessment of each DNA extract. PCR-free ligation protocol was thereafter deployed for the process of library building. Microbiome signatures were generated using the Illumina MiSeq platform using uniquely barcoded 16S rRNA gene primers (515-806 (V4)) for quality control, mock communities, extraction controls and non-template control samples were included in the sequence analysis. Sequence read quality was initially assessed with FastQC before demultiplexing and pre-processing by GHAPv2, an in-house tool. Cutadapt (Martin, 2011 ) was used for removal of all non-biological sequences. DADA2 (Callahan et al, 2016) was then used for quality filtering, error correction, amplicon sequence variants (ASVs) picking. A trained naive bayes classifier then assigns ASVs to genus / species against a curated database of microbial reference sequences, Genome Taxonomy Database (Parks et al, 2018). If the microbes were not able to be identified at genus or species level, the Basic Local Alignment Search Tool (BLAST) of National Center for Biotechnology Information (NCBI) will be used to identify via the comparison the nucleotide sequences of the specificASV to the sequence databases. The sequence data yielded a total of 3031 ASVs which were further quality filtered according to the five principles below and in the order:1 ) Each ASV had greater than or equal to 100 reads in total (1304 ASVs included).2) Each ASV was represented in more than 5% (n=3) of the total numbers of samples (1095 included)3) The reads in the Non-Template Control counting for the ASV reads were below 1% (1058 included)4) The reads in the Extraction Control counting for the ASV total read were below 1% (1054 included)5) Any ASV not classified beyond Kingdom (with assigned phylum) was deleted.This resulted in a total of 988 reads.

[0131] Statistical analysis

[0132] For daily sleep measurements, those who had >50% complete data for each phase of the entire study, including a baseline week (7 days), dose ramping-up period (11 days), and full dose period (10 days) were included in the analysis. The total 24-h stool weight (wet weight) was calculated and assessed to be above or below 150 g / d, a faecal index relevant to colon cancer risk (Birkett et al, 1997). Anthropometric data, results of stool SCFA and blood biomarkers, dietary assessment data (FODMAP, nutrients and food groups), scores of all questionnaires, daily objective sleep data collected via Readiband, as well as scores of symptom severity collected from daily checklist were all exported to MS Excel for storage and cleaning. Missing data and outliers were checked for scientific accuracy. The resulting spreadsheets were imported into SPSS v27.0 (IBM, 2013) for analysis.

[0133] All continuous outcome variables were examined for normality using the Shapiro- Wilk test. Descriptive statistics in the form of mean and standard deviation (Std. D) were used to describe numerical variables that were normally distributed, and an independent t-test was conducted to identify the difference between the placebo and intervention group at baseline. Median and interquartile range (IQR) were reported for non-normally distributed data, whereby a Mann-Whitney II test was used instead. For nominal variables, frequencies and proportions were reported; the distribution of categorical variables across each group was tested using the Pearson Chi-square test. False discovery rate (FDR) correction was performed on all raw P-values to minimise false positive results. When unadjusted (raw) P was < 0.05 and FDR-adjusted P > 0.05, it would be noted as marginal significance / evidencesuggesting a trend, whereas FDR-adjusted P < 0.05 would be deemed significant, and with a symbol (or a superscript) marking the P value.

[0134] With regard to gut microbiota, at the levels of phylum, genus and ASV, the relative abundance data were square root transformed and a Bray-Curtis dissimilarity matrix was calculated, followed by the deployment of Principal Coordinates Analysis (PCoA) to visualise findings of the three taxonomic levels at T1 and T2. For the determination of significant associations at baseline and longitudinal dissimilarities between the two groups, PERMANOVA and dissimilarity percentage (SIMPER) were conducted. Distance-based linear models (DistLM) and distance-based redundancy analysis (dbRDA) were used to integrate gut microbial findings with all other relevant aspects, such as: dietary intake, sleep, mental health and quality of life, blood and faecal biomarkers at each of the three levels contributing to 80% of participants' gut microbial composition variations, in order to account for the variability of the gut microbial composition at each level. For the DistLM analysis of the T2 result, one participant was removed due to their incomplete questionnaire. The dbRDA ordination was utilised to visualise variables, which contributed to 80% of the variations of gut microbial composition of participants.

[0135] The Shannon index, Chaol index and Fisher index were three indices used to indicate alpha microbial diversity using rarefied data at both timepoints. These indices, and the observed species, were exported to SPSS v27.0 for LMM analysis to identify any between-group and within-group changes with BMI, and were age adjusted in the model. The significance level was set at P < 0.05.

[0136] Supplementary Results

[0137] Participants

[0138] A rolling recruitment occurred for this research from July 2020 July - July 2021 in Perth, Western Australia. Overall, 30 participants enrolled after signing the online consent form; however, one female participant withdrew during the baseline observation due to pregnancy, and three participants did not complete the study. In the placebo group, one participant was lost to follow-up and another self-reported feeling unwell and withdrew. One female in the Fibre-fix group reported nausea and withdrew. Therefore, 26 participants completed the study and were included in the analysis, which fulfilled the minimum required sample size of 24 participants. Out of all participants, 73% had a diagnosis of IBS beyond one year, whereas 35% had been on a low FODMAP diet for more than one year, and 77%were at the phase of a personalised low FODMAP diet (Figure 2). On average, participants had a healthy weight, level of adiposity and blood pressure, and engaged in physical activity, whereby no significant change was noted in anthropometries and physical activity (Figure 5).

[0139] Faecal SCFA

[0140] Suggestive evidence was noted for a Group x Time interaction (P = 0.042, FDR- adjusted P > 0.05) between the two sub-groups in the Fibre-fix group, the Fibre-fix HIGH (n = 8, T1 faecal output > 150 g / d) and the Fibre-fix LOW (n = 5), where the latter had an increase in the mean faecal output to 161.9 g / d at T2 (no change in the daily number of bowel movements, Figure 14). Individually, four of the five participants in the Fibre-fix LOW subgroup had the faecal output above 150 g / d at T2, whilst only one third of participants had this response in the placebo LOW subgroup. For SCFA, marginal evidence of interaction effects was noted in the excretion amount of total SCFA, acetate and butyrate, as well as butyrate concentration, where the Fibre-fix LOW subgroup had significant increases (except for total SCFA) in comparison to its T 1 level and the Fibre-fix HIGH group did not significantly change (Figure 14). Within the placebo group, marginal evidence of interaction effects was observed for faecal output (P = 0.036, FDR-adjusted P > 0.05), where the Placebo HIGH (n = 8, T1 faecal output > 150 g / d) had a significant reduction by -44.1% (P = 0.026) (Figure 15).

[0141] In the Fibre-fix group, the mean score of gas increased from 3.7 at Phase 1 and 4.3 at Phase 2, and then declined to 3.9 at Phase 3, without statistical significance noted (Figure 6). This aligned with anecdotal self-report of participants taking Fibre-fix that they experienced bloating / more gas during the first few days (up to one week at most), which then was shown to revert to usual patterns. The score of gas was stable, 3.6, 3.4 and 3.7, respectively throughout the study in the placebo group (Figure 6).

[0142] For the result of Food Groups intake, grain and refined grain showed marginal interaction effects (P = 0.004 and 0.018, respectively, both FDR-adjusted P > 0.05) (Figure 8). The placebo group experienced significant increases in their grain intake, while it did not significantly change in the Fibre-fix group at T2 or T2*. In contrast, for the serve of protein foods, namely meat and meat alternatives (interaction P = 0.038, FDR adjusted P > 0.05), this was reduced in the Fibre-fix group by 0.8 ± 0.3 serve / d from T1 to 2.5 serves / d at T2*, whereas it was maintained in the placebo group, which is aligned with changes in protein intake in grams (Figure 8).

[0143] However, the dietary fibre formulation of the invention may also be used by athletes and sports people. The inventors have case studies showing the effect of the formulation in normalising / stabilising the athlete’s gut.

[0144] The dietary fibre formulation of the invention may also be used in weight loss. In the inventor’s clinical trial in IBS patients, after only 3 weeks they observed lower energy intake in the group consuming the formulation.

[0145] The dietary fibre formulation of the invention may also be used in a method or use for preventing colorectal cancer (CRC). The formulation shifts fermentation and increases production of butyrate more distally in the colon. The development of CRC can be associated with a lack of butyrate in the distal colon as butyrate regenerates the colon preventing DNA damage and promoting apoptosis of damaged cells.

[0146] The dietary fibre formulation of the invention may also be used in a method or use for treating other diseases and conditions of the gastrointestinal system of a human. As the formulation selects bacteria that promote butyrate production more distally in the colon, this product may be beneficial in relieving or treating other conditions of the gastrointestinal system of a human.

[0147] The dietary fibre formulation of the invention may also be used in a method or use for repairing a person’s gut, colon, and gastrointestinal system. As this product assists to promote growth of bacteria that increase butyrate production more distally in the colon, this product may repair the gut, and which would be particularly beneficial to those individuals who eat a low fibre diet (namely highly processed diet of refined carbohydrates, low in vegetables and fruit). The repair process may allow those with IBS or other gut diseases to tolerate a wider range of foods.

[0148] The dietary fibre formulation of the invention may also be used in a method or use for assisting those persons with gestational diabetes mellitus (GDM), and those with disabilities, neurological or mental health conditions, and those with other chronic disease where medication use negatively impacts the microbiome of the gut.

[0149] Each document, reference, patent application or patent cited in this text is expressly incorporated herein in their entirety by reference, which means that it should be read and considered by the reader as part of this text. That the document, reference, patent application, or patent cited in this text is not repeated in this text is merely for reasons for conciseness. Inclusion does not constitute an admission is made that any of the references constitute priorart or are part of the common general knowledge of those working in the field to which this invention relates.

[0150] Optional embodiments of the present invention may also be said to broadly consist in the parts, elements and features referred to or indicated herein, individually or collectively, in any or all combinations of two or more of the parts, elements or features, and wherein specific integers are mentioned herein which have known equivalents in the art to which the invention relates, such known equivalents are deemed to be incorporated herein as if individually set forth.

[0151] It is to be appreciated that reference to "one example" or "an example" of the invention is not made in an exclusive sense. Accordingly, one example may exemplify certain aspects of the invention, whilst other aspects are exemplified in a different example. These examples are intended to assist the skilled person in performing the invention and are not intended to limit the overall scope of the invention in any way unless the context clearly indicates otherwise.

[0152] It is to be understood that the terminology employed above is for the purpose of description and should not be regarded as limiting. The described embodiment is intended to be illustrative of the invention, without limiting the scope thereof. The invention is capable of being practised with various modifications and additions as will readily occur to those skilled in the art.

[0153] Other definitions for selected terms used herein may be found within the detailed description of the invention and apply throughout. Unless otherwise defined, all other scientific and technical terms used herein have the same meaning as commonly understood to one of ordinary skill in the art to which the invention belongs.

[0154] Various substantially and specifically practical and useful exemplary embodiments of the claimed person matter are described herein, textually and / or graphically, including the best mode, if any, known to the inventors for carrying out the claimed person matter.

[0155] Those skilled in the art will appreciate that the invention described herein is susceptible to variations and modifications other than those specifically described. It is to be understood that the invention includes all such variations and modifications. The invention also includes all of the steps, features, compositions and compounds referred to or indicatedin the specification, individually or collectively and any and all combinations or any two or more of the steps or features.

[0156] The inventor(s) expects skilled artisans to employ such variations as appropriate, and the inventor(s) intends for the claimed person matter to be practiced other than as specifically described herein. Accordingly, as permitted by law, the claimed person matter includes and covers all equivalents of the claimed person matter and all improvements to the claimed person matter. Moreover, every combination of the above described elements, activities, and all possible variations thereof are encompassed by the claimed person matter unless otherwise clearly indicated herein, clearly and specifically disclaimed, or otherwise clearly contradicted by context.

[0157] The present invention is not to be limited in scope by the specific embodiments described herein, which are intended for the purpose of exemplification only. Functionally equivalent products, compositions and methods are clearly within the scope of the invention as described herein.

[0158] The use of any and all examples, or exemplary language (e.g., "such as" or “for example”) provided herein, is intended merely to better illuminate one or more embodiments and does not pose a limitation on the scope of any claimed person matter unless otherwise stated. No language in the specification should be construed as indicating any non-claimed person matter as essential to the practice of the claimed person matter.

[0159] Throughout the specification and claims, unless the context requires otherwise, the word “comprise” or variations such as “comprises” or “comprising”, will be understood to imply the inclusion of a stated integer or group of integers but not the exclusion of any other integer or group of integers.

[0160] Throughout the specification unless the context requires otherwise, the word “include” or variations such as “includes” or “including”, will be understood to imply the inclusion of a stated integer or group of integers but not the exclusion of any other integer or group of integers.

[0161] Moreover, when any number or range is described herein, unless clearly stated otherwise, that number or range is approximate. Recitation of ranges of values herein are merely intended to serve as a shorthand method of referring individually to each separate value falling within the range, unless otherwise indicated herein, and each separate valueand each separate sub-range defined by such separate values is incorporated into the specification as if it were individually recited herein. For example, if a range of 1 to 10 is described, that range includes all values there between, such as for example, 1.1 , 2.5, 3.335, 5, 6.179, 8.9999, etc., and includes all sub-ranges there between, such as for example, 1 to 3.65, 2.8 to 8.14, 1.93 to 9, etc.

[0162] Accordingly, every portion (e.g., title, field, background, summary, description, abstract, drawing figure, etc.) of this application, other than the claims themselves, is to be regarded as illustrative in nature, and not as restrictive; and the scope of person matter protected by any patent that issues based on this application is defined only by the claims of that patent.

[0163] While there are shown and described presently further embodiments of the application, it is to be distinctly understood that the application is not limited thereto but may be otherwise variously embodied and practiced within the scope of the following claims.

[0164] Any of the features of an embodiment of aspects is applicable to all other aspects and embodiments identified herein. Any of the features of an embodiment is independently combinable, partly or wholly with other embodiments described herein in any way, e.g., one, two, or three or more embodiments may be combinable in whole or in part.ReferencesSo, D., Yao, C. K., Ardalan, Z. S., Thwaites, P. A., Kalantar-Zadeh, K., Gibson, P. R., & Muir, J. G. (2022). Supplementing dietary fibers with a low fodmap diet in irritable bowel syndrome: A randomized controlled crossover trial. Clinical Gastroenterology and Hepatology, 20(9), 2112-212O.e2117. https: / / doi.Org / 10.1016 / j .eg h .2021 .12.016Mysonhimer, A. R., & Holscher, H. D. (2022). Gastrointestinal effects and tolerance of non- digestible carbohydrate consumption. Advances in Nutrition, nmac094. https: / / doi.org / 10.1093 / advances / nmac094Drossman DA, Hasler WL. Rome IV-Functional Gl Disorders: Disorders of Gut-Brain Interaction. Gastroenterology. 2016;150(6):1257-61. https: / / doi.Org / 10.1053 / j.gastro.2016.03.035Zarrin R, I biebele Tl, Marks GC. Development and validity assessment of a diet quality index for Australians. Asia Pacific Journal of Clinical Nutrition. 2013;22(2):177- 87. https: / / doi.Org / 10.6133 / apjcn.2013.22.2.15Hlaing-Hlaing H, Pezdirc K, Tavener M, James EL, Hure A. Diet quality indices used in Australian and New Zealand adults: A systematic review and critical appraisal. Nutrients. 2020;12(12). https: / / doi.Org / 10.3390 / nu12123777National Health and Medical Research Council. Eat for Health: Australian Dietary Guidelines. In: Council NHaMR, editor. Canberra, Australia2013Nutrient Reference Values for Australia and New Zealand (nrv.gov.au) [Internet]. 2017 [cited 24 / 01 / 2022], Available from: https: / / www.nrv.gov.au / chronic-disease / summary.Stinson LF, Boyce MC, Payne MS, Keelan JA. The not-so-sterile womb: Evidence that the human fetus is exposed to bacteria prior to birth. Frontiers in Microbiology. 2019;10:1124. https : / / doi .org / 10.3389 / fmicb.2019.01124Jones J, Reinke SN, Ali A, Palmer DJ, Christophersen CT. Fecal sample collection methods and time of day impact microbiome composition and short chain fatty acid concentrations. Scientific Reports. 2021 ;11 (1 ):13964. https : / / doi .org / 10.1038 / s41598-021 -93031 -zMartin M. Cutadapt removes adapter sequences from high-throughput sequencing reads. EMBnetjournal. 2011 ;17(1):10-2.https: / / doi.org / 10.14806 / ej.17.1 .200Callahan BJ, McMurdie PJ, Rosen MJ, Han AW, Johnson AJA, Holmes SP. DADA2: High- resolution sample inference from Illumina amplicon data. Nature Methods. 2016; 13(7) :581 - 3. https: / / doi.org / 10.1038 / nmeth.3869Parks DH, Chuvochina M, Waite DW, Rinke C, Skarshewski A, Chaumeil P-A, et al. A standardized bacterial taxonomy based on genome phylogeny substantially revises the tree of life. Nature Biotechnology. 2018;36(10):996-1004. https: / / doi.org / 10.1038 / nbt.4229Birkett AM, Jones GP, deSilva AM, Young GP, Muir JG. Dietary intake and faecal excretion of carbohydrate by Australians: importance of achieving stool weights greater than 150 g to improve faecal markers relevant to colon cancer risk. European Journal of Clinical Nutrition. 1997;51 (9):625-32.https: / / doi. org / 10.1038 / sj.ejcn.1600456IBM. IBM SPSS Statistics for Windows. 25.0 ed. Armonk, NY: IBM Corp.; 2017Yan R, Murphy M, Genoni A, Marlow E, Dunican IC, Lo J, et al. Does Fibre-fix provided to people with irritable bowel syndrome who are consuming a low FODMAP diet improve their gut health, gut microbiome, sleep and mental health? A double-blinded, randomised controlled trial. BMJ Open Gastroenterology.2020;7(1 ):e000448. https: / / doi.Org / 10.1136 / bmjgast-2020-000448Faul F, Erdfelder E, Lang A-G, Buchner A. G*Power 3: A flexible statistical power analysis program for the social, behavioral, and biomedical sciences. Behavior Research Methods. 2007;39(2):175-91 .https: / / doi.Org / 10.3758 / BF03193146Zarrin R, I biebele Tl, Marks GC. Development and validity assessment of a diet quality index for Australians. Asia Pacific Journal of Clinical Nutrition. 2013;22(2):177- 87. https: / / d0i.0rg / l 0.6133 / apjcn.2013.22.2.15Dunican IC, Murray K, Slater JA, Maddison KJ, Jones MJ, Dawson B, et al. Laboratory and home comparison of wrist-activity monitors and polysomnography in middle-aged adults.Sleep and Biological Rhythms. 2017;16(1 ):85-97. https: / / doi.org / 10.1007 / s41105-017-0130- xRees J, Radavelli Bagatini S, Lo J, Hodgson JM, Christophersen CT, Daly RM, et al. 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Claims

The Claims Defining the Invention are as Follows:

1. A method of treating a person suffering from Irritable Bowel Syndrome (IBS), the method comprising: administering to the person a dietary fibre formulation comprising a ratio of approximately 3 parts high amylose maize starch (HAMS) to 1 part psyllium; wherein the person is administered increasing volumes of the formulation for at least two weeks in an initial ramp-up period; wherein the psyllium protects against fermentation of the HAMS as the formulation passes through the proximal colon, and wherein the HAMS can be fermented in the distal colon of the person to produce butyrate and faeces; and wherein the symptoms of IBS in the person are not substantially aggravated by administration of the formulation.

2. A method according to claim 1 , wherein the initial ramp up period comprises approximately three weeks.

3. A method according to claim 1 or claim 2, wherein the initial ramp up period comprises two days of 5 g dose of dietary fibre formulation.

4. A method according to any one of the preceding claims, wherein the initial ramp up period comprises two days of 5 g / day dose of dietary fibre formulation followed by two days of 10 g / day dose of dietary fibre formulation.

5. A method according to any one of the preceding claims, wherein the initial ramp up period comprises two days of 5 g / day dose of dietary fibre formulation followed by two days of 10 g / day dose of dietary fibre formulation, followed by three days of 20 g / day dose of dietary fibre formulation.

6. A method according to any one of the preceding claims, wherein the initial ramp up period comprises two days of 5 g / day dose of dietary fibre formulation followed by two days of 10 g / day dose of dietary fibre formulation, followed by three days of 20 g / day dose of dietary fibre formulation, followed by 11 days of 40 g / day dose of dietary fibre formulation.

7. A method according to any one of the preceding claims, wherein the ramp up period of increasing amounts of dietary fibre formulation enables the gut biome of the person to adjust to the increase in total dietary fibre and fermentable fibres.

8. A method according to any one of the preceding claims, wherein following the initial ramp up period is a period of maintenance comprising a dose of between approximately 20 g / day to 30 g / day dose of dietary fibre formulation.

9. A method according to any one of claims 3 to 8, wherein the dose is divided into more than one administration per day.

10. A method according to claim 9, wherein half of the dose is administered twice a day, or a third of the dose is administered three times a day.

11. A method according to any one of the preceding claims, wherein the formulation can be administered as a dietary supplement.

12. A method according to any one of claims 1 to 10, wherein the formulation can be administered in combination with one or more food products.

13. A method according to any one of the preceding claims, wherein the persons comprise athletes or sportspeople.

14. A method according to any one of claims 1 to 12, wherein the persons comprise those taking the formulation for weight loss.

15. A method according to any one of claims 1 to 12, wherein the persons comprise those taking the formulation as a preventative for colorectal cancer due to increasing production of butyrate in the colon of the person.

16. A method according to any one of claims 1 to 12, wherein the persons comprise those taking the formulation for gut repair and where medication use negatively effects the persons microbiome in their gut.