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29 pages, 5748 KB  
Article
Metatranscriptome Analysis of Sheep Rumen Reveals Methane Production Changes Induced by Moringa oleifera as a Dietary Supplement
by Alicia Alejandra Grijalva-Hinojos, Vicente Arnau, Wladimiro Díaz, Samuel Piquer, Daniel Díaz-Plascencia, Yamicela Castillo-Castillo, Joel Domínguez-Viveros and Perla Lucia Ordoñez-Baquera
Fermentation 2025, 11(10), 568; https://doi.org/10.3390/fermentation11100568 - 1 Oct 2025
Abstract
Global warming has become a significant public health concern, with intensive livestock farming as a major contributor. To mitigate greenhouse gas emissions, strategies such as manipulating the ruminal environment with dietary additives are essential. This study evaluated Moringa oleifera, a globally widespread [...] Read more.
Global warming has become a significant public health concern, with intensive livestock farming as a major contributor. To mitigate greenhouse gas emissions, strategies such as manipulating the ruminal environment with dietary additives are essential. This study evaluated Moringa oleifera, a globally widespread tree with antioxidant, multivitamin, protein-rich, and anti-inflammatory properties, as a feed additive. Rumen fluid was collected from three Pelibuey sheep, homogenized, and subjected to an in vitro fermentation study for 48 h with three alfalfa/moringa ratio treatments: T0 Control (100:0), T1 Low (85:15), and T2 High (70:30). Total RNA was extracted, followed by high-definition sequencing of the metatranscriptome. The sequencing yielded approximately 456 million sequences. A total of 117 phyla were identified and approximately 1300 genera were mapped. Predominant phylum differed by treatment: T0, Firmicutes; T1, Proteobacteria; and T2 with Synergistetes, at least one sample per treatment. Archaea were nearly absent in T1, which explains a statistically significant decrease in methane production. In the Gene Set Enrichment Analysis (GSEA), it was observed that one of the metabolic pathways with a statistically significant difference (p-value < 0.05) was that of methane, specifically in the low moringa treatment (T1) compared to the control (T0). From the functional analysis, differentially expressed enzymes were identified, some of which are involved in the methane metabolic pathway, such as formate dehydrogenase (EC 1.17.1.9) and glycine hydroxymethyltransferase (EC 2.1.2.1), which are intermediates in methane formation. These results suggest that 15% Moringa oleifera supplementation alters ruminal microbiota, reduces archaeal activity, and suppresses methane-related pathways. These findings provide molecular evidence supporting the potential of M. oleifera as a methane mitigation strategy in ruminant nutrition. Full article
(This article belongs to the Special Issue Ruminal Fermentation: 2nd Edition)
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24 pages, 1804 KB  
Review
Digestive Neurobiology in Autism: From Enteric and Central Nervous System Interactions to Shared Genetic Pathways
by Raz Robas, Utkarsh Tripathi, Wote Amelo Rike, Omveer Sharma and Shani Stern
Int. J. Mol. Sci. 2025, 26(19), 9580; https://doi.org/10.3390/ijms26199580 - 1 Oct 2025
Abstract
Social communication difficulties characterize autism spectrum disorders (ASD). Gastrointestinal (GI) symptoms are more common in ASD than in the general population. The identification of GI problems in individuals with ASD is challenging due to their altered pain perception and irregular behaviors. Importantly, GI [...] Read more.
Social communication difficulties characterize autism spectrum disorders (ASD). Gastrointestinal (GI) symptoms are more common in ASD than in the general population. The identification of GI problems in individuals with ASD is challenging due to their altered pain perception and irregular behaviors. Importantly, GI symptoms and ASD can potentially aggravate each other. However, it is unclear if GI problems cause ASD symptoms or vice versa. A crosstalk between the digestive system, gut microbiota, and the central and enteric nervous systems (CNS and ENS, respectively) has been repeatedly reported. The ENS regulates the GI tract with the CNS and the autonomic nervous system (ANS), as well as independently through specific neural circuits. Several mechanisms contribute to GI problems in ASD, including genetic mutations that affect the ENS, dysregulation of the ANS, alterations in gut microbiota, unhealthy dietary preferences, and changes in metabolomic profiles. Furthermore, studies have shown molecular and cellular differences in the GI biopsy of children with and without ASD. These findings highlight the unique nature of GI issues in ASD, underscoring the importance of further investigating the changes that occur in the digestive system and ENS in ASD models. Full article
(This article belongs to the Section Molecular Neurobiology)
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21 pages, 1128 KB  
Article
Methods of Phytic Acid Reduction in Bitter Lupine Seeds and Their Effects on the Microbiota of Calves
by Barbara Płacheta-Kwiatkowska, Oliwia Brodowicz, Weronika Cieciura-Włoch, Małgorzata Wlaźlak, Agnieszka Wilkowska, Ilona Motyl and Joanna Berłowska
Agriculture 2025, 15(19), 2061; https://doi.org/10.3390/agriculture15192061 - 30 Sep 2025
Abstract
Phytic acid is an antinutritional factor present in lupine seeds, which limits the bioavailability of essential minerals such as calcium, iron, and zinc. This study evaluated different methods of reducing phytic acid in bitter lupine (Lupinus angustifolius) and investigated the effects [...] Read more.
Phytic acid is an antinutritional factor present in lupine seeds, which limits the bioavailability of essential minerals such as calcium, iron, and zinc. This study evaluated different methods of reducing phytic acid in bitter lupine (Lupinus angustifolius) and investigated the effects of the resulting reduction in phytic acid on the composition of gut microbiota. Bitter lupine is a legume rich in protein and fiber, but its high phytic acid content can limit mineral bioavailability. Four processing methods were compared as follows: thermal treatment, enzymatic hydrolysis with phytase, spontaneous fermentation, and controlled fermentation using lactic acid bacteria. Controlled fermentation resulted in the highest phytic acid reduction (96.37%), significantly improving mineral availability. Simulated digestion revealed that the fermented lupine feed positively influenced gut microbiota, increasing Lactobacillus abundance. Enzymatic and thermal treatments preserved more protein. However, they were less effective at removing phytic acid. These findings highlight controlled fermentation as a promising strategy for improving the nutritional value of lupine-based feed, offering a sustainable alternative to soybean-based livestock diets. Full article
(This article belongs to the Section Farm Animal Production)
18 pages, 716 KB  
Communication
Significant Association Between Abundance of Gut Microbiota and Plasma Levels of microRNAs in Individuals with Metabolic Syndrome and Their Potential as Biomarkers for Metabolic Syndrome: A Pilot Study
by Sanghoo Lee, Jeonghoon Hong, Yiseul Kim, Hee-Ji Choi, Jinhee Park, Jihye Yun, Yun-Tae Kim, Kyeonghwan Choi, SaeYun Baik, Mi-Kyeong Lee and Kyoung-Ryul Lee
Genes 2025, 16(10), 1161; https://doi.org/10.3390/genes16101161 - 30 Sep 2025
Abstract
Background/Objectives: The relationship between gut microbiota (GM) and microRNAs (miRs) related to lipid metabolism in individuals with metabolic syndrome (MetS) remains unclear. This pilot study examined the relationship between Bacteroidetes and Firmicutes abundance at the phylum level and the plasma levels of miR-122 [...] Read more.
Background/Objectives: The relationship between gut microbiota (GM) and microRNAs (miRs) related to lipid metabolism in individuals with metabolic syndrome (MetS) remains unclear. This pilot study examined the relationship between Bacteroidetes and Firmicutes abundance at the phylum level and the plasma levels of miR-122 and miR-370, both of which are associated with lipid metabolism, in Korean individuals with MetS and in healthy controls. We also evaluated the potential of these miRs as biomarkers for MetS. Methods: This study enrolled 7 individuals with MetS and 8 controls. The abundance of GM was analyzed by 16S rRNA amplicon sequencing. To evaluate the relationship between the dominant phyla in the 2 groups, the log ratio of Firmicutes to Bacteroidetes (F/B) was calculated using a centered log-ratio (CLR) transformation. The abundance of the 2 plasma miRs was also quantified by real-time quantitative PCR (RT-qPCR). Pearson’s and Spearman’s correlation analyses were then performed to evaluate the relationship between Bacteroidetes and Firmicutes abundance, the clinical parameters, and plasma levels of the 2 miRs. Additionally, the area under the curve (AUC) value of the receiver operating characteristic (ROC) curve was calculated to evaluate the potential of the 2 miRs as MetS biomarkers. Results: The 2 most abundant phyla were Bacteroidetes and Firmicutes. Bacteroidetes made up an average of 24.7% in the MetS group and 69.7% in the control group. Meanwhile, the average abundance of Firmicutes was 69.8% in the MetS group and 26.5% in the control group. The log F/B ratios in the MetS and control groups were 0.7 ± 0.5 and −0.4 ± 0.1 (p < 0.001), respectively. FDR analysis revealed significant correlations between Bacteroidetes abundance and BMI, DBP, FBG, total chol, insulin and HOMA-IR (FDR-adjusted p < 0.05), as well as between Firmicutes abundance and BMI, FBG, total chol, insulin and HOMA-IR (FDR-adjusted p < 0.05). Plasma levels of the 2 miRs differed significantly between the MetS and control groups: miR-122 (1.43 vs. 0.73; p = 0.0065) and miR-370 (1.39 vs. 0.83; p = 0.0089). The AUC values for miR-122 and miR-370 were 0.946 (p < 0.001) and 0.964 (p < 0.001), respectively. Pearson’s and Spearman’s correlation analyses revealed significant negative correlations between Bacteroidetes abundance and levels of miR-122 (p = 0.0048 and p = 0.0045, respectively) and miR-370 (p = 0.0003 and p < 0.0001, respectively), as well as significant positive correlations between Firmicutes abundance and levels of miR-122 (p = 0.0038 and p = 0.0027, respectively) and miR-370 (p = 0.0004 and p < 0.0001, respectively). However, as our exploratory findings were based on a small sample size, the high correlation results may partly reflect the separation between the MetS and control groups. Conclusions: Our exploratory findings suggest that the GM abundances of individuals with MetS may be significantly associated with plasma levels of miR-122 and miR-370, which are related to lipid metabolism. These miRs may therefore serve as potential MetS biomarkers. Full article
(This article belongs to the Section RNA)
22 pages, 4897 KB  
Article
Fabrication of Next-Generation Skin Scaffolds: Integrating Human Dermal Extracellular Matrix and Microbiota-Derived Postbiotics via 3D Bioprinting
by Sultan Golpek Aymelek, Billur Sezgin Kizilok, Ahmet Ceylan and Fadime Kiran
Polymers 2025, 17(19), 2647; https://doi.org/10.3390/polym17192647 - 30 Sep 2025
Abstract
This study presents the development of an advanced three-dimensional (3D) bioprinted skin scaffold integrating sodium alginate (SA), gelatin (Gel), human skin-derived decellularized extracellular matrix (dECM), and microbiota-derived postbiotics. To ensure a biocompatible and functional ECM source, human skin samples collected during elective aesthetic [...] Read more.
This study presents the development of an advanced three-dimensional (3D) bioprinted skin scaffold integrating sodium alginate (SA), gelatin (Gel), human skin-derived decellularized extracellular matrix (dECM), and microbiota-derived postbiotics. To ensure a biocompatible and functional ECM source, human skin samples collected during elective aesthetic surgical procedures were utilized. Following enzymatic treatment, the dermal layer was carefully separated from the epidermis and subjected to four different decellularization protocols. Among them, Protocol IV emerged as the most suitable, achieving significant DNA removal while maintaining the structural and biochemical integrity of the ECM, as confirmed by Fourier-transform infrared spectroscopy. Building on this optimized dECM-4, microbiota-derived postbiotics from Limosilactobacillus reuteri EIR/Spx-2 were incorporated to further enhance the scaffold’s bioactivity. Hybrid scaffolds were then fabricated using 7% Gel, 2% SA, 1% dECM-4, and 40 mg/mL postbiotics in five-layered grid structures via 3D bioprinting technology. Although this composition resulted in reduced mechanical strength, it exhibited improved hydrophilicity and biodegradability. Moreover, antimicrobial assays demonstrated inhibition zones of 16 mm and 13 mm against methicillin-resistant Staphylococcus aureus (MRSA, ATCC 43300) and Pseudomonas aeruginosa (ATCC 27853), respectively. Importantly, biocompatibility was confirmed through in vitro studies using human keratinocyte (HaCaT) cells, which adhered, proliferated, and maintained normal morphology over a 7-day culture period. Taken together, these findings suggest that the engineered hybrid scaffold provides both regenerative support and antimicrobial protection, making it a strong candidate for clinical applications, particularly in the management of chronic wounds. Full article
(This article belongs to the Special Issue Polymers for Aesthetic Purposes)
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18 pages, 463 KB  
Review
Enteral Nutrition in Pediatric Crohn’s Disease: New Perspectives
by Viviana Fara Brindicci, Rosangela Grieco, Roberta Giusy Ruiz, Sabrina Cardile, Teresa Capriati, Chiara Maria Trovato, Giulia Bolasco, Daniela Knafelz, Fiammetta Bracci, Arianna Alterio, Francesca Ferretti, Domenica Elia, Elena Spinetti, Ruggiero Francavilla and Antonella Diamanti
Nutrients 2025, 17(19), 3124; https://doi.org/10.3390/nu17193124 - 30 Sep 2025
Abstract
Background/Objectives: The efficacy of exclusive enteral nutrition (EEN) on the induction of remission of Crohn’s disease (CD) has been demonstrated with different diets (elemental, semi-elemental, and polymeric). A narrative review was conducted to assess the effects of different enteral diets in pediatric [...] Read more.
Background/Objectives: The efficacy of exclusive enteral nutrition (EEN) on the induction of remission of Crohn’s disease (CD) has been demonstrated with different diets (elemental, semi-elemental, and polymeric). A narrative review was conducted to assess the effects of different enteral diets in pediatric CD patients, considering the hypothesis that manipulating the nutritional key ingredients may enhance the clinical efficacy. Methods: An extensive literature search was performed across PubMed, Embase, and the Cochrane Library, covering all records published up to 27 July 2025. Both pediatric and adult studies were considered, and nutritional composition was compared with remission rates. Results: Twelve studies involving patients with active CD treated with EEN were found. Most studies were conducted with polymeric diets (n = 8), which achieved a high remission rate (up to 85%), thus confirming their advantage over other EEN diets. Conclusions: EEN with polymeric diets satisfies the need to revert the acute inflammation in most pediatric CD patients. Polymeric formulas have two advantages: (a) they contain transforming growth factor-β (TGF-β), which exerts anti-inflammatory effects on intestinal epithelial cells, and (b) they have a mixed-fat composition, including saturated fatty acids (SFAs), monounsaturated fatty acids (MUFAs), polyunsaturated fatty acids (PUFAs) as well medium-chain triglycerides (MCTs), which provides better results than EEN diets enriched with single-fat components. However, pathophysiological evidence shows gut microbiota alterations after EEN begins, despite clinical improvement. So, a potential strategy to enhance the efficacy of polymeric diets may be fiber enrichment. Full article
(This article belongs to the Special Issue Dietary and Nutritional Therapies to Improve Digestive Disorders)
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15 pages, 944 KB  
Review
TMAO and Cardiovascular Disease: Exploring Its Potential as a Biomarker
by Octavian Amaritei, Oana Laura Mierlan, Ciprian Adrian Dinu, Iulia Chiscop, Madalina Nicoleta Matei, Cristian Gutu and Gabriela Gurau
Medicina 2025, 61(10), 1767; https://doi.org/10.3390/medicina61101767 - 30 Sep 2025
Abstract
Gut microbiota has increasingly been shown to exert effects beyond the gastrointestinal tract, some of which are mediated through its metabolites, such as trimethylamine N-oxide (TMAO)—a compound converted by gut bacteria from dietary choline found predominantly in animal products that is associated with [...] Read more.
Gut microbiota has increasingly been shown to exert effects beyond the gastrointestinal tract, some of which are mediated through its metabolites, such as trimethylamine N-oxide (TMAO)—a compound converted by gut bacteria from dietary choline found predominantly in animal products that is associated with cardiovascular disease (CVD). However, a significant gap persists in human clinical trials assessing its potential causal role. This narrative review aims to present the current understanding of the gut microbiome, TMAO, and their relationship with CVD, while proposing future directions that may support the use of TMAO as a biomarker and guide potential interventions to reduce its harmful impact. Both animal and human studies have demonstrated a link between TMAO and CVD, with animal studies also indicating a causal effect—showing increased cardiovascular risk following TMAO administration and reduced risk when TMAO is eliminated. While direct extrapolation from animal models to humans is limited due to biological differences, these findings offer a foundation for the development of well-designed clinical trials in human populations. Although direct approaches to target TMAO—such as trimethylamine (TMA) lyase inhibitors and antisense oligonucleotide (ASO) therapy—have shown promising results in animal studies, they have yet to be investigated in human trials, leaving indirect strategies such as dietary changes and probiotics as the only currently available options. Full article
(This article belongs to the Section Cardiology)
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13 pages, 1942 KB  
Article
Characterization of Gut Bacteria in Natural Populations of Sand Flies (Diptera: Psychodidae) from Endemic and Non-Endemic Areas of Leishmaniasis in Morocco
by Mohamed Daoudi, Abdelkrim Outammassine, El Mahdi Redouane, Souad Loqman, Mohamed Hafidi, Ali Boumezzough, Martin Olivier, Samia Boussaa and Momar Ndao
Microorganisms 2025, 13(10), 2279; https://doi.org/10.3390/microorganisms13102279 - 30 Sep 2025
Abstract
Leishmaniasis is a vector-borne parasitic disease caused by Leishmania spp., transmitted to humans by phlebotomine sand flies. The development of Leishmania into infective metacyclic promastigotes occurs within the sand fly gut, where the bacterial microbiota plays a pivotal role in parasite development and [...] Read more.
Leishmaniasis is a vector-borne parasitic disease caused by Leishmania spp., transmitted to humans by phlebotomine sand flies. The development of Leishmania into infective metacyclic promastigotes occurs within the sand fly gut, where the bacterial microbiota plays a pivotal role in parasite development and transmission dynamics. This study aimed to characterize the gut bacterial composition of phlebotomine sand flies collected from both endemic (Lalla Aaziza) and non-endemic (Marrakech) regions of leishmaniasis in Morocco. We investigated the microbiota of Phlebotomus papatasi, P. sergenti, P. perniciosus, and P. longicuspis, all proven vectors of cutaneous and visceral leishmaniasis in the Old World, including Morocco, as well as Sergentomyia minuta, a potential vector in the Mediterranean basin. Gut bacteria were isolated using conventional microbiological techniques and identified by MALDI-TOF mass spectrometry. Fifteen bacterial strains from three phyla were identified, with Bacillus pumilus being the most frequently detected species. Significant differences in colony-forming unit (CFU) counts and bacterial richness were observed between sand fly species and collection sites. Notably, Bacillus simplex (in P. papatasi), Nocardia ignorata (in P. sergenti), and Serratia spp. (in P. longicuspis) were identified for the first time in these vectors. This study is the first to investigate the gut bacterial composition of sand flies in Morocco, revealing species and locality-dependent differences in microbial communities. The predominance of Bacillus spp., particularly B. pumilus, suggests a potentially influential role in sand fly physiology and vector competence. Furthermore, the novel detection of B. simplex, N. ignorata, and Serratia spp. underscores previously unrecognized microbial associations that warrant further investigation. These findings provide a critical baseline for future studies exploring the microbiota-mediated modulation of sand fly–Leishmania interactions. Full article
(This article belongs to the Section Medical Microbiology)
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24 pages, 7021 KB  
Article
Goblet Cells and Mucus Composition in Jejunum and Ileum Containing Peyer’s Patches and in Colon: A Study in Pigs
by Vladimir Ginoski, José Luis Cortés Sánchez, Stefan Kahlert, Johannes Schulze Holthausen, Łukasz Grześkowiak, Jürgen Zentek and Hermann-Josef Rothkötter
Animals 2025, 15(19), 2852; https://doi.org/10.3390/ani15192852 - 29 Sep 2025
Abstract
The intestinal mucus layer is a dynamic protective barrier that maintains gut homeostasis, supports immune defense, and regulates host–microbiota interactions. Rodent models have yielded valuable insights, but their intestinal structure and physiology differ from those of humans and pigs. By contrast, the omnivorous [...] Read more.
The intestinal mucus layer is a dynamic protective barrier that maintains gut homeostasis, supports immune defense, and regulates host–microbiota interactions. Rodent models have yielded valuable insights, but their intestinal structure and physiology differ from those of humans and pigs. By contrast, the omnivorous pig shares closer anatomical and immunological features with humans, making it a relevant large-animal model in translational studies. In this study, we established a histological workflow for porcine intestine by combining Carnoy’s fixation with Alcian Blue–Periodic Acid–Schiff and Mucicarmine staining. This enabled accurate visualization and quantification of goblet-cell density and mucus thickness across intestinal segments, with a particular focus on Peyer’s patches—key sites of immune surveillance. Both stains produced consistent results. We observed a clear proximal-to-distal gradient, from jejunum to colon, in mucus thickness: the colon displayed the thickest layer (~100 μm), whereas the follicle-associated epithelium over Peyer’s patches in the jejunum and ileum showed a markedly thinner layer (<12 μm) and fewer goblet cells. Immunofluorescence further revealed strong cytokeratin-18 expression in goblet cells, delineating their morphology and polarity. These findings demonstrate region-specific differences in mucus architecture and goblet-cell distribution that likely reflect specialized immune functions, advancing our understanding of the intestinal barrier and informing future strategies to support gut health and immunity. Full article
(This article belongs to the Section Pigs)
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22 pages, 4767 KB  
Article
Diversity and Function Potentials of Seed Endophytic Microbiota in a Chinese Medicinal Herb Panax notoginseng
by Hong-Yan Hu, Yun Wen, Shu-Cun Geng, Yu-Nuo Zhang, Yu-Bo Zhao, Xiao-Xia Pan, You-Yong Zhu, Xia-Hong He and Ming-Zhi Yang
Horticulturae 2025, 11(10), 1162; https://doi.org/10.3390/horticulturae11101162 - 29 Sep 2025
Abstract
As an important complementation of plant genetic traits, seed endophytes (SEs) have garnered significant attention due to their crucial roles in plant germination and early seedling establishment. In this study, we employed both culture-dependent and amplicon sequencing-based approaches to characterize the endophytic microbiome [...] Read more.
As an important complementation of plant genetic traits, seed endophytes (SEs) have garnered significant attention due to their crucial roles in plant germination and early seedling establishment. In this study, we employed both culture-dependent and amplicon sequencing-based approaches to characterize the endophytic microbiome in seed samples derived from different individual Panax notoginseng plants. Additionally, we evaluated the antagonistic activity of isolated culturable bacterial SEs against the root rot pathogens Fusarium solani and F. oxysporum. Our results demonstrated that a greater sampling quantity substantially increased the species richness (Observed OTUs) and diversity of seed endophytic microbiota, underscoring the importance of seed population size in facilitating the vertical transmission of diverse endophytes to progeny. The endophytic communities (including both fungi and bacteria) exhibited a conserved core microbiota alongside host-specific rare taxa, forming a phylogenetically and functionally diverse endophytic resource pool. Core bacterial genera included Streptococcus, Methylobacterium-Methylorubrum, Sphingomonas, Burkholderia-Caballeronia-Paraburkholderia, Pantoea, Halomonas, Acinetobacter, Pseudomonas, Vibrio, and Luteibacter, while core fungal genera comprised Davidiella, Thermomyces, Botryotinia, Myrothecium, Haematonectria, and Chaetomium. Among 256 isolated endophytic bacterial strains, 11 exhibited strong inhibitory effects on the mycelial growth of F. solani and F. oxysporum. Further evaluation revealed that two antagonistic strains, Bacillus cereus and B. toyonensis, significantly enhanced seed germination and plant growth in P. notoginseng, and effectively suppressed root rot disease in seedlings. These findings highlight the potential use of SEs as biocontrol agents and growth promoters in sustainable agriculture. Full article
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17 pages, 2087 KB  
Article
Integrated Analysis of Carotenoid Metabolism, Lipid Profiles, and Gut Microbiota Reveals Associations Fundamental to Skin Pigmentation in Lingshan Chickens
by Shengting Deng, Weiguang Yang, Shengdi Hu, Long Li, Jianhua He and Guozhi Bian
Animals 2025, 15(19), 2832; https://doi.org/10.3390/ani15192832 - 28 Sep 2025
Abstract
Skin color is a crucial phenotypic trait in poultry that influences consumer preference, market value, and breed identification. However, the mechanisms underlying skin color variation in Lingshan chickens remain poorly understood. This study aimed to elucidate the physiological, metabolic, and microbial characteristics associated [...] Read more.
Skin color is a crucial phenotypic trait in poultry that influences consumer preference, market value, and breed identification. However, the mechanisms underlying skin color variation in Lingshan chickens remain poorly understood. This study aimed to elucidate the physiological, metabolic, and microbial characteristics associated with skin color differences in male Lingshan chickens. A total of 210 castrated male Lingshan chickens were categorized into white-shanked (WS), yellow-shanked (YS), and red-shanked (RS) groups based on the Roche color fan scores. The results showed that chickens in the YS and RS groups exhibited significantly higher body weights and pigmentation levels in the shank, breast, and abdominal skin compared to those in the WS group (p < 0.05). Serum concentrations of triglycerides (TG), total cholesterol (TC), high-density lipoprotein (HDL), low-density lipoprotein (LDL), and very-low-density lipoprotein (VLDL) were markedly elevated in RS chickens. Additionally, carotenoid profiles revealed higher deposition of lutein and β-carotene in the skin and adipose tissues of YS and RS birds. Gene expression analysis indicated differential regulation of carotenoid transport and metabolism-related genes among groups. Furthermore, 16S rRNA sequencing of cecal microbiota revealed significant compositional shifts in microbial communities associated with shank pigmentation. Collectively, these findings suggest that differences in shank color in Lingshan chickens are closely linked to lipid metabolism, carotenoid transport, and gut microbiota composition. This study provides novel insights into the biological mechanisms driving skin pigmentation, offering valuable implications for breeding and functional trait selection in indigenous chicken populations. Full article
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18 pages, 1240 KB  
Article
Short-Chain Fatty Acid Profiles in Amyotrophic Lateral Sclerosis: Longitudinal Effects of Disease and Mediterranean Diet Intervention
by Anca Moțățăianu, Valentin Ion, Mihai Dumitreasă, Ioana Ormenișan, Lenard Farczadi, Sebastian Andone, Rodica Bălașa and Medeea Maria Roman
Biomolecules 2025, 15(10), 1380; https://doi.org/10.3390/biom15101380 - 28 Sep 2025
Abstract
Background: Amyotrophic lateral sclerosis (ALS) evolution is influenced by many dietary factors, biochemical and hormonal inter-relations and gut microbiota. This study focuses on dynamics by conducting a plasmatic quantitative analysis of six of the main short-chain fatty acids (SCFAs) for ALS patients [...] Read more.
Background: Amyotrophic lateral sclerosis (ALS) evolution is influenced by many dietary factors, biochemical and hormonal inter-relations and gut microbiota. This study focuses on dynamics by conducting a plasmatic quantitative analysis of six of the main short-chain fatty acids (SCFAs) for ALS patients and the shifts in circulating SCFA profiles during ALS progression as well as their potential responsiveness or change due to dietary modulation. Methods: A 12-month prospective study in parallel with control group determinations was conducted. The patients diagnosed with ALS were evaluated at the start of the study (T0) followed by a six-month observation time frame (T1) and after another six months of a Mediterranean diet intervention (T2). Plasma SCFAs were determined using liquid chromatography coupled to mass spectrometry to showcase the plasmatic profiles. Correlation between plasma levels of SCFAs and patients’ clinical characteristics next to correlations between plasma SCFA levels at T1 and T2 were performed. Results: A significant increase between control group and patients at T0 was observed for acetic, propionic, butyric and hydroxy-butyric acid. Hexanoic acid levels stagnated and 4-methyl-valeric acid concentrations decreased. Evolutions from T1 and T2 impacted acetate, propionate and 4-methyl-valerate. Conclusions: The study offers a better understanding regarding the differences in SCFA levels in ALS patients. The Mediterranean diet may impact the levels of acetic and propionic acid, indicating the modulation of SCFA production by gut microbiota. Full article
(This article belongs to the Section Bioinformatics and Systems Biology)
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20 pages, 3355 KB  
Article
The Impact of Weizmannia coagulans BC99 on Anxiety and Depression: An 8-Week Clinical Pilot Study Through the Gut Microbiota–Brain Axis
by Shanshan Tie, Yujia Pan, Chenguang Pang, Azeem Saman, Yao Dong, Shuguang Fang, Jianguo Zhu, Ying Wu and Shaobin Gu
Nutrients 2025, 17(19), 3087; https://doi.org/10.3390/nu17193087 - 28 Sep 2025
Abstract
Background: An imbalance in the immune system, stress response, and gut microbiota can contribute to the onset and progression of anxiety and depression. This pilot study aimed to explore the effect of Weizmannia coagulans BC99 on anxiety and depression through the gut [...] Read more.
Background: An imbalance in the immune system, stress response, and gut microbiota can contribute to the onset and progression of anxiety and depression. This pilot study aimed to explore the effect of Weizmannia coagulans BC99 on anxiety and depression through the gut microbiota–brain axis. Methods: A total of 79 participants with Hamilton Depression Rating Scale (HAMD-17) ≥ 8 or Hamilton Anxiety Rating Scale (HAMA-14) ≥ 7 were included and completed the study. The participants were randomly assigned to either the placebo group or the BC99 intervention group. The intervention lasted 8 weeks, with participants receiving either dextrin (3 g/day) or BC99 probiotics (3 g/day, 5 × 109 CFU) daily. Pre- and post-intervention comparisons were made on HAMD and HAMA scores, inflammatory cytokines, neurotransmitters, gut microbiota, and short-chain fatty acids. Results: Results showed that after 8 weeks, BC99 and placebo intervention were effective in reducing HAMD and HAMA scores, and HAMD and HAMA scores in BC99 group were reduced by 2.40 and 5.53 points compared the placebo group, and the response and remission rates were also higher than that of the placebo, but there was no significant difference. BC99 also could regulate the levels of inflammatory cytokines IL-17 and IL-10, and increase neurotransmitter levels of γ-GABA and NO. Moreover, compared with the placebo group, BC99 also enhanced the abundance of beneficial bacteria such as Faecalibactrium, Megamonas, Dialister, and Agathobacter, which were closely associated with clinical indicators of mental disorders, and increased the production of short-chain fatty acids. Conclusions: These preliminary findings suggested that BC99 might alleviate anxiety and depression symptoms by regulating the level of neurotransmitters or the change of microbiota, which needed further verification in subsequent animal or clinical experiments. Full article
(This article belongs to the Section Nutrition and Metabolism)
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9 pages, 1152 KB  
Article
Distinct Breast Tissue Microbiota Profiles in Early-Stage Breast Cancer: A Prospective Study in Turkish Women
by Mehmet Fatih Özsaray, Turgay Şimşek, Deniz Sünnetçi Akkoyunlu, Naci Çine and Nuh Zafer Cantürk
Life 2025, 15(10), 1518; https://doi.org/10.3390/life15101518 - 26 Sep 2025
Abstract
Background: This pilot study aimed to investigate the relationship between the breast tissue microbiota and breast cancer in Turkish women. We compared cancerous and adjacent normal breast tissues, as well as stool samples, obtained during breast-conserving surgery. Methods: In this prospective study, paired [...] Read more.
Background: This pilot study aimed to investigate the relationship between the breast tissue microbiota and breast cancer in Turkish women. We compared cancerous and adjacent normal breast tissues, as well as stool samples, obtained during breast-conserving surgery. Methods: In this prospective study, paired tumor and normal breast tissue samples, together with preoperative stool samples, were analyzed using 16S rRNA sequencing. Diversity indices and relative abundance differences were calculated, with effect sizes, 95% confidence intervals, and false discovery rate (FDR) corrections reported where appropriate. Results: A total of 22 patients with early-stage breast cancer were included (mean age 58.3 ± 12.7 years, mean BMI 28.9 ± 3.1 kg/m2). Distinct compositional shifts were observed between tumor and normal tissues, with Ruminococcus, Eubacterium, Actinobacteria (phylum), and Stenotrophomonas enriched in tumor samples, while Lactobacillus, Staphylococcus, Bifidobacterium, and Faecalibacterium were more abundant in normal tissues. No consistent associations were identified between fecal and breast tissue microbiota. Limitations: The small sample size, absence of healthy tissue or stool controls, and reliance on 16S rRNA sequencing limit the generalizability and functional interpretation of these findings. Conclusions: Despite these limitations, this study demonstrates localized microbial differences between tumor and adjacent normal breast tissues. Larger, multi-center studies with healthy controls and functional omics approaches are warranted to clarify the biological relevance and potential clinical implications. Full article
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Article
Infantile Anemia and Iron Treatments Affect the Gut Microbiome of Young Rhesus Monkeys
by Christopher L. Coe, Gabriele R. Lubach, Wellington Z. Amaral, Gregory J. Phillips, Mark Lyte, Michael K. Georgieff, Raghavendra B. Rao and James R. Connor
Microorganisms 2025, 13(10), 2256; https://doi.org/10.3390/microorganisms13102256 - 26 Sep 2025
Abstract
The influence of iron deficiency anemia and iron treatments on the gut microbiome was evaluated in young rhesus monkeys. First, the hindgut bacterial profiles of 12 iron-deficient anemic infants were compared to those of 9 iron-sufficient infants at 6 months of age, a [...] Read more.
The influence of iron deficiency anemia and iron treatments on the gut microbiome was evaluated in young rhesus monkeys. First, the hindgut bacterial profiles of 12 iron-deficient anemic infants were compared to those of 9 iron-sufficient infants at 6 months of age, a time when the risk of anemia is high due to rapid growth. After this screening, the anemic monkeys were treated with either parenteral or enteral iron. Seven monkeys were injected intramuscularly with iron dextran, the typical weekly treatment used in veterinary practice. Four other anemic infants were treated with a novel oral supplement daily: yeast genetically modified to express ferritin. Fecal specimens were analyzed using 16S ribosomal RNA (rRNA) gene amplicon sequencing. Bacterial species richness in anemic infants was not different from that of iron-sufficient infants, but beta diversity and LEfSe analyses of bacterial composition indicated that the microbiota profiles were associated with iron status. Both systemic and oral iron increased alpha and beta diversity metrics. The relative abundance of Ruminococcaceae and other Firmicutes shifted in the direction of an iron-sufficient host, but many different bacteria, including Mollicutes, Tenericutes, and Archaea, were also enriched. Collectively, the findings affirm the important influence of the host’s iron status on commensal bacteria in the gut and concur with clinical concerns about the possibility of adverse consequences after iron supplementation in low-resource settings where children may be carriers of iron-responsive bacterial pathogens. Full article
(This article belongs to the Special Issue Gut Microbiome in Homeostasis and Disease, 3rd Edition)
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