Abstract
Background
Dietary β-glucans from diverse sources exhibit varying prebiotic potentials, yet their comparative impacts on gut microbiota composition and short-chain fatty acid (SCFAs) production remain underexplored.
Aims
This study investigated the effects of three β-glucans—oat, mushroom (Lentinula edodes), and curdlan—on gut microbiota modulation, and SCFA profiles in mice.
Methods
Forty C57BL/6J mice were fed a low-fat diet supplemented with different β-glucans for 15 weeks. Gut microbiota composition was analyzed via 16S rRNA sequencing, and SCFAs levels were measured.
Results
Gut microbiota analysis revealed that oat β-glucan significantly reduced alpha diversity indices (observed species, ACE, and phylogenetic diversity) while increasing beneficial genera such as Prevotellaceae_UCG-001, Ruminiclostridium_5, Butyricicoccus, Ruminiclostridium_6, and Prevotellaceae_NK3B31_group. Oat β-glucan also elevated serum acetate, propionate, and lactate levels, whereas mushroom β-glucan selectively increased butyrate, and curdlan had no impact on SCFAs. Correlation analysis linked these SCFA enhancements to the enrichment of SCFA-producing bacteria (e.g., Ruminococcaceae, Lachnospiraceae).
Conclusions
These findings highlight the source-dependent bioactivity of β-glucans, positioning oat β-glucan as a potent modulator of the gut-brain axis through microbiota-SCFA interactions, with implications for dietary interventions targeting metabolic and cognitive health.
Supplementary Information
The online version contains supplementary material available at 10.1186/s40795-025-01160-9.
Keywords: Mice, Prebiotics, Gut microbiota, Β-gulcan, SCFAs
Introduction
The gut microbiota, a complex microbial community residing in the gastrointestinal tract, plays a pivotal role in host physiology through dynamic interactions with dietary components [1]. Among the metabolites produced by gut microbes, short-chain fatty acids (SCFAs), such as acetate, propionate, and butyrate, are critical mediators of these interactions. SCFAs are generated via the fermentation of dietary fibers and have been implicated in anti-inflammatory, immunomodulatory, and neuroprotective effects, linking gut microbial activity to systemic health [2, 3]. Consequently, dietary interventions aimed at modulating the gut microbiota and enhancing SCFA production have garnered significant attention as potential strategies to mitigate metabolic and neurodegenerative disorders [4].
β-glucans, a class of polysaccharides found in cereals, fungi, and microbial sources, are well-characterized dietary fibers known for their prebiotic properties [5]. These compounds resist digestion in the upper gastrointestinal tract and serve as substrates for microbial fermentation in the colon, thereby influencing microbial composition and metabolic output [5]. Emerging evidence suggests that the gut microbiota’s response to β-glucans may depend on their solubility, molecular weight, and branching patterns, which differ across sources [6]. Most existing studies focus on single-source β-glucans, systematic comparisons of β-glucans from distinct origins remain limited [7]. This knowledge gap hinders the rational selection of β-glucan sources for targeted dietary interventions [8]. Our study fills this gap by providing a comprehensive, head-to-head comparison of three structurally distinct β-glucans within the same experimental model, over a long-term intervention period. In this study, we aimed to (1) directly compare the prebiotic efficacy of oat, mushroom, and curdlan β-glucans in reshaping gut microbial diversity and composition; (2) evaluate their differential capacity to enhance systemic levels of key SCFAs and lactate; and (3) explore correlations between specific microbiota shifts and SCFA production. Our findings provide critical insights into the source-dependent bioactivity of β-glucans, advancing their potential application in personalized nutrition and gut-brain axis modulation.
Methods
Animals and treatment
Forty nine-week-old male C57BL/6J mice were purchased from the Experimental Animal Center of Xuzhou Medical University (Xuzhou, China, SCXK (Su) 2015-0009). This research was designed according to the National Institutes of Health (NIH) Guide for the Care and Use of Laboratory Animals and approved by the Ethics Committee of Laboratory Animals of Xuzhou Medical University (202007A192). Following habituation to the laboratory environment, mice were randomly assigned to four dietary groups (n = 10 per group): (1) Control group mice were fed a control diet; (2) Oat group mice were fed a control diet supplemented with β-glucan from oat bran, (3) Mushroom group mice were fed a control diet supplemented with β-glucan extracted from shiitake mushroom, and (4) Curdlan group mice were fed a control diet supplemented with curdlan-derived β-glucan. The composition of all diets matched that described in previous reports [9]. After 15 weeks food intake, all mice were euthanized using carbon dioxide (CO₂) following the AVMA Guidelines for the Euthanasia of Animals. The CO₂ was introduced into the cage at a flow rate of 50% cage volume displacement per minute (VDR/min), controlled by a flow meter. The CO₂ flow continued for 2 min (active exposure phase) to ensure complete displacement of the cage gas. The CO₂ was then turned off, and the cage remained undisturbed for the assigned passive exposure period. This flow rate (50% VDR/min) and exposure duration were selected based on prior laboratory data confirming that this exposure time resulted in cessation of respiration [10, 11]. Blood serum and the fresh cercal contents of mice were collected and stored in − 80 °C for further analyses.
Gut microbiota analysis
The microbiomes in cecal contents of mice were analyzed with 16S ribosomal RNA (rRNA) gene sequencing at the Illumina sequencing platform by Genedenovo Biotechnology Co., Ltd. (Guangzhou, China). The 16S rRNA gene sequencing was performed by the protocols described previously [9, 12]. The diversity analysis and the linear discriminant analysis coupled with effect size (LEfSe) were performed using the OmicShare tools, a free online platform for data analysis (http://www.omicshare.com/tools).
Measurement of short-chain fatty acids (SCFAs) and lactic acid
Freshly serum samples from the mice were collected to determine the SCFAs concentrations with gas chromatographymass spectrometry (GC-MS). The GC-MS analysis was performed by the protocols described previously [13, 14]. Briefly, this experiment was performed on an Agilent HP-INNOWAX capillary column. The temperature was set following the details described previously [14]. Helium was used as the carrier gas with a flow rate of 1.0 mL/min. The temperatures of the inlet, interface, and ionization source were 250 °C, 230 °C, and 250 °C, respectively. The lactic acid was sent to clinical laboratory of Affiliated Hospital of Xuzhou Medical University for determination.
Statistical analysis
The data are presented as means ± SEM. Statistical analysis was performed using the one-way analysis of variance (ANOVA) by SPSS (version 20, IBM Corporation, Chicago, IL, United States), followed by the post hoc Tukey test for comparisons among the groups [15, 16]. A P value < 0.05 was considered as statistically significant.
Results
Dietary β-glucans supplementation could regulate gut microbiota diversity of mice
Alpha diversity and beta diversity were performed to characterize microbial diversity after different β-glucans treatment. Alpha diversity metrics revealed reductions in observed species (F(3,20) = 8.745, P = 0.0007, Fig. 1A), ACE index (F(3,20) = 11.24, P = 0.0002, Fig. 1B), and phylogenetic diversity (F(3,20) = 10.35, P = 0.0003, Fig. 1C) across all β-glucan groups compared to control. Specifically, oat and curdlan supplementation significantly decreased observed species and ACE index, while oat uniquely reduced phylogenetic diversity (all P < 0.05, Fig. 1A-C). Beta diversity analysis (Principal coordinate analysis (PCoA) based on weighted UniFrac distances) demonstrated distinct clustering of the oat group, separating it from control, mushroom, and curdlan groups (Fig. 1D). Collectively, these findings indicate that oat β-glucan exerts the most pronounced modulatory effects on gut microbiota diversity among the tested β-glucans.
Fig. 1.
Dietary β-glucans supplementation could regulate gut microbiota diversity of mice. (A) Oat and Curdlan groups significantly decreased the observed species compared to control group. (B) Oat and Curdlan groups significantly decreased ACE index compared to control group. (C) Oat group significantly decreased the phylogenetic diversity compared to control group. (D) Principal coordinate analysis (PCoA) based on weighted UniFrac distances in the four groups. Data was presented as mean ± SEM. n = 6. *p < 0.05, ***p < 0.001.
Dietary β-glucans supplementation could regulate gut microbiota composition of mice
Linear discriminant analysis (LDA) effect size (LEfSe) revealed that oat supplementation significantly enriched Prevotellaceae_UCG-001, Ruminiclostridium_5, Butyricicoccus, Ruminiclostridium_6, and Prevotellaceae_NK3B31_group (LDA score > 2.0; Fig. 2A). Notably, these discriminative taxa belong primarily to the Prevotellaceae and Ruminococcaceae families. Conversely, mushroom selectively increased Ruminococcaceae_UCG_010 (Ruminococcaceae) and Enterorhabdus (Coriobacteriaceae) (LDA score > 2.0; Fig. 2B). Curdlan supplementation showed no significant genus-level alterations. These results demonstrate that β-glucan-mediated microbiota modulation is source-dependent, with oat exhibiting the broadest compositional impact.
Fig. 2.
Dietary β-glucans supplementation could regulate gut microbiota composition of mice. (A) Linear discriminant analysis (LDA) effect size (LEfSe) showing the most significantly abundant taxa enriched in microbiome from the oat group compared to the control group. (B) Linear discriminant analysis (LDA) effect size (LEfSe) showing the most significantly abundant taxa enriched in microbiome from the mushroom group compared to the control group.
Various β-glucans have different effects in increasing the short chain fatty acids and other organic acid
While dietary fibers are known to enhance SCFA production [5], this study provides the first comparative analysis of three distinct β-glucans on systemic SCFA profiles. Following 15-week supplementation, oat significantly elevated plasma acetate (F(3,30) = 5.347, P = 0.0045, Fig. 3A), propionate (F(3,31) = 16.22, P < 0.0001, Fig. 3B), and lactate (F(3,32) = 2.663, P = 0.0646, Fig. 3D) concentrations. In contrast, mushroom selectively increased butyrate levels (F(3,33) = 3.705, P = 0.021, Fig. 3C), while curdlan supplementation showed no significant effects on any SCFAs or lactate. These results demonstrate that oat β-glucan elicits the most comprehensive stimulation of microbial metabolites, affecting multiple SCFA production.
Fig. 3.
Various β-glucans have different effects in increasing the short-chain fatty acids and other organic acid. (A) Oat group significantly increased the concentrations of acetate acid in the serum of mice compared to control group. (B) Oat group significantly increased the concentrations of propionate acid in the serum of mice compared to control group. (C) Mushroom group significantly increased the concentrations of butyrate acid in the serum of mice compared to control group. (D) Oat group significantly increased the concentrations of lactic acid in the serum of mice compared to control group. Data was presented as mean ± SEM. n = 8–10. *p < 0.05, **p < 0.01, ***p < 0.001.
The correlation between short-chain fatty acid and bacterium
Since the SCFA are the major products from the microbial fermentative activity in the gut [17], so the correlation between SCFA and bacterium was analyzed. The level of acetate, propionate, butyrate and lactate were positively correlated with the most differentiating bacterial taxa, such as Butyricicoccus, Ruminococcaceae_NK4A214_group, Ruminococcaceae_UCG-010, Ruminiclostridium_5, Ruminiclostridium_6 (these five all belonging to Ruminococcaceae family); Bifdobacterium (Bifidobacteriaceae).
Besides, Lachnoclostridium, Lachnospiraceae_NK4A136_group, Lachnospiraceae_UCG-001, and Coprococcus_1, (these four all belonging to Lachnospiraceae family), Prevotellaceae_UCG-001, Prevotellaceae_NK3B31_group and Paraprevotella (these three belonging to Prevotellaceae family), were also positively correlated with SCFA productions (Fig. 4). However, the level of SCFA was negatively correlated with Oscillibactiter, Anaerotruncus and Odoribacter. These findings indicate that the elevated SCFA levels are likely mediated by β-glucan-induced alterations in the gut microbiota.
Fig. 4.
The correlation between short-chain fatty acids and bacterium. (A) The heatmap showing the Spearman’s rank correlation between the selected taxa of the microbiome and acetate, propionate, butyrate and lactate. P < 0.05 for significant correlations, R is noted.
Discussion
Our findings demonstrate a clear hierarchy among the tested β-glucans: oat β-glucan exhibited the most pronounced effects, mushroom (Lentinula edodes) β-glucan had selective impacts, and curdlan β-glucan showed negligible activity under the conditions tested.
Oat β-glucan significantly reduced alpha diversity—a state often associated with disease—this reduction was beneficial, as it reflected a selective enrichment of SCFA-producing bacteria. Genera significantly increased by oat β-glucan including Prevotellaceae_UCG-001, Ruminiclostridium_5, Butyricicoccus, Ruminiclostridium_6, Prevotellaceae_NK3B31_group, are strongly associated with fiber fermentation and SCFA-producing [17–21]. This suggests oat β-glucan promotes a more specialized, functionally efficient microbial community geared towards carbohydrate metabolism, rather than inducing pathological dysbiosis. The clear separation of the oat group in PCoA analysis further underscores its unique impact on overall microbial community. In contrast, the minimal effects of mushroom and curdlan β-glucans on microbial diversity and composition highlight the source-dependency of β-glucan bioactivity. Therefore, oat β-glucan is a potent, source-dependent prebiotic that uniquely shapes the gut microbiota into a beneficial composition.
At the genus level, oat significantly increased the Butyricicoccus, a gut SCFA-producing bacterium [20]. Oat β-glucan supplementation also significantly increased the relative abundance of Prevotellaceae_UCG-001 (family Prevotellaceae), Ruminiclostridium_5, and Ruminiclostridium_6 (family Ruminococcaceae). This finding is particularly noteworthy in light of a fecal microbiota transplantation (FMT) study demonstrating that aged donors—characterized by reduced abundance of Prevotellaceae and Ruminococcaceae—impaired spatial learning and memory in young recipient mice [22]. The observed elevation of these taxa in our oat-supplemented group suggests a potential shift toward a more youthful microbial profile. Conversely, mushroom β-glucan supplementation selectively increased Ruminococcaceae_UCG_010 (a documented butyrate-producer) [23] and Enterorhabdus. The enrichment of Ruminococcaceae_UCG_010 aligns mechanistically with the significant butyrate elevation observed in the mushroom group [24]. These findings position β-glucan as a potent dietary modulator of the gut environment.
The differential effects on SCFA profiles reinforce this source-specificity. Oat β-glucan significantly elevated systemic levels of acetate, propionate, and lactate. Acetate and propionate are key energy substrates and signaling molecules influencing host metabolism and immunity systemically [2, 3]. Lactate, while often an intermediate, can also influence immune function and serves as a precursor for SCFAs production [17]. The selective increase in butyrate by mushroom β-glucan is noteworthy, given butyrate’s paramount importance as the primary energy source for colonocytes and its potent anti-inflammatory and epigenetic regulatory roles [25]. The absence of significant SCFA changes with curdlan supplementation aligns with its lack of impact on microbiota composition. The correlation analysis robustly linked the enrichment of specific taxa to increased SCFA levels, providing mechanistic support for the observed metabolic shifts. The detection of elevated SCFAs and lactate in the systemic circulation underscores their potential to exert effects beyond the gut, including potential modulation of the gut-brain axis [26].
The superior efficacy of oat β-glucan can likely be attributed to its physicochemical properties—primarily its high solubility and specific molecular structure (mixed β-(1→3)/β-(1→4) linkages). These features enhance its accessibility to a broader range of fermentative bacteria in the proximal colon [5]. Mushroom β-glucans have short β (1, 6)-linked branches from a β (1, 3) backbone [5]. This structure confers potent immunomodulatory properties via direct interactions with immune cells, it can act as pathogen associated molecular pattern and bind to various pattern recognition receptors expressed on surface of immune cells thereby facilitating their activation and crosstalk [27]. It may render them less readily fermentable by a wide consortium of gut bacteria, potentially explaining the more limited (though significant for butyrate) microbial impact observed. Curdlan, a linear β-(1→3)-glucan, is highly insoluble and known for its gelling properties rather than fermentability [28], accounting for its lack of prebiotic effect in this model.
Limitations and future research recommendations
This study has several limitations. First, the functional potential for SCFA production was inferred from microbial taxonomy and correlation analyses rather than through direct measurement. Second, the study did not assess downstream physiological effects—such as gut barrier function, systemic inflammation, metabolic markers, or cognitive outcomes—which are necessary to confirm a functional impact on the gut-brain axis or overall metabolic health. Most importantly, the use of animal models limits the direct translation of results to humans due to interspecies biological differences.
To establish causality, future work should employ gnotobiotic or antibiotic-treated animal models colonized with specific microbial taxa. This approach can directly test their role in boosting SCFA production. Subsequent analyses must measure key downstream physiological outcomes—including gut barrier function, systemic inflammation, metabolic markers, and cognitive/behavioral endpoints—to substantiate claims of gut-brain axis mediation. Ultimately, these findings must be validated in humans through randomized controlled trials (RCTs) that account for inter-individual microbiome variability.
Conclusion
Our findings demonstrate that β-glucan prebiotic effects are highly source-dependent. Oat β-glucan was most effective, significantly increasing SCFA producers and elevating acetate, propionate, and lactate levels. Mushroom β-glucan selectively enhanced butyrate, while curdlan showed no significant activity. These findings highlight that not all β-glucans are equal, emphasizing the critical importance of source selection for dietary interventions targeting the gut microbiome, SCFA benefits, and even gut-brain axis health.
Supplementary Information
Below is the link to the electronic supplementary material.
Acknowledgements
We would like to acknowledge Professor Yinghua Yu’s group in Xuzhou Medical University for their support during conducting this research. We would also like to express gratitude for study participants, data collectors, friends and supervisors for their full support during the whole process of the study.
Abbreviations
- 16S rRNA
16S ribosomal RNA
- ACE
Abundance-based Coverage Estimator
- GC-MS
Gas chromatographymass spectrometry
- LDA
Linear discriminant analysis
- PCoA
Principal coordinate analysis
- SCFA
Short-chain fatty acid
- VDR
Volume displacement rate
Author contributions
M H designed research; R W, X Zh, H Q, X X, F Y performed research; M H, R W, X Zh, H Q, X X, Y Zh, R Zh, X L, Y Zh analyzed data; and M H, R W, X Zh, H Q wrote the paper, All authors reviewed the manuscript.
Funding
This work was supported by the Jiangsu Training Program of Innovation and Entrepreneurship for Undergraduates (202310313051Z, 202410313058Y, X202510313021); the National Demonstration Center for Experimental Basic Medical Science Education (Xuzhou Medical University) Student Science and Technology Innovation Project (2024BMS18), the Starting Foundation for Talents of Xuzhou Medical University (D2017007).
Data availability
All 16S rRNA raw data were submitted to the NCBI Sequence Read Archive (SRA) Database with the accession number: PRJNA801759.
Declarations
Ethics approval and consent to participate
This research was designed according to the National Institutes of Health (NIH) Guide for the Care and Use of Laboratory Animals and approved by the Ethics Committee of Laboratory Animals of Xuzhou Medical University. All the mice used for experiments were approved by the Institutional Animal Care Committee of Xuzhou Medical University following the Chinese Council on Animal Care Guidelines.
Consent for publication
Not applicable.
Competing interests
The authors declare no competing interests.
Footnotes
Publisher’s note
Springer Nature remains neutral with regard to jurisdictional claims in published maps and institutional affiliations.
Ruicong Wu, Xinyou Zhang and Haoyu Qin contributed equally to this work.
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Associated Data
This section collects any data citations, data availability statements, or supplementary materials included in this article.
Data Citations
Supplementary Materials
Data Availability Statement
All 16S rRNA raw data were submitted to the NCBI Sequence Read Archive (SRA) Database with the accession number: PRJNA801759.