Anaerostipes sp. and bacteroides uniformis prebiotic composition and method of use

Abstract

A prebiotic fiber composition obtained from Gloeoporus thelephoroides, Ganoderma lucidum, Pholiota nameko, and / or Pleurotus pulmonarius; and a method of promoting growth of Anaerostipes sp., Bacteroides uniformis, or both in the gut microbiome of an animal, which method comprises orally administering the prebiotic fiber composition, alone or in further combination with live Anaerostipes sp., Bacteroides uniformis, or both, simultaneously or sequentially.

Description

70085-02 ANAEROSTIPES SP. AND BACTEROIDES UNIFORMIS PREBIOTIC COMPOSITION AND METHOD OF USE CROSS-REFERENCE TO RELATED APPLICATION

[0001] This application claims priority to U.S. provisional patent application no. 63 / 535,387, which was filed August 30, 2023, and the content of which is hereby incorporated by reference in its entirety. TECHNICAL FIELD

[0002] The present disclosure is directed to a prebiotic fiber composition that supports the beneficial gut bacteria Anaerostipes sp. and Bacteroides uniformis and a method of use. SEQUENCE LISTING

[0003] A computer-readable form (CRF) of the Sequence Listing is submitted with this application. The sequence listing is entitled 70085-02_SEQ_LISTING.xml, was generated on August 20, 2024, and is 13000 bytes in size. The entire content of the sequence listing is incorporated herein by reference in its entirety. BACKGROUND

[0004] Prebiotics are carbohydrate-based dietary fibers that support beneficial bacteria in the gut. A problem with prebiotics, however, is that commercially available products do not target specific beneficial gut bacteria in a consistent way. Consequently, the outcome of prebiotic ingestion differs from one person to the next. Overall, the efficacy of commercially available products has not been established. Further, current prebiotics do not support next- generation probiotics.

[0005] The present disclosure seeks to address a long-felt and unmet need by matching prebiotics with beneficial gut bacteria. This and other objects and advantages, as well as inventive features, will be apparent from the detailed description provided herein.70085-02 SUMMARY

[0006] A prebiotic fiber composition obtained from Gloeoporus thelephoroides (GT) is provided. The prebiotic fiber composition can be obtained by any suitable method. An example of such a method comprises: (i) grinding, or obtaining ground, lyophilized fruiting bodies of GT; (ii) dissolving the ground, lyophilized fruiting bodies of GT in distilled water, or in lieu of steps (i) and (ii), obtaining ground, lyophilized fruiting bodies of GT dissolved in distilled water and optionally reduced with NaBH4; (iii) if not already reduced, reducing the ground, lyophilized fruiting bodies of GT dissolved in distilled water with NaBH4; (iv) subjecting the reduced, ground, lyophilized fruiting bodies of GT dissolved in distilled water to exhaustive hot-water extraction; (v) filtering the extraction to obtain a filtrate; (vi) subjecting the filtrate of (v) to extraction in an alkaline medium; (vii) centrifuging the extraction of (vi) to obtain a supernatant; (viii) neutralizing, dialyzing and precipitating the supernatant of (vii); (ix) centrifuging the precipitated supernatant of (viii) to obtain a precipitate; and (x) lyophilizing the precipitate of (ix) or solubilizing the precipitate of (ix) in water, whereupon the prebiotic fiber composition from GT is obtained. The prebiotic fiber composition comprises (1→3),(1→6) and (1→3)-β-D-glucan. The exhaustive hot- water extraction can be carried out for about 3 hours (e.g., 3 hours) at about 100°C (e.g., 100°C) using a water bath under reflux. The alkaline medium of (vi) can comprise 5% potassium hydroxide. The centrifuging in (vii) and (ix) can be carried out at about 10,000 rpm (e.g., 10,000 rpm) at about 25 °C (e.g., 25 °C) for about 20 minutes (e.g., 20 minutes). The precipitating in (viii) can be carried out in ethanol (3:1 v / v).

[0007] In view of the above, also provided is a prebiotic fiber composition obtained from GT. The prebiotic fiber composition comprises (1→3),(1→6) and (1→3)- β-D-glucan. The prebiotic fiber composition can be lyophilized. The prebiotic fiber composition can be insoluble or solubilized in water. The prebiotic fiber composition can be more effective in its insoluble form.70085-02

[0008] Also in view of the above, a method of promoting growth of Anaerostipes sp., Bacteroides uniformis, or both in the gut microbiome of an animal (e.g., warm-blooded animal, e.g., mammal, e.g., human) is provided. The method comprises orally administering an above- described prebiotic fiber composition obtained from GT, alone or in further combination with live Anaerostipes sp., Bacteroides uniformis, or both, simultaneously (e.g., as one, two or three compositions) or sequentially (e.g., as two or three compositions), whereupon growth of Anaerostipes sp., Bacteroides uniformis, or both in the gut microbiome of the animal is promoted.

[0009] A prebiotic fiber composition obtained from Ganoderma lucidum (GL) is further provided. The prebiotic fiber composition can be obtained by any suitable method, such as a method comprising: (i) grinding, or obtaining ground, lyophilized fruiting bodies of GL; (ii) dissolving the ground, lyophilized fruiting bodies of GL in distilled water, or in lieu of steps (i) and (ii), obtaining ground, lyophilized fruiting bodies of GL dissolved in distilled water and optionally reduced with NaBH4; (iii) if not already reduced, reducing the ground, lyophilized fruiting bodies of GL dissolved in distilled water with NaBH4; (iv) subjecting the reduced, ground, lyophilized fruiting bodies of GL dissolved in distilled water to exhaustive hot-water extraction; (v) filtering the extraction to obtain a filtrate; (vi) subjecting the filtrate of (v) to extraction in an alkaline medium; (vii) centrifuging the extraction of (vi) to obtain a supernatant; (viii) neutralizing, dialyzing and precipitating the supernatant of (vii); (ix) centrifuging the precipitated supernatant of (viii) to obtain a precipitate; and (x) lyophilizing the precipitate of (ix) or solubilizing the precipitate of (ix) in water, whereupon the prebiotic fiber composition from GL is obtained. The prebiotic fiber composition comprises (1→3),(1→6) and (1→3)-β-D-glucan. The exhaustive hot-water extraction can be carried out for about 3 hours (e.g., 3 hours) at about 100°C (e.g., 100°C) using a water bath under reflux. The alkaline medium of (vi) can comprise 5% potassium hydroxide. The centrifuging in (vii) and (ix) can be carried out at about 10,000 rpm (e.g., 10,000 rpm) at about70085-02 25 °C (e.g., 25 °C) for about 20 minutes (e.g., 20 minutes). The precipitating in (viii) can be carried out in ethanol (3:1 v / v).

[0010] In view of the above, also provided is a prebiotic fiber composition obtained from GL. The prebiotic fiber composition comprises (1→3),(1→6) and (1→3)- β-D-glucan. The prebiotic fiber composition can be lyophilized. The prebiotic fiber composition can be insoluble or solubilized in water. The prebiotic fiber composition can be more effective in its insoluble form.

[0011] Also in view of the above, a method of promoting growth of Anaerostipes sp., Bacteroides uniformis, or both in the gut microbiome of an animal (e.g., warm- blooded animal, e.g., mammal, e.g., human) is provided. The method comprises orally administering an above-described prebiotic fiber composition obtained from GL, alone or in further combination with live Anaerostipes sp., Bacteroides uniformis, or both, simultaneously (e.g., as one, two or three compositions) or sequentially (e.g., as two or three compositions), whereupon growth of Anaerostipes sp., Bacteroides uniformis, or both in the gut microbiome of the animal is promoted.

[0012] A prebiotic fiber composition obtained from Pholiota nameko (PN) is still further provided. The prebiotic fiber composition can be obtained by any suitable method, such as a method comprising: (i) grinding, or obtaining ground, lyophilized fruiting bodies of PN; (ii) dissolving the ground, lyophilized fruiting bodies of PN in distilled water, or in lieu of steps (i) and (ii), obtaining ground, lyophilized fruiting bodies of PN dissolved in distilled water and optionally reduced with NaBH4; (iii) if not already reduced, reducing the ground, lyophilized fruiting bodies of PN dissolved in distilled water with NaBH4; (iv) subjecting the reduced, ground, lyophilized fruiting bodies of PN dissolved in distilled water to extraction under high temperature and high pressure to obtain an extract; (v) filtering the extract of (iv) to obtain an eluant; (vi) precipitating the eluant from (v) to obtain a precipitated supernatant; (vii) centrifuging the precipitated supernatant of (vi); (viii) solubilizing the precipitate of (vii) in water;70085-02 (ix) freeze-thawing the solubilized precipitate of (viii) three times to obtain soluble and insoluble fractions; (x) dissolving the soluble fraction of (ix) in water; (xi) ultra-filtering the dissolved soluble fraction of (x) using a membrane with a 3kDa cut-off, and (xii) lyophilizing the filtrate of (ix) or solubilizing the filtrate of (ix) in water,

[0013] whereupon the prebiotic fiber composition from PN is obtained. The prebiotic fiber composition comprises (1→3),(1→6) and (1→3)-β-D-glucan. The extraction under high temperature and high pressure can be carried out for about one hour (one hour) at about 121 °C (e.g., 121 °C) and about 1.2 atm (e.g., 1.2 atm) using an autoclave. The precipitating in (vi) can be carried out in ethanol (3:1 v / v). The centrifuging in (vii) can be carried out at about 10,000 rpm (e.g., 10,000 rpm) at about 25 °C (e.g., 25 °C) for about 20 minutes (e.g., 20 minutes).

[0014] In view of the above, also provided is a prebiotic fiber composition obtained from PN. The prebiotic fiber composition comprises (1→3),(1→6) and (1→3)-β-D-glucan. The prebiotic fiber composition can be lyophilized. The prebiotic fiber composition can be insoluble or solubilized in water. The prebiotic fiber composition can be more effective in its insoluble form.

[0015] Also in view of the above, a method of promoting growth of Anaerostipes sp., Bacteroides uniformis, or both in the gut microbiome of an animal (e.g., warm-blooded animal, e.g., mammal, e.g., human) is provided. The method comprises orally administering the prebiotic fiber composition obtained from PN, alone or in further combination with live Anaerostipes sp., Bacteroides uniformis, or both, simultaneously (e.g., as one, two or three compositions) or sequentially (e.g., as two or three compositions), whereupon growth of Anaerostipes sp., Bacteroides uniformis, or both in the gut microbiome of the animal is promoted.

[0016] A prebiotic fiber composition obtained from Pleurotus pulmonarius (PP) is even still further provided. The prebiotic fiber composition can be obtained by any suitable method, such as a method comprising: (i) grinding, or obtaining ground, lyophilized fruiting bodies of PP;70085-02 (ii) dissolving the ground, lyophilized fruiting bodies of PP in distilled water, or in lieu of steps (i) and (ii), obtaining ground, lyophilized fruiting bodies of PP dissolved in distilled water and optionally reduced with NaBH4; (iii) if not already reduced, reducing the ground, lyophilized fruiting bodies of PP dissolved in distilled water with NaBH4; (iv) subjecting the reduced, ground, lyophilized fruiting bodies of PN dissolved in distilled water to extraction under high temperature and high pressure; (v) filtering the extraction to obtain a filtrate; (vi) extracting the filtrate of (v) in an alkaline medium to obtain an extraction; (vii) centrifuging the extraction of (vi) to obtain a supernatant; (viii) neutralizing, dialyzing and precipitating the supernatant; (ix) centrifuging the precipitated supernatant of (viii) to obtain a precipitate; (x) solubilizing the precipitate of (ix) in water; (xi) freeze-thawing the solubilized precipitate of (x) three times to obtain soluble and insoluble fractions; and (xii) lyophilizing the precipitate of (ix) or solubilizing the precipitate of (ix) in water, whereupon the prebiotic fiber composition from PP is obtained. The prebiotic composition comprises (1→3),(1→6) and (1→3)-β-D-glucan. The extraction under high temperature and high pressure can be carried out for about one hour (e.g., one hour) at about 121 °C (e.g., 121 °C) and about 1.2 atm (e.g., 1.2 atm) using an autoclave. The alkaline medium of (vi) can comprise 5% potassium hydroxide. The centrifuging in (vii) and (ix) can be carried out at about 10,000 rpm (e.g., 10,000 rpm) at about 25 °C (e.g., 25 °C) for about 20 minutes (e.g., 20 minutes). The precipitating in (viii) can be carried out in ethanol (3:1 v / v).

[0017] In view of the above, also provided is a prebiotic fiber composition obtained from PP. The prebiotic fiber composition comprises (1→3),(1→6) and (1→3)-70085-02 β-D-glucan. The prebiotic fiber composition can be lyophilized. The prebiotic fiber composition can be insoluble or solubilized in water. The prebiotic fiber composition can be more effective in its insoluble form.

[0018] Also in view of the above, a method of promoting growth of Anaerostipes sp., Bacteroides uniformis, or both in the gut microbiome of an animal (e.g., warm-blooded animal, e.g., mammal, e.g., human) is provided. The method comprises orally administering the prebiotic fiber composition obtained from PP, alone or in further combination with live Anaerostipes sp., Bacteroides uniformis, or both, simultaneously (e.g., as one or more compositions) or sequentially, whereupon growth of Anaerostipes sp., Bacteroides uniformis, or both in the gut microbiome of the animal is promoted.

[0019] Provided also is a prebiotic fiber composition comprising: (a) a prebiotic fiber composition obtained from Gloeoporus thelephoroides (GT), which comprises (1→3),(1→6) and (1→3)-β-D-glucan; (b) a prebiotic fiber composition obtained from Ganoderma lucidum (GL), which comprises (1→3),(1→6) and (1→3)-β-D-glucan; (c) a prebiotic fiber composition obtained from Pholiota nameko (PN), which comprises (1→3),(1→6) and (1→3)-β-D-glucan; (d) a prebiotic fiber composition obtained from Pleurotus pulmonarius (PP), which comprises (1→3),(1→6) and (1→3)-β-D-glucan; (e) (a) and (b); (f) (a) and (c); (g) (a) and (d); (h) (b) and (c); (i) (b) and (d); (j) (c) and (d); or (k) (a), (b), (c), and (d). The prebiotic fiber composition can be lyophilized. The prebiotic fiber composition can be insoluble or solubilized in water. The prebiotic fiber composition can be more effective in its insoluble form.

[0020] In view of the prebiotic fiber composition described immediately above, also provided is a method of promoting growth of Anaerostipes sp., Bacteroides uniformis, or both in the gut microbiome of an animal (e.g., warm-blooded animal, e.g., mammal, e.g., human). The70085-02 method comprises orally administering the prebiotic fiber composition described immediately above, alone or in further combination with live Anaerostipes sp., Bacteroides uniformis, or both, simultaneously (e.g., as one or more compositions) or sequentially (e.g., as one or more compositions), whereupon growth of Anaerostipes sp., Bacteroides uniformis, or both in the gut microbiome of the animal is promoted. FIGURES

[0021] Fig.1. Short chain fatty acid production (mM) of 24 h in vitro fecal fermentation of fractions PP, PPSm, PN and PNSm compared to fructoologiosaacgarides (FOS) and the blanks at 0 h and 24 h. (A) Total SCFAs (B) Acetate, (C) Propionate, (D) Butyrate. Statistical analysis of pairwise comparisons is presented in Supplementary Table 1. (E) Proportion of each SCFA related to the total amount of SCFAs produced (%).

[0022] Fig.2. Relative abundance of selected bacterial species from in vitro fecal fermented fractions PP, PPSm, PN and PNSm compared to fructoologiosaacgarides (FOS) and the blanks at 0 h and 24 h. (A) Anaerostipes sp., (B) Bacteroides uniformis, (C) Bacteroides ovatus, (D) Roseburia faecis, and (E) Parabacteroides distasonis.

[0023] Fig.3. Anaerostipes sp. is an unclassified species in the Anaerostipes genus. Depending on the database used, it can be classified in a different genus. For example, Anaerostipes sp was classified in the Anaerostipes genus when Greengenes 13_8 was used but, when Silva 123 was used for classification, the same sequences were classified as belonging to the Eubacterium ventriosium group. In order to avoid any potential ambiguity, the sequences of the amplicon sequence variants (ASVs) are set forth here.

[0024] Fig.4. Relative abundance of Anaerostipes sp. from in vitro fecal fermented fractions PP, PPSm, PN, and PNSm compared to fructooligosaccharides (FOS) and blanks at 0 hour and 24 hours. The insoluble branched β-D-Glucan PP is highly effective in promoting Anaerostipes sp., with superiority compared to the soluble branched β-D-Glucan PN, which also promoted Anaerostipes sp. to some extent. Importantly, debranching of PP and PN (fractions PPSm and PNSm, respectively) completely prevents Anaerostipes promotion, indicating the importance of details in the70085-02 physico-chemical structure for Anaerostipes sp. promotion. PP = branched, insoluble β-D- glucan; PN = branched, soluble β-D-glucan; PPSm = debranched β-D-glucan PP; and PNSm debranched β-D-glucan PN. DETAILED DESCRIPTION

[0025] The most common dietary fiber present in mushrooms are the branched (1→3),(1→6)-linked β-D-glucans (Ruthes et al., 2013; Santos-Neves et al., 2008; Smiderle et al., 2006). In general, they consist mainly of a backbone of β-D-Gclp (1→3)-linked units partially substituted at O-6 by side chains of β-D-Glcp (1→3) or (1→6)-linked units and / or single β-D- Glcp residues as non-reducing end units (Bhanja et al., 2014; Moradali et al., 2007; Moreno et al., 2016; Zhu et al., 2015). Such β-glucans differ from those found in cereals, which have (1→3) and (1→4), but not (1→6), linkages and branching structures. The distinct set of polysaccharides’ physico-chemical structures found in mushrooms offers the possibility of a variety of unique biological activities (Cerletti et al., 2021).

[0026] Since the human body does not produce enzymes to digest (1→3),(1→6)-linked β-D-glucans, they arrive intact in the large intestine, where they can directly act on immune cells, and are fermented by resident gut bacteria, which produce short-chain fatty acids (SCFAs) and promote bacterial shifts beneficial to the host (Ruthes et al., 2021; van Steenwijk et al., 2021). Cantu-Jungles et al. (2018) previously showed that insoluble (1→3),(1→6) and (1→3) β-glucans isolated from the mushroom Cookeina speciosa can modulate the human gut microbiota in a targeted way with promotion of health-related bacteria such as Anaerostipes sp. and Bacteroides uniformis, even if tested in the gut microbiota of different individuals (Cantu-Jungles et al., 2021). Interestingly, several other studies using other mushroom (1→3),(1→6)-linked β-D- glucans did not show specific promotion of these bacteria during in vitro fecal fermentations, thus highlighting the importance of discrete features in glucans that can change gut microbiota outcomes, as previously suggested (Ruthes et al., 2021). Since Cookeina speciosa mushroom is not commercially available, let alone distributed worldwide, (1→3),(1→6)-linked β-D-glucans from other sources (Gloeoporus thelephoroides, Ganoderma lucidum, Pleurotus nameko and Pleurotus pulmonarius) were evaluated for targeted promotion of Anaerostipes sp. and B. uniformis in the human gut microbiota. Anaerostipes sp. and Bacteroides uniformis reduce local and systemic inflammation related to various chronic health conditions and diseases. The β-70085-02 glucans (1-3, 1-6) found in Gannoderma lucidum (GL; commercially known as Reishi mushroom), Pleurotus pulmonarius (PP; commercially known as Italian oyster mushroom), Pholiota microspora (PM), and Gloeoporus thelephoroides (GT) promote Anaerostipes sp. and Bacteroides uniformis better than fructooligosaccharides. Given the ability of Anaerostipes sp. and Bacteroides uniformis to reduce inflammation, they have been identified as potential next-generation probiotic bacteria. The β-glucans provided herein support the bacteria and their growth. The β-glucans can be used as a prebiotic or as a synbiotic to promote a probiotic.

[0027] In view of the above, provided is a prebiotic fiber composition obtained from Gloeoporus thelephoroides (GT). The prebiotic fiber composition can be obtained by any suitable method, such as a method comprising: (i) grinding, or obtaining ground, lyophilized fruiting bodies of GT; (ii) dissolving the ground, lyophilized fruiting bodies of GT in distilled water, or in lieu of steps (i) and (ii), obtaining ground, lyophilized fruiting bodies of GT dissolved in distilled water and optionally reduced with NaBH4; (iii) if not already reduced, reducing the ground, lyophilized fruiting bodies of GT dissolved in distilled water with NaBH4; (iv) subjecting the reduced, ground, lyophilized fruiting bodies of GT dissolved in distilled water to exhaustive hot-water extraction; (v) filtering the extraction to obtain a filtrate; (vi) subjecting the filtrate of (v) to extraction in an alkaline medium; (vii) centrifuging the extraction of (vi) to obtain a supernatant; (viii) neutralizing, dialyzing and precipitating the supernatant of (vii); (ix) centrifuging the precipitated supernatant of (viii) to obtain a precipitate; and (x) lyophilizing the precipitate of (ix) or solubilizing the precipitate of (ix) in water, whereupon the prebiotic fiber composition from GT is obtained. The prebiotic fiber composition comprises (1→3),(1→6) and (1→3)-β-D-glucan. The exhaustive hot- water extraction can be carried out for about 3 hours (e.g., 3 hours) at about 100°C (e.g., 100°C) using a water bath under reflux. The alkaline medium of (vi) can comprise 5% potassium hydroxide. The centrifuging in (vii) and (ix) can be carried out at about70085-02 10,000 rpm (e.g., 10,000 rpm) at about 25 °C (e.g., 25 °C) for about 20 minutes (e.g., 20 minutes). The precipitating in (viii) can be carried out in ethanol (3:1 v / v).

[0028] In view of the above, also provided is a prebiotic fiber composition obtained from GT. The prebiotic fiber composition comprises (1→3),(1→6) and (1→3)-β-D-glucan. The prebiotic fiber composition can be lyophilized. The prebiotic fiber composition can be insoluble or solubilized in water. The prebiotic fiber composition can be more effective in its insoluble form.

[0029] Also in view of the above, a method of promoting growth of Anaerostipes sp., Bacteroides uniformis, or both in the gut microbiome of an animal (e.g., warm-blooded animal, e.g., mammal, e.g., human) is provided. The method comprises orally administering an above- described prebiotic fiber composition obtained from GT, alone or in further combination with live Anaerostipes sp., Bacteroides uniformis, or both, simultaneously (e.g., as one, two or three compositions) or sequentially (e.g., as two or three compositions), whereupon growth of Anaerostipes sp., Bacteroides uniformis, or both in the gut microbiome of the animal is promoted.

[0030] A prebiotic fiber composition obtained from Ganoderma lucidum (GL) is further provided. The prebiotic fiber composition can be obtained by any suitable method, such as a method comprising: (i) grinding, or obtaining ground, lyophilized fruiting bodies of GL; (ii) dissolving the ground, lyophilized fruiting bodies of GL in distilled water, or in lieu of steps (i) and (ii), obtaining ground, lyophilized fruiting bodies of GL dissolved in distilled water and optionally reduced with NaBH4; (iii) if not already reduced, reducing the ground, lyophilized fruiting bodies of GL dissolved in distilled water with NaBH4; (iv) subjecting the reduced, ground, lyophilized fruiting bodies of GL dissolved in distilled water to exhaustive hot-water extraction; (v) filtering the extraction to obtain a filtrate; (vi) subjecting the filtrate of (v) to extraction in an alkaline medium; (vii) centrifuging the extraction of (vi) to obtain a supernatant; (viii) neutralizing, dialyzing and precipitating the supernatant of (vii); (ix) centrifuging the precipitated supernatant of (viii) to obtain a precipitate; and70085-02 (x) lyophilizing the precipitate of (ix) or solubilizing the precipitate of (ix) in water, whereupon the prebiotic fiber composition from GL is obtained. The prebiotic fiber composition comprises (1→3),(1→6) and (1→3)-β-D-glucan. The exhaustive hot-water extraction can be carried out for about 3 hours (e.g., 3 hours) at about 100°C (e.g., 100°C) using a water bath under reflux. The alkaline medium of (vi) can comprise 5% potassium hydroxide. The centrifuging in (vii) and (ix) can be carried out at about 10,000 rpm (e.g., 10,000 rpm) at about 25 °C (e.g., 25 °C) for about 20 minutes (e.g., 20 minutes). The precipitating in (viii) can be carried out in ethanol (3:1 v / v).

[0031] In view of the above, also provided is a prebiotic fiber composition obtained from GL. The prebiotic fiber composition comprises (1→3),(1→6) and (1→3)- β-D-glucan. The prebiotic fiber composition can be lyophilized. The prebiotic fiber composition can be insoluble or solubilized in water. The prebiotic fiber composition can be more effective in its insoluble form.

[0032] Also in view of the above, a method of promoting growth of Anaerostipes sp., Bacteroides uniformis, or both in the gut microbiome of an animal (e.g., warm- blooded animal, e.g., mammal, e.g., human) is provided. The method comprises orally administering an above-described prebiotic fiber composition obtained from GL, alone or in further combination with live Anaerostipes sp., Bacteroides uniformis, or both, simultaneously (e.g., as one, two or three compositions) or sequentially (e.g., as two or three compositions), whereupon growth of Anaerostipes sp., Bacteroides uniformis, or both in the gut microbiome of the animal is promoted.

[0033] A prebiotic fiber composition obtained from Pholiota nameko (PN) is still further provided. The prebiotic fiber composition can be obtained by any suitable method, such as a method comprising: (i) grinding, or obtaining ground, lyophilized fruiting bodies of PN; (ii) dissolving the ground, lyophilized fruiting bodies of PN in distilled water, or in lieu of steps (i) and (ii), obtaining ground, lyophilized fruiting bodies of PN dissolved in distilled water and optionally reduced with NaBH4; (iii) if not already reduced, reducing the ground, lyophilized fruiting bodies of PN dissolved in distilled water with NaBH4;70085-02 (iv) subjecting the reduced, ground, lyophilized fruiting bodies of PN dissolved in distilled water to extraction under high temperature and high pressure to obtain an extract; (v) filtering the extract of (iv) to obtain an eluant; (vi) precipitating the eluant from (v) to obtain a precipitated supernatant; (vii) centrifuging the precipitated supernatant of (vi); (viii) solubilizing the precipitate of (vii) in water; (ix) freeze-thawing the solubilized precipitate of (viii) three times to obtain soluble and insoluble fractions; (x) dissolving the soluble fraction of (ix) in water; (xi) ultra-filtering the dissolved soluble fraction of (x) using a membrane with a 3kDa cut-off, and (xii) lyophilizing the filtrate of (ix) or solubilizing the filtrate of (ix) in water,

[0034] whereupon the prebiotic fiber composition from PN is obtained. The prebiotic fiber composition comprises (1→3),(1→6) and (1→3)-β-D-glucan. The extraction under high temperature and high pressure can be carried out for about one hour (one hour) at about 121 °C (e.g., 121 °C) and about 1.2 atm (e.g., 1.2 atm) using an autoclave. The precipitating in (vi) can be carried out in ethanol (3:1 v / v). The centrifuging in (vii) can be carried out at about 10,000 rpm (e.g., 10,000 rpm) at about 25 °C (e.g., 25 °C) for about 20 minutes (e.g., 20 minutes).

[0035] In view of the above, also provided is a prebiotic fiber composition obtained from PN. The prebiotic fiber composition comprises (1→3),(1→6) and (1→3)-β-D-glucan. The prebiotic fiber composition can be lyophilized. The prebiotic fiber composition can be insoluble or solubilized in water. The prebiotic fiber composition can be more effective in its insoluble form.

[0036] Also in view of the above, a method of promoting growth of Anaerostipes sp., Bacteroides uniformis, or both in the gut microbiome of an animal (e.g., warm-blooded animal, e.g., mammal, e.g., human) is provided. The method comprises orally administering the prebiotic fiber composition obtained from PN, alone or in further combination with live Anaerostipes sp., Bacteroides uniformis, or both, simultaneously (e.g., as one, two or three compositions) or sequentially (e.g., as two or three compositions), whereupon growth of Anaerostipes sp., Bacteroides uniformis, or both in the gut microbiome of the animal is promoted.70085-02

[0037] A prebiotic fiber composition obtained from Pleurotus pulmonarius (PP) is even still further provided. The prebiotic fiber composition can be obtained by any suitable method, such as a method comprising: (i) grinding, or obtaining ground, lyophilized fruiting bodies of PP; (ii) dissolving the ground, lyophilized fruiting bodies of PP in distilled water, or in lieu of steps (i) and (ii), obtaining ground, lyophilized fruiting bodies of PP dissolved in distilled water and optionally reduced with NaBH4; (iii) if not already reduced, reducing the ground, lyophilized fruiting bodies of PP dissolved in distilled water with NaBH4; (iv) subjecting the reduced, ground, lyophilized fruiting bodies of PN dissolved in distilled water to extraction under high temperature and high pressure; (v) filtering the extraction to obtain a filtrate; (vi) extracting the filtrate of (v) in an alkaline medium to obtain an extraction; (vii) centrifuging the extraction of (vi) to obtain a supernatant; (viii) neutralizing, dialyzing and precipitating the supernatant; (ix) centrifuging the precipitated supernatant of (viii) to obtain a precipitate; (x) solubilizing the precipitate of (ix) in water; (xi) freeze-thawing the solubilized precipitate of (x) three times to obtain soluble and insoluble fractions; and (xii) lyophilizing the precipitate of (ix) or solubilizing the precipitate of (ix) in water, whereupon the prebiotic fiber composition from PP is obtained. The prebiotic composition comprises (1→3),(1→6) and (1→3)-β-D-glucan. The extraction under high temperature and high pressure can be carried out for about one hour (e.g., one hour) at about 121 °C (e.g., 121 °C) and about 1.2 atm (e.g., 1.2 atm) using an autoclave. The alkaline medium of (vi) can comprise 5% potassium hydroxide. The centrifuging in (vii) and (ix) can be carried out at about 10,000 rpm (e.g., 10,000 rpm) at about 25 °C (e.g., 25 °C) for about 2070085-02 minutes (e.g., 20 minutes). The precipitating in (viii) can be carried out in ethanol (3:1 v / v).

[0038] In view of the above, also provided is a prebiotic fiber composition obtained from PP. The prebiotic fiber composition comprises (1→3),(1→6) and (1→3)-β-D-glucan. The prebiotic fiber composition can be lyophilized. The prebiotic fiber composition can be insoluble or solubilized in water. The prebiotic fiber composition can be more effective in its insoluble form.

[0039] Also in view of the above, a method of promoting growth of Anaerostipes sp., Bacteroides uniformis, or both in the gut microbiome of an animal (e.g., warm-blooded animal, e.g., mammal, e.g., human) is provided. The method comprises orally administering the prebiotic fiber composition obtained from PP, alone or in further combination with live Anaerostipes sp., Bacteroides uniformis, or both, simultaneously (e.g., as one or more compositions) or sequentially, whereupon growth of Anaerostipes sp., Bacteroides uniformis, or both in the gut microbiome of the animal is promoted.

[0040] Provided also is a prebiotic fiber composition comprising: (a) a prebiotic fiber composition obtained from Gloeoporus thelephoroides (GT), which comprises (1→3),(1→6) and (1→3)-β-D-glucan; (b) a prebiotic fiber composition obtained from Ganoderma lucidum (GL), which comprises (1→3),(1→6) and (1→3)-β-D-glucan; (c) a prebiotic fiber composition obtained from Pholiota nameko (PN), which comprises (1→3),(1→6) and (1→3)-β-D-glucan; (d) a prebiotic fiber composition obtained from Pleurotus pulmonarius (PP), which comprises (1→3),(1→6) and (1→3)-β-D-glucan; (e) (a) and (b); (f) (a) and (c); (g) (a) and (d); (h) (b) and (c); (i) (b) and (d); (j) (c) and (d); or (k) (a), (b), (c), and (d). The prebiotic fiber composition can be lyophilized. The prebiotic fiber composition can be solubilized in water.70085-02

[0041] In view of the prebiotic fiber composition described immediately above, also provided is a method of promoting growth of Anaerostipes sp., Bacteroides uniformis, or both in the gut microbiome of an animal (e.g., warm-blooded animal, e.g., mammal, e.g., human). The method comprises orally administering the prebiotic fiber composition described immediately above, alone or in further combination with live Anaerostipes sp., Bacteroides uniformis, or both, simultaneously (e.g., as one or more compositions) or sequentially (e.g., as one or more compositions), whereupon growth of Anaerostipes sp., Bacteroides uniformis, or both in the gut microbiome of the animal is promoted. EXAMPLES

[0042] The following examples serve to illustrate the present disclosure. The examples are not intended to limit the scope of the claimed invention in any way. Materials

[0043] Gloeoporus thelephoroides (GT) – The lyophilized fruiting bodies of the mushroom G. thelephoroides were ground and dissolved in distilled water (0.3 L / 10 g), reduced with a small aliquot of NaBH4and submitted to exhaustive hot-water extraction (3 hours, 100°C) using a water bath under reflux. The materials were filtered, and the retained fraction was subject to a new autoclave extraction, now in an alkaline medium with 5% potassium hydroxide, which was then centrifuged (20 minutes, 10,000 rpm, 25 °C). The supernatant was then neutralized, dialyzed and precipitated with ethanol (3:1, v / v), followed by centrifugation (20 minutes, 10,000 rpm, 25 °C). The precipitated fraction was solubilized in water, and characterized as a (1→3),(1→6) and (1→3)-β-D- glucan named GT (Table 1), which was later used for in vitro fecal fermentation experiments.

[0044] Ganoderma lucidum (GL) – The lyophilized fruiting bodies of the mushroom G. lucidum were ground and dissolved in distilled water (0.3 L / 10 g), reduced with a small aliquot of NaBH4and submitted to exhaustive hot-water extraction (3 hours, 100°C) using a water bath under reflux. The materials were filtered, and the retained fraction was subject to a new autoclave extraction, now in an alkaline medium with 5%70085-02 potassium hydroxide, which was then centrifuged (20 minutes, 10,000 rpm, 25 °C). The supernatant was then neutralized, dialyzed and precipitated with ethanol (3:1, v / v), followed by centrifugation (20 minutes, 10,000 rpm, 25 °C). The precipitate fraction was solubilized in water and characterized as a (1→3),(1→6) and (1→3)-β-D-glucan named GL (Table 1), which was later used for in vitro fecal fermentation experiments.

[0045] P. nameko glucan (PN) – The lyophilized fruiting bodies of the mushroom P. nameko were ground and dissolved in distilled water (0.3 L / 10 g), reduced with a small aliquot of NaBH4and submitted to high temperature and pressure extraction (121 °C, 1.2 atm, 1 hour) using an autoclave. The materials were filtered, and the eluted fraction was precipitated by addition of ethanol (3:1, v / v), followed by centrifugation (20 minutes, 10,000 rpm, 25 °C). The precipitated fraction was solubilized in water, followed by freezing and thawing (3x), giving rise to soluble and insoluble fractions. The soluble fraction was dissolved in water and underwent ultrafiltration using a 3 kDa cut-off membrane (Millipore®; polyethersulfone membrane) coupled in a Sartorius cylinder (Model 16249). The retained fraction was characterized as a (1→3),(1→6) and (1→3)-β-D-glucan named PN (Table 1), which was later used for in vitro fecal fermentation experiments.

[0046] P. pulmonarius glucan (PP) – The lyophilized fruiting bodies of the mushroom P. pulmonarius were ground and dissolved in distilled water (0.3 L / 10 g), reduced with a small aliquot of NaBH4and submitted to high temperature and pressure extraction (121 °C, 1.2 atm, 1 hour) using an autoclave. The materials were filtered, and the retained fraction was subjected to a new autoclave extraction, now in an alkaline medium with 5% potassium hydroxide, and then centrifuged (20 minutes, 10,000 rpm, 25 °C). The supernatant was then neutralized, dialyzed and precipitated with ethanol (3:1, v / v) followed by centrifugation (20 minutes, 10,000 rpm, 25 °C). The precipitated fraction was solubilized in water, followed by freezing and thawing (3x), giving rise to soluble and insoluble fractions. The insoluble fraction was characterized as a (1→3),(1→6) and (1→3)-β-D-glucan named PP (Table 1), which was later used for in vitro fecal fermentation experiments.

[0047] A previously obtained (1→3),(1→6) and (1→3)-β-D-glucan from Cookeina speciosa (CS) was used as a positive control for promotion of Anaerostipes sp. and B. uniformis (Cantu-Jungles et al., 2021), and fructooligosaccharides (FOS - No. F8052, Sigma-Aldrich Inc.,70085-02 St. Louis, Mo., USA) as a fermentable negative control for promotion of Anaerostipes sp. and B. uniformis. Example 1 Modification of P. pulmonarius and P. nameko β-glucans structures

[0048] To investigate the importance of the glucan side chains to gut microbiota shifts, glucans obtained from P. pulmonarius and P. nameko were each subjected to a Smith degradation process, resulting in debranched glucans, which were further used for in vitro fecal fermentation experiments. Briefly, an aliquot of each purified β-D-glucan (500 mg) obtained from P. nameko and P. pulmonarius was submitted to 0.05 M aq. NaIO4 (20 mL) for 72 hours at 25 °C in the dark under stirring using a magnetic bar (Delgobo et al., 1998). The oxidation process was stopped with ethylene glycol (2 mL), and each sample was then dialyzed against tap water for 12 hours, concentrated, reduced with NaBH4and maintained overnight at room temperature. The process was stopped with acetic acid followed by dialysis (2 kDa, 24 hours) and freeze drying. The products of the respective polyalcohols were hydrolyzed with TFA 1M (4.0 mL, 30 min, 100 °C) and dialyzed (2 kDa) against tap water for 12 hours and lyophilized. The residual polysaccharides were characterized as linear (1→3)-β-D-glucans (Table 1) and named as PP-Sm (linear glucan from P. pulmonarius) and PN-Sm (linear glucan from P. nameko). Example 2 Linkage analysis of GT, GL, PN, PP, PN-Sm and PP-Sm glucans

[0049] The linkage patterns of GT, GL, PN, PP, PN-Sm and PP-Sm β-D-glucans were evaluated through methylation analysis. Briefly, using a method modified from Ciucanu and Kerek (1984), per-O-methylation of the purified fractions (10 mg) was carried out using NaOH-Me2SO-MeI as described by Ruthes et al (2010). The per-O- methylated derivatives were hydrolyzed with TFA 2M (1 mL,100 °C, 20 hours) followed by evaporation to dryness and reduction with NaBD4. Then, samples were acetylated and converted into partially O-methylated alditol acetates and analyzed by GC-MS using He as the carrier gas at a flow rate of 1.0 mL min-1. Partially O-methylated alditol acetates70085-02 were identified by the ion m / z by comparing their positive ions with standards. The results are expressed as a relative percentage of each component (Sassaki et al., 2005).

[0050] Methylation data confirmed that fractions GT, GL, PN, and PP consisted of branched (1→3),(1→6) and (1→3)-β-D-glucans, each with different proportions in linkage types present, while GL also presented 6% of (1→4)-D- linked glucose units (Table 1). Moreover, the Smith degradation procedure effectively removed branches from PN and PP, as observed in Table 1 from the absence of 6→)-Glcp-(1→ and 3,6→)-Glcp-(1→ methylation derivatives in fractions PN-Sm and PP-SM. Table 1. Partially O-methylated acetates present on D-glucans (GT, GL, PN, PP, PN-Sm and PP-Sm) obtained from different mushroom sources Partially O- % area of fragmentsbLinkage types methylated PPcPNdPPSmePNSmfGTgGLhβ-D-can- (1→3) from P. pulmonarius;fPNSm: β-D-glucan-(1→3) from P. nameko,gGT: β-D-glucan-(1→3),(1→6) from G. thelephoroides;hGL: β-D-glucan-(1→3),(1→6) from G. lucidum. Example 3 In vitro fecal fermentation profile by the human gut microbiota

[0051] Glucans GT, GL, PN, and PP and the debranched glucans PN-Sm and PP-Sm were submitted to in vitro fecal fermentations, as well as the positive control CS and negative controls FOS (fermentable fiber) and blank (no fiber added) as previously described (Cantu- Jungles et al., 2021). Each substrate (50^mg equivalent carbohydrate, as determined above by the phenol-sulfuric acid method) was weighed in three test tubes for triplicate analysis. Hydration of samples was performed by adding 4^mL of a carbonate-phosphate buffer, pH 6.8^±^0.1, to each tube. Fructooligosaccharides (FOS - No. F8052, Sigma-Aldrich Inc., St. Louis, Mo., USA) and tubes without any added carbohydrate were used as positive and negative controls, respectively. Fecal samples were obtained from three healthy volunteers, who were on their routine diet and70085-02 had not taken antibiotics within the previous six months. Fecal samples were collected in plastic bags, which were sealed after removing air, transported on ice, and immediately placed inside an anaerobic chamber (10% H2, 5% CO2, and 85% N2; BactronEZ, SHEL LAB, Cornelius, OR), where all further procedures were performed within two^hours after collection. Fecal samples were pooled together, and the fecal slurry was prepared by homogenization with carbonate- phosphate buffer, pH 6.8^±^0.1, in a ratio of 1:3 (w / v) and further strained through four layers of cheesecloth. The filtrate (1^mL) was then inoculated with the hydrated carbohydrate sample and the controls. Tubes were closed, sealed, and incubated at 37°C in a shaker incubator (150^rpm; MaxQ 6000; Thermo Fisher, Waltham, MA) for 24 hours. Aliquots of the baseline sample and samples after 24-hour fermentations were prepared and stored at −80°C until further use for SCFA analysis (0.5^ml) and DNA sequencing (1^ml). Human stool collection and use were approved by the Institutional Review Board at Purdue University (IRB protocol no. 1510016635). Example 4 Short chain fatty acids (SCFAs) quantification

[0052] Samples for SCFA analyses were prepared and analyzed using a gas chromatograph (GC-FID 7890^A; Agilent Technologies Inc.) on a fused silica capillary column (Nukon Supelco no.40369-03A; Bellefonte, PA) as previously described (Cantu-Jungles et al., 2018). Quantification was performed based on relative peak area using external standards of acetate (A38S), propionate (A258), and butyrate (AC108111000) and an internal standard of 4- methylvaleric acid (AAA1540506) from Fisher Scientific (Hampton, NH).

[0053] Fermentation of all samples increased total SCFA production compared to the blanks, and PN and GT led to the highest total SCFA production (95.2 mM and 97 mM, respectively) across tested glucans (Fig.1A). Branching removal of PN that resulted in the insoluble PNSm fraction reduced its SCFA production to 61.4 mM. Whereas lower than PN and GT, other glucans still produced at least 40% of the total SCFAs amount produced by the positive control FOS, indicating that, despite their insolubility, a good portion was still fermented (Fig.1A). Regarding production of each SCFA, acetate was higher across tested glucans for GT and PN (47.6 and 40.0 mM, respectively, Fig.1B), and propionate reached the highest levels in PN (43.03 mM), while still produced in good amounts (20.9 -27.0 mM) for all tested glucans70085-02 (Figs.1B and 1C). For butyrate, CS, GT and GL were the most butyrogenic across tested glucans and, while PP and PN led to similar butyrate production, their debranched counterparts (PPSm and PNSm) significantly reduced butyrate levels (Figs.1E and 1F).

[0054] Evaluation of SCFAs proportions from the total SCFA amount produced was useful to determine which SCFA bacterial types were promoted during fermentation. All glucans led to a high proportion of propionate compared to the controls (Fig.1E). For butyrate, the CS, GT, GL and PP fractions presented the highest proportions across tested samples (Fig.1E). Example 5 DNA extraction, 16S rRNA gene amplicon sequencing

[0055] Stored samples for DNA extraction were thawed and centrifuged (13,000^rpm for 15^min), and supernatants were discarded. Automated DNA extraction of the precipitates was performed using the QIAcube Connect instrument (Qiagen, Germantown, MD) with the QIAamp PowerFecal Pro DNA kit (Qiagen, Germantown, MD) as per manufacturer’s instructions. The V4 region of 16S rRNA gene was amplified using primers 515F 5'- GTGCCAGCMGCCGCGGTAA [SEQ ID NO: 10] and 806R 5'- GGACTACHVHHHTWTCTAAT [SEQ ID NO: 11] and then sequenced using the Illumina MiniSeq platform (Illumina, Inc., San Diego, CA). Library preparation and 16S rRNA gene sequencing were performed at the Rush Genomics and Microbiome Core Facility (Chicago, IL).

[0056] Shifts in gut microbial communities were evaluated through 16S rRNA gene amplicon sequencing. ANCOM analysis identified five bacteria taxa that were differentially promoted across one or more samples: Anaerostipes sp., B. uniformis, Bacteroides ovatus, Roseburia sp., and Parabacteroides distasonis (Fig.2).

[0057] The most promoted bacteria with the tested, non-modified glucans was Anaerostipes sp. Anaerostipes sp. was promoted by GT (22%) and GL (21.8%) at a level similar to the previously isolated CS (20.9%) and also significantly promoted by PP (12.9%) and PN (4.7%) in comparison to the blanks (0.3 - 0.4%) and FOS (0.05%, Fig.2A). Interestingly, debranching of PP and PN completely prevented Anaerostipes sp. promotion, indicating that the branching structure is important for promotion of this taxa (Figs.2A and 4).

[0058] Anaerostipes sp. was better promoted with the insoluble GT, GL, CS and the insoluble branched PP than the soluble PN. Taking together, these support the idea that both70085-02 branching and solubility of mushroom β-D-glucans are important for promotion of Anaerostipes sp. Anaerostipes is a genus of the human large intestine with several butyrate producers, with functions related to local and systemic human health (Bui et al., 2014; Canani et al., 2011; Cantu-Jungles et al., 2018; Kant et al., 2015; Lee et al., 2021; Schwiertz et al., 2002). This corroborates the observation that butyrate proportions were higher in glucans more promotive of Anaerostipes sp. (Fig.1E).

[0059] The second most promoted bacterium was B. uniformis, which corresponded to 1.9% of the initial community (blank, 0 hour) and reached concentrations ranging from 9.3% to 18.8% after fermentation of glucans. B. uniformis has been previously shown to ameliorate the immunological dysfunctions and metabolic disorders in animal models with suggested roles in obesity prevention (Benítez-Páez et al., 2017; Wang et al., 2021). B. uniformis also produces propionate (Hosseini Jazani and Shahabi, 2019; Rios-Covian et al., 2017), a metabolite closely related to health. In fact, high propionate proportions were observed in all tested glucans (Fig. 2E).

[0060] Other beneficial bacteria were promoted to a lower extent and were different depending on the glucan tested. Bacteroide ovatus was promoted by FOS, PP and PN, but not by their debranched counterparts (PPSm and PNSm, Fig.2D). Roseburia sp., another butyrate producer, was especially promoted by GL (from 1.7% in Blank, 0 hour, to 6.9% in GL). P. distasonis was mostly promoted in PN (6.1%) and GT (5.3%). Interestingly, whereas debranching of PP into PPSm increased P. distasonis abundance from 1.7 to 3.3%, debranching of PN into PNSm reduced its abundance from 6.1 to 3.8 %, indicating that the glucan branching degree is not determinant for P. distasonis growth support (Fig.2E). Example 6 Bioinformatics

[0061] Demultiplexed and preprocessed sequence reads were supplied as paired-end FASTQ sequence files and imported into QIIME 2 (q2) version 2019-10 for analysis (Bolyen et al., 2019). Amplicon sequence variants (ASVs) were generated using DADA2 (Callahan et al., 2016) (sequences were trimmed at 153 bp), and taxonomic assignment was carried out using the q2-feature-classifier plugin against the Greengenes reference database classifier with 99% similarity, specific for the V416S region (v.3.8). Sequence alignment and construction of a70085-02 phylogeny tree were generated using the Qiime2 pipeline align-to-tree-mafft-fasttree. Relative abundances were generated with the q2-taxa plugin with ASVs collapsed at the species level for differentially abundant bacterial taxa. Differentially abundant species across the 15 most abundant taxa were selected using analysis of composition of microbiomes (ANCOM) through the q2-composition plugin, and displayed as relative abundances (Mandal et al., 2015). Short chain fatty acids before and after 24 hour in vitro fecal fermentation were analyzed through one- way ANOVA followed by Tukey’s post-hoc test. Selected differentially abundant species and SCFAs data were plotted and analyzed using GraphPad prism (v.9.1.2).

[0062] Overall, GT GL PP and PN glucan led to targeted promotion of Anaerostipes sp. and B. uniformis. These are precisely the same bacterial taxa previously found to be consistently promoted in different individuals using an insoluble (1→3),(1→6) β-D-glucan from Cookeina speciosa mushroom (Cantu-Jungles et al., 2018). Other beneficial bacteria, which were promoted by some of the glucans to a lower extent, were B. ovatus, Roseburia sp., and P. distasonis. Some of the gut microbial outcomes and metabolites largely relied on the β-D-glucans physico- chemical structural features. This is in agreement with previous hypotheses and studies that showed that even small shifts in physico-chemical structure of the same class of polymer could differentiate their action towards the human gut microbial communities and health-related outcomes (Deehan et al., 2020; Hamaker and Tuncil, 2014; Zhang et al., 2019, 2021). 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[0063] All patents, patent application publications, journal articles, textbooks, and other publications mentioned in the specification are indicative of the level of skill of those in the art to which the disclosure pertains. All such publications are incorporated herein by reference to the same extent as if each individual publication were specifically and individually indicated to be incorporated by reference. In the event of inconsistent usages between this document and those documents so incorporated by reference, the usage in the incorporated reference should be70085-02 considered supplementary to that of this document; for irreconcilable inconsistencies, the usage in this document controls.

[0064] The invention illustratively described herein may be suitably practiced in the absence of any element(s) or limitation(s), which is / are not specifically disclosed herein. Thus, for example, each instance herein of any of the terms "comprising," "consisting essentially of," and "consisting of" may be replaced with either of the other two terms. Likewise, the singular forms "a," "an," and "the" include plural references unless the context clearly dictates otherwise. Thus, for example, references to "the method" includes one or more methods and / or steps of the type, which are described herein and / or which will become apparent to those ordinarily skilled in the art upon reading the disclosure. The term "or" is used to refer to a nonexclusive "or" unless otherwise indicated.

[0065] Unless defined otherwise, all technical and scientific terms used herein have the same meaning as commonly understood to one of ordinary skill in the art. The following terms and phrases shall have the meaning indicated.

[0066] The term "about," when referring to a number or a numerical value or range (including, for example, whole numbers, fractions, and percentages), means that the number or numerical range referred to is an approximation within experimental variability (or within statistical experimental error) and thus the numerical value or range can vary between 1% and 15% of the stated number or numerical range (e.g., + / - 5 % to 15% of the recited value, such as within 10%, within 5%, or within 1% of a stated value or stated limit of a range) provided that one of ordinary skill in the art would consider equivalent to the recited value (e.g., having the same function or result). The term "substantially" can allow for a degree of variability in a value or range, for example, within 90%, within 95%, 99%, 99.5%, 99.9%, 99.99%, or at least about 99.999% or more of a stated value or of a stated limit of a range.

[0067] In addition, it is to be understood that the phraseology or terminology employed herein, and not otherwise defined, is for the purpose of description only and not of limitation. Any use of section headings is intended to aid reading of the document and is not to be interpreted as limiting. Further, information that is relevant to a section heading may occur within or outside of that particular section.70085-02 ENUMERATED EMBODIMENTS 1. A prebiotic fiber composition obtained from Gloeoporus thelephoroides (GT) by a method comprising: (i) grinding, or obtaining ground, lyophilized fruiting bodies of GT; (ii) dissolving the ground, lyophilized fruiting bodies of GT in distilled water, or in lieu of steps (i) and (ii), obtaining ground, lyophilized fruiting bodies of GT dissolved in distilled water and optionally reduced with NaBH4; (iii) if not already reduced, reducing the ground, lyophilized fruiting bodies of GT dissolved in distilled water with NaBH4; (iv) subjecting the reduced, ground, lyophilized fruiting bodies of GT dissolved in distilled water to exhaustive hot-water extraction; (v) filtering the extraction to obtain a filtrate; (vi) subjecting the filtrate of (v) to extraction in an alkaline medium; (vii) centrifuging the extraction of (vi) to obtain a supernatant; (viii) neutralizing, dialyzing and precipitating the supernatant of (vii); (ix) centrifuging the precipitated supernatant of (viii) to obtain a precipitate; and (x) lyophilizing the precipitate of (ix) or solubilizing the precipitate of (ix) in water, whereupon the prebiotic fiber composition from GT is obtained. 2. The prebiotic fiber composition of claim 1, which comprises (1→3),(1→6) and (1→3)-β-D-glucan. 3. The prebiotic fiber composition of claim 1 or 2, wherein exhaustive hot-water extraction is carried out for about 3 hours (e.g., 3 hours) at about 100°C (e.g., 100°C) using a water bath under reflux. 4. The prebiotic fiber composition of any one of claims 1-3, wherein the alkaline medium of (vi) comprises 5% potassium hydroxide.70085-02 5. The prebiotic fiber composition of any one of claims 1-4, wherein the centrifuging in (vii) and (ix) is carried out at about 10,000 rpm (e.g., 10,000 rpm) at about 25 °C (e.g., 25 °C) for about 20 minutes (e.g., 20 minutes). 6. The prebiotic fiber composition of any one of claims 1-5, wherein the precipitating in (viii) is carried out in ethanol (3:1 v / v). 7. A prebiotic fiber composition obtained from Gloeoporus thelephoroides (GT), which comprises (1→3),(1→6) and (1→3)-β-D-glucan. 8. The prebiotic fiber composition of claim 7, which is lyophilized. 9. The prebiotic fiber composition of claim 7, which is insoluble or solubilized in water. 10. A method of promoting growth of Anaerostipes sp., Bacteroides uniformis, or both in the gut microbiome of an animal (e.g., warm-blooded animal, e.g., mammal, e.g., human), which method comprises orally administering the prebiotic fiber composition of any one of claims 1-9, alone or in further combination with live Anaerostipes sp., Bacteroides uniformis, or both, simultaneously (e.g., as one, two or three compositions) or sequentially (e.g., as two or three compositions), whereupon growth of Anaerostipes sp., Bacteroides uniformis, or both in the gut microbiome of the animal is promoted. 11. A prebiotic fiber composition obtained from Ganoderma lucidum (GL) by a method comprising: (i) grinding, or obtaining ground, lyophilized fruiting bodies of GL; (ii) dissolving the ground, lyophilized fruiting bodies of GL in distilled water, or in lieu of steps (i) and (ii), obtaining ground, lyophilized fruiting bodies of GL dissolved in distilled water and optionally reduced with NaBH4; (iii) if not already reduced, reducing the ground, lyophilized fruiting bodies of GL dissolved in distilled water with NaBH4;70085-02 (iv) subjecting the reduced, ground, lyophilized fruiting bodies of GL dissolved in distilled water to exhaustive hot-water extraction; (v) filtering the extraction to obtain a filtrate; (vi) subjecting the filtrate of (v) to extraction in an alkaline medium; (vii) centrifuging the extraction of (vi) to obtain a supernatant; (viii) neutralizing, dialyzing and precipitating the supernatant of (vii); (ix) centrifuging the precipitated supernatant of (viii) to obtain a precipitate; and (x) lyophilizing the precipitate of (ix) or solubilizing the precipitate of (ix) in water, whereupon the prebiotic fiber composition from GL is obtained. 12. The prebiotic fiber composition of claim 11, which comprises (1→3),(1→6) and (1→3)-β-D-glucan. 13. The prebiotic fiber composition of claim 11 or 12, wherein exhaustive hot- water extraction is carried out for about 3 hours (e.g., 3 hours) at about 100°C (e.g., 100°C) using a water bath under reflux. 14. The prebiotic fiber composition of any one of claims 11-13, wherein the alkaline medium of (vi) comprises 5% potassium hydroxide. 15. The prebiotic fiber composition of any one of claims 11-14, wherein the centrifuging in (vii) and (ix) is carried out at about 10,000 rpm (e.g., 10,000 rpm) at about 25 °C (e.g., 25 °C) for about 20 minutes (e.g., 20 minutes). 16. The prebiotic fiber composition of any one of claims 11-15, wherein the precipitating in (viii) is carried out in ethanol (3:1 v / v). 17. A prebiotic fiber composition obtained from Ganoderma lucidum (GL), which comprises (1→3),(1→6) and (1→3)-β-D-glucan. 18. The prebiotic fiber composition of claim 17, which is lyophilized.70085-02 19. The prebiotic fiber composition of claim 17, which is insoluble or solubilized in water. 20. A method of promoting growth of Anaerostipes sp., Bacteroides uniformis, or both in the gut microbiome of an animal (e.g., warm-blooded animal, e.g., mammal, e.g., human), which method comprises orally administering the prebiotic fiber composition of any one of claims 11- 19, alone or in further combination with live Anaerostipes sp., Bacteroides uniformis, or both, simultaneously (e.g., as one, two or three compositions) or sequentially (e.g., as two or three compositions), whereupon growth of Anaerostipes sp., Bacteroides uniformis, or both in the gut microbiome of the animal is promoted. 21. A prebiotic fiber composition obtained from Pholiota nameko (PN) by a method comprising: (i) grinding, or obtaining ground, lyophilized fruiting bodies of PN; (ii) dissolving the ground, lyophilized fruiting bodies of PN in distilled water, or in lieu of steps (i) and (ii), obtaining ground, lyophilized fruiting bodies of PN dissolved in distilled water and optionally reduced with NaBH4; (iii) if not already reduced, reducing the ground, lyophilized fruiting bodies of PN dissolved in distilled water with NaBH4; (iv) subjecting the reduced, ground, lyophilized fruiting bodies of PN dissolved in distilled water to extraction under high temperature and high pressure to obtain an extract; (v) filtering the extract of (iv) to obtain an eluant; (vi) precipitating the eluant from (v) to obtain a precipitated supernatant; (vii) centrifuging the precipitated supernatant of (vi); (viii) solubilizing the precipitate of (vii) in water; (ix) freeze-thawing the solubilized precipitate of (viii) three times to obtain soluble and insoluble fractions; (x) dissolving the soluble fraction of (ix) in water; (xi) ultra-filtering the dissolved soluble fraction of (x) using a membrane with a 3kDa cut-off, and70085-02 (xii) lyophilizing the filtrate of (ix) or solubilizing the filtrate of (ix) in water, whereupon the prebiotic fiber composition from PN is obtained. 22. The prebiotic fiber composition of claim 21, which comprises (1→3),(1→6) and (1→3)-β-D-glucan. 23. The prebiotic fiber composition of claim 21 or 22, wherein extraction under high temperature and high pressure is carried out for about one hour (one hour) at about 121 °C (e.g., 121 °C) and about 1.2 atm (e.g., 1.2 atm) using an autoclave. 24. The prebiotic fiber composition of any one of claims 21-23, wherein the precipitating in (vi) is carried out in ethanol (3:1 v / v). 25. The prebiotic fiber composition of any one of claims 21-24, wherein the centrifuging in (vii) is carried out at about 10,000 rpm (e.g., 10,000 rpm) at about 25 °C (e.g., 25 °C) for about 20 minutes (e.g., 20 minutes). 26. A prebiotic fiber composition obtained from Pholiota nameko (PN), which comprises (1→3),(1→6) and (1→3)-β-D-glucan. 27. The prebiotic fiber composition of claim 26, which is lyophilized. 28. The prebiotic fiber composition of claim 26, which is insoluble or solubilized in water. 29. A method of promoting growth of Anaerostipes sp., Bacteroides uniformis, or both in the gut microbiome of an animal (e.g., warm-blooded animal, e.g., mammal, e.g., human), which method comprises orally administering the prebiotic fiber composition of any one of claims 21-28, alone or in further combination with live Anaerostipes sp., Bacteroides uniformis, or both, simultaneously (e.g., as one, two or three compositions)70085-02 or sequentially (e.g., as two or three compositions), whereupon growth of Anaerostipes sp., Bacteroides uniformis, or both in the gut microbiome of the animal is promoted. 30. A prebiotic fiber composition obtained from Pleurotus pulmonarius (PP) by a method comprising: (i) grinding, or obtaining ground, lyophilized fruiting bodies of PP; (ii) dissolving the ground, lyophilized fruiting bodies of PP in distilled water, or in lieu of steps (i) and (ii), obtaining ground, lyophilized fruiting bodies of PP dissolved in distilled water and optionally reduced with NaBH4; (iii) if not already reduced, reducing the ground, lyophilized fruiting bodies of PP dissolved in distilled water with NaBH4; (iv) subjecting the reduced, ground, lyophilized fruiting bodies of PN dissolved in distilled water to extraction under high temperature and high pressure; (v) filtering the extraction to obtain a filtrate; (vi) extracting the filtrate of (v) in an alkaline medium to obtain an extraction; (vii) centrifuging the extraction of (vi) to obtain a supernatant; (viii) neutralizing, dialyzing and precipitating the supernatant; (ix) centrifuging the precipitated supernatant of (viii) to obtain a precipitate; (x) solubilizing the precipitate of (ix) in water; (xi) freeze-thawing the solubilized precipitate of (x) three times to obtain soluble and insoluble fractions; and (xii) lyophilizing the precipitate of (ix) or solubilizing the precipitate of (ix) in water, whereupon the prebiotic fiber composition from PP is obtained. 31. The prebiotic fiber composition of claim 30, which comprises (1→3),(1→6) and (1→3)-β-D-glucan. 32. The prebiotic fiber composition of claim 30 or 31, wherein extraction under high temperature and high pressure is carried out for about one hour (e.g., one hour) at about 121 °C (e.g., 121 °C) and about 1.2 atm (e.g., 1.2 atm) using an autoclave.70085-02 33. The prebiotic fiber composition of any one of claims 30-32, wherein the alkaline medium of (vi) comprises 5% potassium hydroxide. 34. The prebiotic fiber composition of any one of claims 30-33, wherein the centrifuging in (vii) and (ix) is carried out at about 10,000 rpm (e.g., 10,000 rpm) at about 25 °C (e.g., 25 °C) for about 20 minutes (e.g., 20 minutes). 35. The prebiotic fiber composition of any one of claims 30-34, wherein the precipitating in (viii) is carried out in ethanol (3:1 v / v). 36. A prebiotic fiber composition obtained from Pleurotus pulmonarius (PP), which comprises (1→3),(1→6) and (1→3)-β-D-glucan. 37. The prebiotic fiber composition of claim 36, which is lyophilized. 38. The prebiotic fiber composition of claim 36, which is insoluble or solubilized in water. 39. A method of promoting growth of Anaerostipes sp., Bacteroides uniformis, or both in the gut microbiome of an animal (e.g., warm-blooded animal, e.g., mammal, e.g., human), which method comprises orally administering the prebiotic fiber composition of any one of claims 30- 38, alone or in further combination with live Anaerostipes sp., Bacteroides uniformis, or both, simultaneously (e.g., as one, two or three compositions) or sequentially (e.g., as two or three compositions), whereupon growth of Anaerostipes sp., Bacteroides uniformis, or both in the gut microbiome of the animal is promoted. 40. A prebiotic fiber composition comprising: (a) a prebiotic fiber composition obtained from Gloeoporus thelephoroides (GT), which comprises (1→3),(1→6) and (1→3)-β-D-glucan; (b) a prebiotic fiber composition obtained from Ganoderma lucidum (GL), which comprises (1→3),(1→6) and (1→3)-β-D-glucan;70085-02 (c) a prebiotic fiber composition obtained from Pholiota nameko (PN), which comprises (1→3),(1→6) and (1→3)-β-D-glucan; (d) a prebiotic fiber composition obtained from Pleurotus pulmonarius (PP), which comprises (1→3),(1→6) and (1→3)-β-D-glucan; (e) (a) and (b); (f) (a) and (c); (g) (a) and (d); (h) (b) and (c); (i) (b) and (d); (j) (c) and (d); or (k) (a), (b), (c), and (d). 41. The prebiotic fiber composition of claim 40, which is lyophilized. 42. The prebiotic fiber composition of claim 40, which is insoluble or solubilized in water. 43. A method of promoting growth of Anaerostipes sp., Bacteroides uniformis, or both in the gut microbiome of an animal (e.g., warm-blooded animal, e.g., mammal, e.g., human), which method comprises orally administering the prebiotic fiber composition of any one of claims 40- 42, alone or in further combination with live Anaerostipes sp., Bacteroides uniformis, or both, simultaneously (e.g., as one or more compositions) or sequentially (e.g., as one or more compositions), whereupon growth of Anaerostipes sp., Bacteroides uniformis, or both in the gut microbiome of the animal is promoted.

Claims

70085-02 WHAT IS CLAIMED IS:

1. A prebiotic fiber composition obtained from Gloeoporus thelephoroides (GT) by a method comprising: (i) grinding, or obtaining ground, lyophilized fruiting bodies of GT; (ii) dissolving the ground, lyophilized fruiting bodies of GT in distilled water, or in lieu of steps (i) and (ii), obtaining ground, lyophilized fruiting bodies of GT dissolved in distilled water and optionally reduced with NaBH4; (iii) if not already reduced, reducing the ground, lyophilized fruiting bodies of GT dissolved in distilled water with NaBH4; (iv) subjecting the reduced, ground, lyophilized fruiting bodies of GT dissolved in distilled water to exhaustive hot-water extraction; (v) filtering the extraction to obtain a filtrate; (vi) subjecting the filtrate of (v) to extraction in an alkaline medium; (vii) centrifuging the extraction of (vi) to obtain a supernatant; (viii) neutralizing, dialyzing and precipitating the supernatant of (vii); (ix) centrifuging the precipitated supernatant of (viii) to obtain a precipitate; and (x) lyophilizing the precipitate of (ix) or solubilizing the precipitate of (ix) in water, whereupon the prebiotic fiber composition from GT is obtained.

2. The prebiotic fiber composition of claim 1, which comprises (1→3),(1→6) and (1→3)-β-D-glucan.

3. The prebiotic fiber composition of claim 1, wherein exhaustive hot-water extraction is carried out for about 3 hours at about 100°C using a water bath under reflux.

4. The prebiotic fiber composition of claim 1, wherein the alkaline medium of (vi) comprises 5% potassium hydroxide.

5. The prebiotic fiber composition of claim 1, wherein the centrifuging in (vii) and (ix) is carried out at about 10,000 rpm at about 25 °C for about 20 minutes.70085-02 6. The prebiotic fiber composition of claim 1, wherein the precipitating in (viii) is carried out in ethanol (3:1 v / v).

7. A prebiotic fiber composition obtained from Gloeoporus thelephoroides (GT), which comprises (1→3),(1→6) and (1→3)-β-D-glucan.

8. The prebiotic fiber composition of claim 7, which is lyophilized.

9. The prebiotic fiber composition of claim 7, which is insoluble or solubilized in water.

10. A method of promoting growth of Anaerostipes sp., Bacteroides uniformis, or both in the gut microbiome of an animal, which method comprises orally administering the prebiotic fiber composition of claim 1, alone or in further combination with live Anaerostipes sp., Bacteroides uniformis, or both, simultaneously or sequentially, whereupon growth of Anaerostipes sp., Bacteroides uniformis, or both in the gut microbiome of the animal is promoted.

11. A prebiotic fiber composition obtained from Ganoderma lucidum (GL) by a method comprising: (i) grinding, or obtaining ground, lyophilized fruiting bodies of GL; (ii) dissolving the ground, lyophilized fruiting bodies of GL in distilled water, or in lieu of steps (i) and (ii), obtaining ground, lyophilized fruiting bodies of GL dissolved in distilled water and optionally reduced with NaBH4; (iii) if not already reduced, reducing the ground, lyophilized fruiting bodies of GL dissolved in distilled water with NaBH4; (iv) subjecting the reduced, ground, lyophilized fruiting bodies of GL dissolved in distilled water to exhaustive hot-water extraction; (v) filtering the extraction to obtain a filtrate; (vi) subjecting the filtrate of (v) to extraction in an alkaline medium; (vii) centrifuging the extraction of (vi) to obtain a supernatant; (viii) neutralizing, dialyzing and precipitating the supernatant of (vii);70085-02 (ix) centrifuging the precipitated supernatant of (viii) to obtain a precipitate; and (x) lyophilizing the precipitate of (ix) or solubilizing the precipitate of (ix) in water, whereupon the prebiotic fiber composition from GL is obtained.

12. The prebiotic fiber composition of claim 11, which comprises (1→3),(1→6) and (1→3)-β-D-glucan.

13. The prebiotic fiber composition of claim 11, wherein exhaustive hot-water extraction is carried out for about 3 hours at about 100°C using a water bath under reflux.

14. The prebiotic fiber composition of claim 11, wherein the alkaline medium of (vi) comprises 5% potassium hydroxide.

15. The prebiotic fiber composition of claim 11, wherein the centrifuging in (vii) and (ix) is carried out at about 10,000 rpm at about 25 °C for about 20 minutes.

16. The prebiotic fiber composition of claim 11, wherein the precipitating in (viii) is carried out in ethanol (3:1 v / v).

17. A prebiotic fiber composition obtained from Ganoderma lucidum (GL), which comprises (1→3),(1→6) and (1→3)-β-D-glucan.

18. The prebiotic fiber composition of claim 17, which is lyophilized.

19. The prebiotic fiber composition of claim 17, which is insoluble or solubilized in water.

20. A method of promoting growth of Anaerostipes sp., Bacteroides uniformis, or both in the gut microbiome of an animal, which method comprises orally administering the prebiotic fiber composition of claim 11, alone or in further combination with live Anaerostipes sp., Bacteroides uniformis, or both, simultaneously or sequentially,70085-02 whereupon growth of Anaerostipes sp., Bacteroides uniformis, or both in the gut microbiome of the animal is promoted.

21. A prebiotic fiber composition obtained from Pholiota nameko (PN) by a method comprising: (i) grinding, or obtaining ground, lyophilized fruiting bodies of PN; (ii) dissolving the ground, lyophilized fruiting bodies of PN in distilled water, or in lieu of steps (i) and (ii), obtaining ground, lyophilized fruiting bodies of PN dissolved in distilled water and optionally reduced with NaBH4; (iii) if not already reduced, reducing the ground, lyophilized fruiting bodies of PN dissolved in distilled water with NaBH4; (iv) subjecting the reduced, ground, lyophilized fruiting bodies of PN dissolved in distilled water to extraction under high temperature and high pressure to obtain an extract; (v) filtering the extract of (iv) to obtain an eluant; (vi) precipitating the eluant from (v) to obtain a precipitated supernatant; (vii) centrifuging the precipitated supernatant of (vi); (viii) solubilizing the precipitate of (vii) in water; (ix) freeze-thawing the solubilized precipitate of (viii) three times to obtain soluble and insoluble fractions; (x) dissolving the soluble fraction of (ix) in water; (xi) ultra-filtering the dissolved soluble fraction of (x) using a membrane with a 3kDa cut-off, and (xii) lyophilizing the filtrate of (ix) or solubilizing the filtrate of (ix) in water, whereupon the prebiotic fiber composition from PN is obtained.

22. The prebiotic fiber composition of claim 21, which comprises (1→3),(1→6) and (1→3)-β-D-glucan.

23. The prebiotic fiber composition of claim 21, wherein extraction under high temperature and high pressure is carried out for about one hour at about 121 °C and about 1.2 atm using an autoclave.70085-02 24. The prebiotic fiber composition of claim 21, wherein the precipitating in (vi) is carried out in ethanol (3:1 v / v).

25. The prebiotic fiber composition of claim 21, wherein the centrifuging in (vii) is carried out at about 10,000 rpm at about 25 °C for about 20 minutes.

26. A prebiotic fiber composition obtained from Pholiota nameko (PN), which comprises (1→3),(1→6) and (1→3)-β-D-glucan.

27. The prebiotic fiber composition of claim 26, which is lyophilized.

28. The prebiotic fiber composition of claim 26, which is insoluble or solubilized in water.

29. A method of promoting growth of Anaerostipes sp., Bacteroides uniformis, or both in the gut microbiome of an animal, which method comprises orally administering the prebiotic fiber composition of claim 21, alone or in further combination with live Anaerostipes sp., Bacteroides uniformis, or both, simultaneously or sequentially, whereupon growth of Anaerostipes sp., Bacteroides uniformis, or both in the gut microbiome of the animal is promoted.

30. A prebiotic fiber composition obtained from Pleurotus pulmonarius (PP) by a method comprising: (i) grinding, or obtaining ground, lyophilized fruiting bodies of PP; (ii) dissolving the ground, lyophilized fruiting bodies of PP in distilled water, or in lieu of steps (i) and (ii), obtaining ground, lyophilized fruiting bodies of PP dissolved in distilled water and optionally reduced with NaBH4; (iii) if not already reduced, reducing the ground, lyophilized fruiting bodies of PP dissolved in distilled water with NaBH4;70085-02 (iv) subjecting the reduced, ground, lyophilized fruiting bodies of PN dissolved in distilled water to extraction under high temperature and high pressure; (v) filtering the extraction to obtain a filtrate; (vi) extracting the filtrate of (v) in an alkaline medium to obtain an extraction; (vii) centrifuging the extraction of (vi) to obtain a supernatant; (viii) neutralizing, dialyzing and precipitating the supernatant; (ix) centrifuging the precipitated supernatant of (viii) to obtain a precipitate; (x) solubilizing the precipitate of (ix) in water; (xi) freeze-thawing the solubilized precipitate of (x) three times to obtain soluble and insoluble fractions; and (xii) lyophilizing the precipitate of (ix) or solubilizing the precipitate of (ix) in water, whereupon the prebiotic fiber composition from PP is obtained.

31. The prebiotic fiber composition of claim 30, which comprises (1→3),(1→6) and (1→3)-β-D-glucan.

32. The prebiotic fiber composition of claim 30, wherein extraction under high temperature and high pressure is carried out for about one hour at about 121 °C and about 1.2 atm using an autoclave.

33. The prebiotic fiber composition of claim 30, wherein the alkaline medium of (vi) comprises 5% potassium hydroxide.

34. The prebiotic fiber composition of claim 30, wherein the centrifuging in (vii) and (ix) is carried out at about 10,000 rpm at about 25 °C for about 20 minutes.

35. The prebiotic fiber composition of claim 30, wherein the precipitating in (viii) is carried out in ethanol (3:1 v / v).

36. A prebiotic fiber composition obtained from Pleurotus pulmonarius (PP), which comprises (1→3),(1→6) and (1→3)-β-D-glucan.70085-02 37. The prebiotic fiber composition of claim 36, which is lyophilized.

38. The prebiotic fiber composition of claim 36, which is insoluble or solubilized in water.

39. A method of promoting growth of Anaerostipes sp., Bacteroides uniformis, or both in the gut microbiome of an animal, which method comprises orally administering the prebiotic fiber composition of claim 30, alone or in further combination with live Anaerostipes sp., Bacteroides uniformis, or both, simultaneously or sequentially, whereupon growth of Anaerostipes sp., Bacteroides uniformis, or both in the gut microbiome of the animal is promoted.

40. A prebiotic fiber composition comprising: (a) a prebiotic fiber composition obtained from Gloeoporus thelephoroides (GT), which comprises (1→3),(1→6) and (1→3)-β-D-glucan; (b) a prebiotic fiber composition obtained from Ganoderma lucidum (GL), which comprises (1→3),(1→6) and (1→3)-β-D-glucan; (c) a prebiotic fiber composition obtained from Pholiota nameko (PN), which comprises (1→3),(1→6) and (1→3)-β-D-glucan; (d) a prebiotic fiber composition obtained from Pleurotus pulmonarius (PP), which comprises (1→3),(1→6) and (1→3)-β-D-glucan; (e) (a) and (b); (f) (a) and (c); (g) (a) and (d); (h) (b) and (c); (i) (b) and (d); (j) (c) and (d); or (k) (a), (b), (c), and (d).

41. The prebiotic fiber composition of claim 40, which is lyophilized.70085-02 42. The prebiotic fiber composition of claim 40, which is insoluble or solubilized in water.

43. A method of promoting growth of Anaerostipes sp., Bacteroides uniformis, or both in the gut microbiome of an animal, which method comprises orally administering the prebiotic fiber composition of claim 40, alone or in further combination with live Anaerostipes sp., Bacteroides uniformis, or both, simultaneously or sequentially, whereupon growth of Anaerostipes sp., Bacteroides uniformis, or both in the gut microbiome of the animal is promoted.