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19 pages, 3244 KB  
Article
Lactobacillus and Bacillus Improve Egg Production in Zhedong White Geese via Gut Microbiota–Metabolite–Endocrine Axis Modulation
by Ruilong Song, Biao Wang, Wan Zhang, Xiao Zhou, Shuyan Rui, Qi Wang, Hehuan Li, Xishuai Tong, Hui Zou, Yonggang Ma, Shufang Chen and Zongping Liu
Vet. Sci. 2026, 13(5), 479; https://doi.org/10.3390/vetsci13050479 (registering DOI) - 15 May 2026
Abstract
Enhancing egg production in geese without antibiotics remains a challenge in poultry science. This study compared the effects of Lactobacillus (LAB) and Bacillus (BAC) probiotics on laying performance, gut microbiota, and serum metabolism in Zhedong White geese. Birds were fed a control diet [...] Read more.
Enhancing egg production in geese without antibiotics remains a challenge in poultry science. This study compared the effects of Lactobacillus (LAB) and Bacillus (BAC) probiotics on laying performance, gut microbiota, and serum metabolism in Zhedong White geese. Birds were fed a control diet or diets supplemented with LAB or BAC. Egg production and quality were monitored throughout the trial. Serum metabolomics and fecal 16S rRNA sequencing were integrated with KEGG enrichment and correlation analyses to uncover functional mechanisms. Both probiotics improved laying performance and egg quality. Total egg production of the LAB group was 8.5% higher than that of the BAC group (p < 0.05). The LAB group’s advantage in egg production was consistent with its stronger activation of the steroid hormone biosynthesis pathway (elevated serum corticosterone and tetrahydrocorticosterone indicated an overall enhancement of steroidogenic flux). Simultaneously, the LAB group exhibited a more efficient conversion of L-phenylalanine to catecholamine precursors, which drove activation of the neuroendocrine reproductive axis. The BAC group showed more significant changes in nitrogen and energy metabolism pathways and a more pronounced expansion of energy-harvesting Firmicutes. These findings reveal two strain-specific regulatory pathways: LAB functions through the “aromatic amino acid–neuroendocrine–steroid hormone axis,” while BAC relies on the “gut microbiota–energy metabolism” pathway, with direct implications for the precise application of probiotics under antibiotic-free farming conditions. Full article
25 pages, 2389 KB  
Article
Probiotic Fermentation Enhances Anti-Diabetic Effects of Chlorella pyrenoidosa in Mice by Modulating Gut Microbiota and Short-Chain Fatty Acids
by Juntao Huang, Yue Zhong, Yang Wu, Siqiao Cai, Wenyu Xiong, Tiantian Li, Bin Liu and Zirui Huang
Foods 2026, 15(10), 1739; https://doi.org/10.3390/foods15101739 - 14 May 2026
Abstract
Numerous studies have demonstrated that Chlorella pyrenoidosa exerts potent anti-diabetic effects, yet its underlying mechanism remains elusive. Notably, no research has explored the anti-diabetic efficacy of its fermented product to date. In this study, we developed a novel compound probiotic-fermented C. pyrenoidosa product [...] Read more.
Numerous studies have demonstrated that Chlorella pyrenoidosa exerts potent anti-diabetic effects, yet its underlying mechanism remains elusive. Notably, no research has explored the anti-diabetic efficacy of its fermented product to date. In this study, we developed a novel compound probiotic-fermented C. pyrenoidosa product (CH-F), and systematically evaluated its anti-diabetic efficacy and mechanism. Furthermore, untargeted metabolomics analysis revealed that fermentation significantly altered the metabolite profile of C. pyrenoidosa, leading to the enrichment of potential anti-diabetic compounds such as acetylcholine and citicoline. In streptozotocin-induced T2DM mice, CH-F effectively ameliorated hyperglycemia, dyslipidemia, and hepatic/pancreatic histological injuries, showing markedly superior efficacy compared to unfermented C. pyrenoidosa. Additionally, CH-F modulated the gut microbiota and improved the levels of short-chain fatty acids (SCFAs) in the intestines of T2DM mice. We demonstrate that CH-F exerts anti-diabetic effects via gut microbiota modulation and SCFA regulation, highlighting its great potential as a functional food ingredient and dietary supplement for the adjuvant management of T2DM. Full article
19 pages, 1730 KB  
Article
Dietary Supplementation of a Multi-Strain Probiotic Increases Muscle Mass in Pigs
by Shu-Hua Hsu, Ting-Yu Lee, Chao-Wei Huang, Bishnu Prasad Bhattarai, Yu-I Pan, Yi-Chu Liao, Hsiao-Tung Chang, Hsin-Hsuan Huang, Jin-Seng Lin, Xin Zhao and Jai-Wei Lee
Int. J. Mol. Sci. 2026, 27(10), 4381; https://doi.org/10.3390/ijms27104381 - 14 May 2026
Abstract
Pork production is closely linked to skeletal muscle growth and anabolic processes. This study investigated the effects of dietary supplementation with a multi-strain probiotic (Lactiplantibacillus plantarum, Streptococcus thermophilus, and Bacillus subtilis) on the growth performance, carcass traits, gut microbiota, [...] Read more.
Pork production is closely linked to skeletal muscle growth and anabolic processes. This study investigated the effects of dietary supplementation with a multi-strain probiotic (Lactiplantibacillus plantarum, Streptococcus thermophilus, and Bacillus subtilis) on the growth performance, carcass traits, gut microbiota, and potential signaling pathways in growing pigs. A total of 144 weaning piglets (28 days old) were randomly allocated to two groups and fed diets with or without probiotics (0.1%) for 18 weeks. Pigs fed with probiotics showed significantly improved feed efficiency (p < 0.05) and greater muscle mass in the loin eye, arm shoulder, and blade shoulder regions. Microbiome analysis revealed significant enrichment of short-chain fatty acid (SCFA)-producing taxa, including Acidaminococcus, Allisonella, Dialister, and Megasphaera, alongside an increased cecal butyrate level in pigs fed probiotics. Integrated fecal microbiome and serum metabolomics analysis demonstrated that the metabolite profile was substantially altered by the supplementation of probiotics. Additionally, serum insulin levels, expression of the bile acid receptor tgr5, and upstream genes in the PI3K/Akt/mTOR pathway (igf1r, insr, and pi3k) were significantly upregulated (p < 0.05). Collectively, these results suggest that a multi-strain probiotic supplementation may be a promising strategy for improving muscle deposition and feed efficiency in commercial pig production. Full article
(This article belongs to the Special Issue Molecular Research in Animal Nutrition)
17 pages, 3862 KB  
Article
Yield Change in Winter Wheat and Rapeseed in Water Shortage Under the Influence of Plant Growth-Promoting Microorganisms and Calcium
by Mariam Zareyan, Rima Mockevičiūtė, Virgilija Gavelienė, Jose Luis Araus, Sigita Jurkonienė and Vaidevutis Šveikauskas
Agronomy 2026, 16(10), 969; https://doi.org/10.3390/agronomy16100969 (registering DOI) - 13 May 2026
Viewed by 4
Abstract
Due to drought stress caused by climate change, a growing global population, and limited land resources, interest in sustainable agriculture is growing. In this study, we evaluate the impact of commercial plant-based probiotics, several beneficial microorganisms, and calcium salts on the growth and [...] Read more.
Due to drought stress caused by climate change, a growing global population, and limited land resources, interest in sustainable agriculture is growing. In this study, we evaluate the impact of commercial plant-based probiotics, several beneficial microorganisms, and calcium salts on the growth and yield of winter wheat and winter rapeseed under limited water resources. The study was conducted in field conditions in two countries simultaneously with different climatic conditions: Spain and Lithuania. Soil was supplemented with calcium in two forms: CaCO3 and CaCl2. Seeds were treated with microorganisms before sowing, and plants were sprayed with them in the spring. The plants inoculated with beneficial microorganisms showed improvement in yield, with harvest index increasing by 5–10% in treated plants. Grain yield was enhanced across treatments, with ProbioHumus + CaCO3 showing the highest yield in Lithuania. Additionally, treated plants exhibited significantly lower stress indicators, with Bacillus subtilis + CaCl2 decreasing lipid peroxidation by 27%. This study provides further evidence that plant treatment with beneficial microorganisms and calcium can contribute to a more environmentally sustainable agriculture. Full article
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27 pages, 6767 KB  
Article
Early-Life Iron Exposure Influences Long-Term Gut Microbiota Recovery After Intestinal Dysbiosis
by Thibault Maumy, Claire McCartney, Ayodeji Samuel Ajayi, Claire Gerkins, Gabriela Fragoso, Annie Calvé and Manuela M. Santos
Microorganisms 2026, 14(5), 1105; https://doi.org/10.3390/microorganisms14051105 - 13 May 2026
Viewed by 70
Abstract
Host–microbiota interactions during the neonatal window of opportunity have gained significant interest as factors influencing long-term health. Factors such as nutrient availability may shape the microbial community, and iron is no exception to this rule. Although the use of iron supplementation is widespread [...] Read more.
Host–microbiota interactions during the neonatal window of opportunity have gained significant interest as factors influencing long-term health. Factors such as nutrient availability may shape the microbial community, and iron is no exception to this rule. Although the use of iron supplementation is widespread during infancy, this micronutrient is known to have profound effects on gut microbiota. This study aimed to determine how early-life iron supplementation shapes gut microbiota composition and whether it influences recovery from gut dysbiosis later in life. Three-week-old female C57BL/6 mice were fed an iron-excess diet for five weeks during the critical period of microbiota establishment. After a two-week washout period to normalize luminal iron content, dysbiosis was induced using either dextran sulfate sodium-induced acute colitis or antibiotic treatment. Mice were then allowed an 8-week recovery period. Gut microbiota composition was longitudinally analyzed by 16S rRNA gene sequencing of fecal samples. Early-life iron supplementation induced durable alterations in gut microbiota composition, with differences persisting even after luminal iron normalization (β-diversity, PERMANOVA p < 0.01). At the endpoint, mice exposed to an iron-sufficient diet remained significantly more distant from their baseline compared to the excess iron group in both the dextran sulfate sodium (33% greater distance) and antibiotic (41% greater distance) models (both p < 0.05). Notably, this pattern was not observed when supplementation occurred in adulthood. In the dextran sulfate sodium model, mice that received an iron-sufficient diet during early life showed an expansion of the probiotic Ligilactobacillus murinus, which positively correlated with fecal succinate levels. Conversely, in the antibiotic-induced model, early-life exposure to an iron-sufficient diet was associated with a more pronounced dysbiosis characterized by a nearly two-fold-greater loss in α-diversity compared to 500 ppm mice (∆Shannon: 1.98 ± 0.22 vs. 1.02 ± 0.25, p < 0.01). These findings suggest that early-life iron supplementation influences long-term host–microbiota interactions and recovery from gut dysbiosis. Full article
(This article belongs to the Special Issue Effects of Diet and Nutrition on Gut Microbiota)
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19 pages, 1446 KB  
Article
The Effect of Prebiotic (Inulin) Addition and Storage Time on the Fatty Acid Profile, Mineral Content, and Sensory Properties of Probiotic Fermented Drinks Produced from Cow’s Milk with Different Fat Contents
by Beata Paszczyk, Marika Bielecka and Jarosław Kowalik
Appl. Sci. 2026, 16(10), 4838; https://doi.org/10.3390/app16104838 - 13 May 2026
Viewed by 80
Abstract
This study evaluated the effect of inulin addition and storage time on the chemical composition, fatty acid profile, mineral content, volatile compounds, and sensory properties of fermented milk drinks produced from cow’s milk with different fat contents (2% and 4%) using the probiotic [...] Read more.
This study evaluated the effect of inulin addition and storage time on the chemical composition, fatty acid profile, mineral content, volatile compounds, and sensory properties of fermented milk drinks produced from cow’s milk with different fat contents (2% and 4%) using the probiotic strain Bifidobacterium animalis subsp. lactis BB-12. Four drink variants were produced: control drinks and drinks supplemented with 2% inulin. Analyses were conducted over 21-day refrigerated storage. The results showed that fat standardization led to significant differences in fat content, whereas protein levels remained relatively stable across samples. The addition of inulin significantly increased dry matter content and improved texture-related sensory attributes, including viscosity, creaminess, and smoothness. GC–IMS analysis revealed that fermentation and storage led to a progressive increase in the contents of volatile compounds, including esters, alcohols, and ketones, with the most complex aroma profile observed after 14 days. Samples with the higher fat content and inulin addition exhibited a greater diversity and intensity of volatile compounds compared to the control drinks. In turn, storage time influenced fatty acid composition, including CLA content, and caused fluctuations in mineral concentrations. Additionally, inulin addition and a higher fat content positively affected the survival of Bifidobacterium animalis subsp. lactis BB-12 during storage. The results indicate that the combined application of inulin and an increased fat content enhances the functional and sensory quality of fermented milk drinks, demonstrating the potential of synbiotic formulations in dairy product development. Full article
(This article belongs to the Special Issue Development and Research of Novel Food Products)
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24 pages, 859 KB  
Article
Bifidobacterium animalis subsp. lactis 832 Alleviates DSS-Induced Colitis in a Murine Model by Regulating Gut Microbiota and Phospholipid Metabolism
by Xintong Chen, Qiushi Wang, Xiaoya Guo, Dan Li, Xinyu Wu, Xiaoya Li, Xiaoyu Zheng, Yangyang Li, Shuangshuang Han, Lu Feng, Bin Liu and Lei Wang
Microorganisms 2026, 14(5), 1090; https://doi.org/10.3390/microorganisms14051090 - 11 May 2026
Viewed by 120
Abstract
Inflammatory bowel disease (IBD) is a chronic intestinal disorder with recurrent inflammation for which effective therapeutic options remain limited. Probiotics from the Bifidobacterium genus have potential beneficial effects on the prevention of IBD by improving intestinal barrier integrity and modulating immune responses. However, [...] Read more.
Inflammatory bowel disease (IBD) is a chronic intestinal disorder with recurrent inflammation for which effective therapeutic options remain limited. Probiotics from the Bifidobacterium genus have potential beneficial effects on the prevention of IBD by improving intestinal barrier integrity and modulating immune responses. However, whether these effects are mediated by the regulation of gut metabolism remains largely unclear. This study was designed to explore the protective effect of an infant-derived Bifidobacterium animalis subsp. lactis 832 (B. lactis 832) on dextran sulfate sodium (DSS)-induced colitis in mice and its underlying mechanism. B. lactis 832 treatment significantly alleviated colitis severity (p < 0.05), as evidenced by reduced weight loss, disease activity index (DAI), and colonic injury, accompanied by significantly decreased pro-inflammatory cytokine expression and increased Il10 expression (p < 0.05). It also improved intestinal barrier integrity and modulated gut microbiota composition by reducing potentially pathogenic bacteria while enriching beneficial taxa. Surprisingly, metabolomic analysis revealed that B. lactis 832 intervention enhanced intestinal phospholipid metabolism, particularly increasing phosphatidylethanolamine (PE) and phosphatidylcholine (PC) levels. Notably, PE or PC supplementation recapitulated the protective effects against DSS-induced colitis (p < 0.05). These findings suggest that B. lactis 832 alleviates colitis through microbiota-associated metabolic regulation, highlighting a key role for phospholipid metabolism in mediating probiotic effects. Full article
(This article belongs to the Section Gut Microbiota)
14 pages, 2059 KB  
Article
Prebiotics Enhance Microbiome Recovery Following Antibiotic-Induced Dysbiosis
by Paige Ramkissoon, Anthony DuCasse, Isaac Berman, Jonathan Sadanaga, Ian O’Neill and David C. Gondek
Microorganisms 2026, 14(5), 1079; https://doi.org/10.3390/microorganisms14051079 - 11 May 2026
Viewed by 313
Abstract
Antibiotic-induced dysbiosis disrupts gut microbiome diversity and functionality, often leading to negative health outcomes, including reduced short-chain fatty acid production, increased susceptibility to opportunistic pathogens, and an increased number of bacterial colonies exhibiting antibiotic resistance. This study investigates the effects of prebiotics (inulin-type [...] Read more.
Antibiotic-induced dysbiosis disrupts gut microbiome diversity and functionality, often leading to negative health outcomes, including reduced short-chain fatty acid production, increased susceptibility to opportunistic pathogens, and an increased number of bacterial colonies exhibiting antibiotic resistance. This study investigates the effects of prebiotics (inulin-type fructans) and probiotic supplementation on microbiome recovery in a murine model. Broad spectrum antibiotics induced near-total microbiome depletion, significantly reducing microbial diversity and metabolite production. Prebiotic supplementation demonstrated superior efficacy during recovery in restoring microbiome diversity (~180 species), improving microbiome diversity metrics, and promoting metabolites, particularly butyrate and valerate, compared to probiotics or unmanipulated recovery. While effective in suppressing opportunistic bacterial growth, probiotics significantly delayed total microbial diversity recovery and resulted in lower diversity metrics (~50 species). However, prebiotic-treated microbiomes exhibited a wider antibiotic resistance profile in culturable bacteria, highlighting prebiotics’ unique impact on the resistome. These findings underscore the potential of prebiotics for recovery from gut dysbiosis while emphasizing the need for further research to address safety considerations regarding their impacts on antibiotic resistance. Importance: This study explores the impact of prebiotic vs. probiotic manipulation of the microbiome in an antibiotic-induced dysbiosis mouse model. Our data demonstrate that prebiotics are more efficacious at enhancing total diversity and limiting the expansion of potentially harmful opportunist bacteria. This is the first study to indicate that prebiotics increase the number of culturable bacterial colonies resistant to antibiotics. These results contribute to our understanding of microbiome manipulation to promote health and limit disease. Full article
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26 pages, 2029 KB  
Systematic Review
Effects of Animal-Based Foods on Metabolic Outcomes in Adults with MASLD and Comorbidities: A Systematic Review of Randomized Controlled Trials (2020–2026)
by Joanna Michalina Jurek, Katarzyna Zablocka-Slowinska, Joanna Pieczynska, Anna Lipert, Helena Clavero-Mestres and Teresa Auguet
Nutrients 2026, 18(10), 1508; https://doi.org/10.3390/nu18101508 - 8 May 2026
Viewed by 494
Abstract
Background: Metabolically dysfunction-associated steatotic liver disease (MASLD) is a growing public health challenge linked to obesity and metabolic dysregulation. Since pharmacological options are limited, Mediterranean (MED) and DASH patterns are recommended. These diets include animal-derived foods, providing essential nutrients but also potential saturated [...] Read more.
Background: Metabolically dysfunction-associated steatotic liver disease (MASLD) is a growing public health challenge linked to obesity and metabolic dysregulation. Since pharmacological options are limited, Mediterranean (MED) and DASH patterns are recommended. These diets include animal-derived foods, providing essential nutrients but also potential saturated fats. Objective: This updated systematic review aims to evaluate evidence from randomized controlled trials (RCTs) regarding the effects of specific animal-based foods on metabolic, hepatic, and anthropometric outcomes in adults with MASLD. Methods: A systematic search of nine databases (including PubMed, Scopus, and Embase) and trial registries (ClinicalTrials.gov) was conducted for RCTs published between January 2020 and 31 March 2026. Participants were adults (18–65 years) with MASLD. Interventions included animal-derived foods typical of MED/DASH. Outcomes focused on hepatic function, lipid profiles, glycemic control, and anthropometry. Risk of bias was assessed using the Cochrane RoB 2.0 tool. Results: A total of seven RCTs demonstrated food-specific and heterogeneous effects. Freshwater fish and vitamin D-fortified probiotic yogurt showed consistent benefits for glycemic control and lipid profiles. Fish oil and omega-3 supplementation led to modest improvements in lipids and hepatic markers. Ghee intake was linked with improved total cholesterol and liver enzyme (ALP). Red meat demonstrated heterogeneous effects depending on dose and background dietary pattern. Most studies exhibited limited concerns regarding risk of bias. Discussion: Animal-based foods within MED/DASH patterns exert distinct effects in MASLD. Freshwater fish and fortified yogurt appear most beneficial. The evidence is limited by small sample sizes in specific food categories and variability in dietary assessment methods. These findings support nuanced, evidence-based recommendations for MASLD management. Full article
(This article belongs to the Section Nutrition and Metabolism)
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17 pages, 6587 KB  
Article
Weizmannia coagulans Long45 Supplementation Prevents Feline-Derived Shigella flexneri 13-Induced Colitis in Mice by Regulation the Nrf2 and NF-κB Signaling Pathways
by Xinyu Zhang, Yuhe Ma, Haozhen Liu, Yang Yang, Yao Ge, Yinfeng Chen, Ying Yang, Jun Lu and Zhenlong Wu
Nutrients 2026, 18(10), 1486; https://doi.org/10.3390/nu18101486 - 7 May 2026
Viewed by 228
Abstract
Background: Shigellosis is an illness that affects young children all over the world and Shigella flexneri is one of the most common pathogens. The aim of this study was to investigate a potential beneficial effect of Weizmannia coagulans Long45 supplementation on feline-derived Shigella [...] Read more.
Background: Shigellosis is an illness that affects young children all over the world and Shigella flexneri is one of the most common pathogens. The aim of this study was to investigate a potential beneficial effect of Weizmannia coagulans Long45 supplementation on feline-derived Shigella flexneri-induced colitis in mice, as well as potential mechanisms. Results: The results revealed that mice receiving fecal microbiota from diarrheic cats experienced significant weight loss, decreased survival rate, increased mRNA levels of inflammatory cytokines (TNF-α, IL-4, IFN-γ, IL-1β, IL-6, and IL-18), and increased cell apoptosis compared to the single DSS treatment. In contrast, mice that received fecal microbiota from healthy cats exhibited an increased body weight, increased mRNA level of ZO-1, claudin-3, and Muc2 and decreased apoptosis, indicating a protective effect. The 16S rDNA analysis revealed that the abundance of Shigella in the feces of diarrheic cats was significantly higher than that in healthy cats, while the abundance of Bacillus was lower. Using bacteria culture technology, 19 strains of Shigella flexneri were isolated from 27 fecal samples of diarrheic cats and a strain of Weizmannia coagulans Long45 was isolated from the feces of healthy cats. Further study showed that Weizmannia coagulans Long45 significantly alleviated pathological alterations and colonic barrier dysfunction by modulating the NF-κB and Nrf2 signaling pathways. Conclusions: Our data indicate that feline-derived Shigella flexneri may be a potential pathogen associated with diarrhea and intestinal barrier dysfunction. Weizmannia coagulans Long45, as a potential probiotic, can effectively alleviate Shigella-induced colitis by interfering with the Nrf2 and NF-κB signaling pathways. Full article
(This article belongs to the Section Nutritional Immunology)
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21 pages, 10421 KB  
Article
Butyrate Is Associated with the Antidepressant Effects of Weizmannia coagulans BC99: Functional Similarity of a Microbial Metabolite in the Microbiota–Gut–Brain Axis
by Yiqing Zhou, Yuwan Li, Shanshan Tie, Yao Dong, Shuguang Fang, Ying Wu and Shaobin Gu
Int. J. Mol. Sci. 2026, 27(9), 4082; https://doi.org/10.3390/ijms27094082 - 2 May 2026
Viewed by 249
Abstract
Butyrate, a short-chain fatty acid derived from the gut microbiota, has been linked to depression through correlational studies; however, whether it might act as a sufficient downstream mediator of the antidepressant effects of a probiotic remains poorly understood. To explore this, a chronic [...] Read more.
Butyrate, a short-chain fatty acid derived from the gut microbiota, has been linked to depression through correlational studies; however, whether it might act as a sufficient downstream mediator of the antidepressant effects of a probiotic remains poorly understood. To explore this, a chronic unpredictable mild stress (CUMS) rat model was established to evaluate the potential antidepressant effects of Weizmannia coagulans BC99. Behavioral assessments included the sucrose preference test (SPT), forced swim test (FST), tail suspension test (TST), and open field test (OFT). In addition, 16S rRNA sequencing, serum metabolomics, and short-chain fatty acid (SCFA) profiling were performed. Levels of inflammatory cytokines (IL-1β, IL-6, IL-4, and LPS) and brain-derived neurotrophic factor (BDNF) were measured in serum, hippocampus, and colon by ELISA. An independent sodium butyrate supplementation experiment was conducted to test functional sufficiency, and hippocampal BDNF/TrkB/CREB signaling was assessed by Western blotting. Treatment with BC99 was associated with alleviation of CUMS-induced depressive-like behaviors, increased butyrate levels, reduced neuroinflammation (IL-1β, IL-6, LPS, and IL-4), and restored hippocampal BDNF levels. BC99 also enriched butyrate-producing bacterial taxa (e.g., Lactobacillus, Bifidobacterium, Faecalibaculum) and normalized tryptophan and sphingolipid metabolism. Notably, sodium butyrate alone recapitulated several of the behavioral and anti-inflammatory effects observed with BC99 and, as shown by Western blot, partially restored hippocampal BDNF/TrkB/CREB signaling, which was impaired in CUMS rats. Together, these findings suggest that butyrate may be associated with the antidepressant effects of W. coagulans BC99, potentially acting through suppression of neuroinflammation and activation of the BDNF pathway. Our results support further investigation of butyrate-enhancing strategies as a nutritional approach for depression. Full article
(This article belongs to the Special Issue Mechanistic Insights into Next-Generation Psychobiotics)
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16 pages, 13501 KB  
Article
Probiotic-Induced Gut Microbiota Modulation: A Comparative Analysis Using 16S rRNA V3–V4 and Targeted Sequencing
by Han Lee, Gaeun Kim, Jungeun Kim, OneZoong Kim, Sung-Hee Jung, Sunghee Hyun and Chang Seok Oh
Microorganisms 2026, 14(5), 1035; https://doi.org/10.3390/microorganisms14051035 - 1 May 2026
Viewed by 489
Abstract
Standard 16S rRNA V3–V4 sequencing encounters primer mismatch issues and insufficient taxonomic resolution, hindering the accurate quantification of specific, low-abundance taxa, such as administered probiotic strains. Therefore, we empirically compared outcomes between the standard V3–V4 method and high-resolution targeted species sequencing (TSS) to [...] Read more.
Standard 16S rRNA V3–V4 sequencing encounters primer mismatch issues and insufficient taxonomic resolution, hindering the accurate quantification of specific, low-abundance taxa, such as administered probiotic strains. Therefore, we empirically compared outcomes between the standard V3–V4 method and high-resolution targeted species sequencing (TSS) to assess bias and establish reliability metrics for probiotic efficacy assessments. A longitudinal pilot study was conducted over nine weeks in older participants receiving synbiotic supplementation; their fecal samples were collected and analyzed. V3–V4 analysis successfully captured a significant transient reduction in alpha-diversity with multidirectional genus-level fluctuations. However, taxonomic overlap between these two methods was high at the phylum level and sharply declined to 6.7% at the species level. Notably, compared with V3–V4 sequencing, TSS could successfully quantify the abundance of administered Bifidobacterium animalis. This study empirically demonstrated that 16S rRNA V3–V4 sequencing introduces substantial quantitative bias, limiting its suitability for monitoring specific probiotic strains and compromising the reliability of clinical efficacy assessments. Therefore, we recommend a dual-sequencing framework that integrates the broad ecological screening capabilities of V3–V4 with the precise species-level quantification of TSS to establish the necessary scientific rigor for assessing probiotic efficacy. Full article
(This article belongs to the Collection Feature Papers in Gut Microbiota Research)
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20 pages, 3091 KB  
Article
Effects of 25-Hydroxyvitamin D3 Combined with Phytase and Probiotic on Calcium–Phosphorus Metabolism, Bone Development, and Growth Performance in Weaned Piglets
by Baoshi Shi, Saiming Gong, Jingjing Wang, Yuyue Xi, Zhiru Tang, Jingchun Gao, Yetong Xu and Zhihong Sun
Nutrients 2026, 18(9), 1428; https://doi.org/10.3390/nu18091428 - 30 Apr 2026
Viewed by 257
Abstract
Background/Objectives: Calcium–phosphorus metabolism is critical for skeletal development in weaned piglets. This study evaluated the effects of dietary 25-hydroxyvitamin D3 (25-OH-VD3) in combination with phytase and probiotics on mineral metabolism, bone development, and related molecular mechanisms in weaned piglets. Methods: [...] Read more.
Background/Objectives: Calcium–phosphorus metabolism is critical for skeletal development in weaned piglets. This study evaluated the effects of dietary 25-hydroxyvitamin D3 (25-OH-VD3) in combination with phytase and probiotics on mineral metabolism, bone development, and related molecular mechanisms in weaned piglets. Methods: Sixty 28-day-old weaned piglets (7.1 ± 1.30 kg) were randomly assigned to four dietary treatments for 31 days (including 3 days of acclimation): CON (basal diet + 50 µg/kg 25-OH-VD3), HI (CON + 50 mg/kg phytase), CY (CON +10 mg/kg probiotics), HICY (CON + 50 mg/kg phytase + 10 mg/kg probiotics). Apparent calcium digestibility, serum biochemical indices, bone mineral density (BMD), and mRNA and protein expression of calcium–phosphorus transport- and metabolism-related genes in jejunal mucosa and kidney were assessed. Results: Compared with CON, piglets in the HI, CY, and HICY groups showed higher apparent calcium digestibility (p < 0.05). Serum transforming growth factor-β was elevated in CY and HICY (p < 0.05). HI enhanced metatarsal and toe BMD (p < 0.05) and upregulated jejunal solute carrier family 34, member 2 (SLC34A2) and SLC34A3 mRNA expression (p < 0.05). In contrast, HICY reduced mRNA expression of transient receptor potential cation channel subfamily V member 6 and calcium-binding protein D28k, as well as of calcium-binding protein D9k and cytochrome P450 27B1 in the kidney (p < 0.05). Renal calcium-sensing receptor protein abundance increased in CY (p < 0.05). Conclusions: Supplementation of 25-OH-VD3 with phytase and/or probiotics improved calcium utilization and modulated key transport pathways, contributing to enhanced bone development in weaned piglets. These findings highlight coordinated nutritional regulation of mineral metabolism during early post-weaning growth. Full article
(This article belongs to the Special Issue Bone-Health-Promoting Bioactive Nutrition)
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21 pages, 2478 KB  
Article
Dietary Probiotics Modulate Oxidative Stress, Metabolic Status, and Immune-Related Gene Expression in Nile Tilapia (Oreochromis niloticus) Exposed to Malathion
by Abdullah A. A. Alghamdi
Vet. Sci. 2026, 13(5), 441; https://doi.org/10.3390/vetsci13050441 - 30 Apr 2026
Viewed by 348
Abstract
Malathion, a widely used organophosphate pesticide, frequently contaminates aquatic ecosystems and poses considerable toxic risks to non-target organisms, including fish. The present study provides an integrated evaluation of the protective effects of dietary probiotics against malathion-associated oxidative, metabolic, and immune-related disturbances in Nile [...] Read more.
Malathion, a widely used organophosphate pesticide, frequently contaminates aquatic ecosystems and poses considerable toxic risks to non-target organisms, including fish. The present study provides an integrated evaluation of the protective effects of dietary probiotics against malathion-associated oxidative, metabolic, and immune-related disturbances in Nile tilapia at the biochemical and molecular levels. After determining the 96 h LC50 of malathion, fish were exposed to a sublethal concentration for 7 days followed by a 14-day recovery period while receiving either a basal or probiotic-supplemented diet. Malathion exposure increased cumulative mortality, induced behavioral stress, and caused metabolic and hepatorenal disturbances characterized by elevated serum glucose and cholesterol, altered serum protein fractions, increased alanine and aspartate aminotransferase activities, and elevated creatinine and uric acid levels. Oxidative stress was evidenced by increased serum malondialdehyde and transcriptional suppression of antioxidant-related genes (sod-2 and cat) in the liver, spleen, and intestine. Malathion also triggered immune dysregulation through the upregulation of pro-inflammatory cytokine genes (il-1β and tnf-α) and suppression of regulatory cytokines (tgf-β and il-10). Probiotic supplementation during recovery significantly reduced mortality, restored metabolic and hepatorenal biomarkers, attenuated oxidative damage, and enhanced antioxidant capacity at both the biochemical and transcriptional levels. Moreover, probiotic-supplemented fish exhibited controlled pro-inflammatory signaling accompanied by the pronounced activation of regulatory cytokines, indicating balanced immune modulation. Collectively, dietary probiotics effectively mitigate malathion-induced toxicity by improving antioxidant defense, immune regulation, and physiological resilience, highlighting their potential as functional dietary additives for sustainable aquaculture in Nile tilapia. Full article
(This article belongs to the Special Issue Advances in Zoo, Aquatic, and Wild Animal Medicine)
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Article
The Effects of Dietary Enterococcus faecalis HHP003 Supplementation on Gut Microbiota Composition and Gut Health in Cats with Mild Diarrhea
by Qin Wang, Yanyi Zheng, Wenyu Huang, Feitong Liu, Lingling Zhao, Siyuan Xue, Huiwen Zeng and Yi Wu
Animals 2026, 16(9), 1366; https://doi.org/10.3390/ani16091366 - 29 Apr 2026
Viewed by 356
Abstract
As a probiotic lactic acid bacterium, E. faecalis regulates intestinal flora, strengthens the intestinal barrier, and enhances immunity in the host. However, as a new strain isolated from healthy feline gut, E. faecalis HHP003 has an unclear effect on feline intestinal health. Twenty [...] Read more.
As a probiotic lactic acid bacterium, E. faecalis regulates intestinal flora, strengthens the intestinal barrier, and enhances immunity in the host. However, as a new strain isolated from healthy feline gut, E. faecalis HHP003 has an unclear effect on feline intestinal health. Twenty cats with mild diarrhea were randomly assigned to two groups: one group received the standard diet (MD), while the other group received supplemental E. faecalis HHP003 (EF). Meanwhile, 10 healthy cats were enrolled as a healthy control group (CON). After 42 days of intervention, the EF group exhibited significantly reduced serum inflammatory markers (TNF-α and IL-1β), as well as decreased levels of calprotectin and lipopolysaccharide, compared with the MD group (p < 0.05). The intestinal microbial diversity was altered in the EF group, with increased Chao and Shannon indices (p < 0.05). Specifically, the relative abundances of Bacillota, Bacteroidota, and Ruminococcaceae were significantly higher in the EF group than those in the MD group (p < 0.05). Metabolomic analysis identified 697 differential metabolites and nine KEGG metabolic pathways (p < 0.05) between the EF and MD groups. Furthermore, the study identified significant associations between the gut microbiota and selected serum metabolites. In summary, E. faecalis HHP003 supplementation was associated with reduced serum inflammatory responses, improved intestinal inflammation and barrier markers, and altered gut microbiota and serum metabolite levels in cats with mild diarrhea. Full article
(This article belongs to the Special Issue Dietary Supplement in Companion Animals)
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