Intestinal Mucosal Microbiota Research Articles

Zhishi Daozhi decoction alleviates constipation induced by a high-fat and high-protein diet via regulating intestinal mucosal microbiota and oxidative stress.

A growing body of evidence has demonstrated that a high-fat and high-protein diet (HFHPD) causes constipation. This study focuses on understanding how the use of Zhishi Daozhi decoction (ZDD) affects the intricate balance of intestinal microorganisms. The insights gained from this investigation hold the potential to offer practical clinical approaches to mitigate the constipation-related issues associated with HFHPD. Mice were randomly divided into five groups: the normal (MN) group, the natural recovery (MR) group, the low-dose ZDD (MLD) group, the medium-dose ZDD (MMD) group, and the high-dose ZDD (MHD) group. After the constipation model was established by HFHPD combined with loperamide hydrochloride (LOP), different doses of ZDD were used for intervention. Subsequently, the contents of cholecystokinin (CCK) and calcitonin gene-related peptide (CGRP) in serum, superoxide dismutase (SOD), and malondialdehyde (MDA) in the liver were determined. The DNA of intestinal mucosa was extracted, and 16S rRNA amplicon sequencing was used to analyze the changes in intestinal mucosal microbiota. After ZDD treatment, CCK content in MR group decreased and CGRP content increased, but the changes were not significant. In addition, the SOD content in MR group was significantly lower than in MLD, MMD, and MHD groups, and the MDA content in MR group was significantly higher than in MN, MLD, and MHD groups. Constipation modeling and the intervention of ZDD changed the structure of the intestinal mucosal microbiota. In the constipation induced by HFHPD, the relative abundance of pathogenic bacteria such as Aerococcus, Staphylococcus, Corynebacterium, Desulfovibrio, Clostridium, and Prevotella increased. After the intervention of ZDD, the relative abundance of these pathogenic bacteria decreased, and the relative abundance of Candidatus Arthromitus and the abundance of Tropane, piperidine, and pyridine alkaloid biosynthesis pathways increased in MHD group. Constipation induced by HFHPD can increase pathogenic bacteria in the intestinal mucosa, while ZDD can effectively relieve constipation, reduce the relative abundance of pathogenic bacteria, and alleviate oxidative stress injury. In addition, high-dose ZDD can increase the abundance of beneficial bacteria, which is more conducive to the treatment of constipation.

Feed Regime Slightly Modifies the Bacterial but Not the Fungal Communities in the Intestinal Mucosal Microbiota of Cobia Fish (Rachycentron canadum).

The bacterial community of the intestinal microbiota influences many host functions, and similar effects have been recently reported for the fungal community (mycobiota). Cobia is a tropical fish that has been studied for its potential in marine aquaculture. However, the study of its bacterial community has been underreported and the mycobiota has not been investigated. We analyzed the gut bacterial and fungal profile present in the intestinal mucosa of reared adult cobias fed two diets (frozen fish pieces (FFPs) and formulated feed (FF)) for 4 months by sequencing the 16S rRNA (V3-V4) and internal transcribed spacer-2 (ITS2) regions using Illumina NovaSeq 6000. No significant differences in the alpha diversity of the bacterial community were observed, which was dominated by the phyla Proteobacteria (~96%) and Firmicutes (~1%). Cobia fed FF showed higher abundance of 10 genera, mainly UCG-002 (Family Oscillospiraceae) and Faecalibacterium, compared to cobia fed FFPs, which showed higher abundance of 7 genera, mainly Methylobacterium-Methylorubrum and Cutibacterium. The inferred bacterial functions were related to metabolism, environmental information processing and cellular processes; and no differences were found between diets. In mycobiota, no differences were observed in the diversity and composition of cobia fed the two diets. The mycobiota was dominated by the phyla Ascomycota (~88%) and Basidiomycota (~11%). This is the first study to describe the gut bacterial and fungal communities in cobia reared under captive conditions and fed on different diets and to identify the genus Ascobulus as a new member of the core fish mycobiota.

Diarrhea accompanies intestinal inflammation and intestinal mucosal microbiota dysbiosis during fatigue combined with a high-fat diet

ObjectiveIt was reported fatigue or a high-fat diet triggers diarrhea, and intestinal microbiota may play central roles in diarrhea. Therefore, we investigated the association between the intestinal mucosal microbiota and the intestinal mucosal barrier from fatigue combined with a high-fat diet.MethodThis study divided the Specific pathogen-free (SPF) male mice into the normal group (MCN) and the standing united lard group (MSLD). The MSLD group stood on water environment platform box for 4 h/day for 14 days, and 0.4 mL lard was gavaged from day 8, twice daily for 7 days.ResultAfter 14 days, Mice in the MSLD group showed diarrhea symptoms. The pathological analysis showed structural damage to the small intestine in the MSLD group, with an increasing trend of interleukin-6 (IL-6) and IL-17, and inflammation accompanied by structural damage to the intestine. Fatigue combined with a high-fat diet considerably decreased Limosilactobacillus vaginalis and Limosilactobacillus reuteri, and among them, Limosilactobacillus reuteri positively associated with Muc2 and negatively with IL-6.ConclusionThe interactions between Limosilactobacillus reuteri and intestinal inflammation might be involved in the process of intestinal mucosal barrier impairment in fatigue combined with high-fat diet-induced diarrhea.

Brain-bacteria-gut axis and oxidative stress mediated by intestinal mucosal microbiota might be an important mechanism for constipation in mice.

Intestinal microbiota disorder was associated with constipation. This study investigated the microbiota-gut-brain axis and oxidative stress mediated by intestinal mucosal microbiota in mice with spleen deficiency constipation. The Kunming mice were randomly divided into the control (MC) group and the constipation (MM) group. The spleen deficiency constipation model was established by gavage with Folium sennae decoction and controlled diet and water intake. The body weight, spleen and thymus index, 5-Hydroxytryptamine (5-HT) and Superoxide Dismutase (SOD) content were significantly lower in the MM group than the MC group, the content of vasoactive intestinal peptide (VIP) and malondialdehyde (MDA) content were significantly higher than the MC group. The Alpha diversity of intestinal mucosal bacteria was not changed but beta diversity was changed in mice with spleen deficiency constipation. Compared to the MC group, the relative abundance of Proteobacteria was an upward trend and the Firmicutes/Bacteroidota (F/B) value was a downward trend in the MM group. There was a significant difference in the characteristic microbiota between the two groups. In the MM group, Brevinema, Akkermansia, Parasutterella, Faecalibaculum, Aeromonas, Sphingobium, Actinobacillus, and other pathogenic bacteria were enriched. Meanwhile, there was a certain relationship between the microbiota and gastrointestinal neuropeptide and oxidative stress indicators. The community structure of intestinal mucosal bacteria in mice with spleen deficiency constipation was changed, which was characterized by the reduction of F/B value and enrichment of Proteobacteria. Microbiota-gut-brain axis may be important for spleen deficiency constipation.

Long-term consumption of different doses of Grifola frondosa affects immunity and metabolism: correlation with intestinal mucosal microbiota and blood lipids.

Grifola frondosa (GF) is an edible mushroom with hypoglycemic and hypolipidemic effects. In this study, the specific pathogen-free male mice were randomized into the normal (NM), low-dose GF (LGF), medium-dose GF (MGF), and high-dose GF (HGF) groups. The LGF, MGF, and HGF groups were fed with 1.425g/(kgd), 2.85g/(kgd), and 5.735g/(kgd) of GF solution for 8weeks. After feeding with GF solution, compared with the NM group, the thymus index was significantly increased in the LGF group, and TC, TG, and LDL of mice were significantly increased in the HGF group, while HDL was significantly decreased. Compared with the NM group, the uncultured Bacteroidales bacterium, Ligilactobacillus increased in the LGF group, and Candidatus Arthromitus increased in the MGF group. The characteristic bacteria of the HGF group included Christensenellaceae R7, unclassified Clostridia UCG 014, unclassified Eubacteria coprostanoligenes, and Prevotellaceae Ga6A1. Among them, Ligilactobacillus showed a negative correlation with HDL. Unclassified Eubacterium coprostanoligenes group and Ligilactobacillus showed a positive correlation with TG. In summary, our experiments evidenced that GF improves lipid metabolism disorders by regulating the intestinal microbiota, providing a new pathway for hypolipidemic using GF dietary.

The process of hypertension induced by high-salt diet: Association with interactions between intestinal mucosal microbiota, and chronic low-grade inflammation, end-organ damage.

Inflammation and immunity play a major role in the development of hypertension, and a potential correlation between host mucosal immunity and inflammatory response regulation. We explored the changes of intestinal mucosal microbiota in hypertensive rats induced by high-salt diet and the potential link between the intestinal mucosal microbiota and inflammation in rats. Therefore, we used PacBio (Pacific Bioscience) SMRT sequencing technology to determine the structure of intestinal mucosal microbiota, used enzyme-linked immunosorbent assay (ELISA) to determined the proinflammatory cytokines and hormones associated with hypertension in serum, and used histopathology methods to observe the kidney and vascular structure. We performed a potential association analysis between intestinal mucosal characteristic bacteria and significantly different blood cytokines in hypertensive rats induced by high-salt. The results showed that the kidney and vascular structures of hypertensive rats induced by high salt were damaged, the serum concentration of necrosis factor-α (TNF-α), angiotensin II (AngII), interleukin-6 (IL-6), and interleukin-8 (IL-8) were significantly increased (p < 0.05), and the coefficient of immune organ spleen was significantly changed (p < 0.05), but there was no significant change in serum lipids (p > 0.05). From the perspective of gut microbiota, high-salt diet leads to significant changes in intestinal mucosal microbiota. Bifidobacterium animalis subsp. and Brachybacterium paraconglomeratum were the dominant differential bacteria in intestinal mucosal, with the AUC (area under curve) value of Bifidobacterium animalis subsp. and Brachybacterium paraconglomeratum were 1 and 0.875 according to ROC (receiver operating characteristic) analysis. Correlation analysis showed that Bifidobacterium animalis subsp. was correlated with IL-6, IL-8, TNF-α, and Ang II. Based on our results, we can speculated that high salt diet mediated chronic low-grade inflammation through inhibited the growth of Bifidobacterium animalis subsp. in intestinal mucosa and caused end-organ damage, which leads to hypertension.

The alteration of intestinal mucosal α-synuclein expression and mucosal microbiota in Parkinson’s disease

Parkinson’s disease (PD) is the second most common neurodegenerative disease but still lacks a preclinical strategy to identify it. The diagnostic value of intestinal mucosal α-synuclein (αSyn) in PD has not drawn a uniform conclusion. The relationship between the alteration of intestinal mucosal αSyn expression and mucosal microbiota is unclear. Nineteen PD patients and twenty-two healthy controls were enrolled in our study from whom were collected, using gastrointestinal endoscopes, duodenal and sigmoid mucosal samples for biopsy. Multiplex immunohistochemistry was performed to detect total, phosphorylate, and oligomer α-synuclein. Next-generation 16S rRNA amplicon sequencing was applied for taxonomic analysis. The results implied that oligomer α-synuclein (OSyn) in sigmoid mucosa of PD patients was transferred from the intestinal epithelial cell membrane to the cytoplasm, acinar lumen, and stroma. Its distribution feature was significantly different between the two groups, especially the ratio of OSyn/αSyn. The microbiota composition in mucosa also differed. The relative abundances of Kiloniellales, Flavobacteriaceae, and CAG56 were lower, while those of Proteobacteria, Gammaproteobacteria, Burkholderiales, Burkholdriaceae, Oxalobacteraceae, Ralstonia, Massilla, and Lactoccus were higher in duodenal mucosa of PD patients. The relative abundances of Thermoactinomycetales and Thermoactinomycetaceae were lower, while those of Prevotellaceae and Bifidobacterium longum were higher in patients’ sigmoid mucosa. Further, the OSyn/αSyn level was positively correlated with the relative abundances of Proteobacteria, Gammaproteobacteria, Burkholderiales, Pseudomonadales, Burkholderiaceae, and Ralstonia in the duodenal mucosa, while it was negatively correlated with the Chao1 index and observed operational taxonomic units of microbiota in sigmoid mucosa. The intestinal mucosal microbiota composition of PD patients altered with the relative abundances of proinflammatory bacteria in the duodenal mucosa increased. The ratio of the OSyn/αSyn level in the sigmoid mucosa indicated a potential diagnostic value for PD, which also correlated with mucosal microbiota diversity and composition.Key points• The distribution of OSyn in sigmoid mucosa differed between PD patients and healthy controls.• Significant alterations in the microbiome were found in PD patients’ gut mucosa.• OSyn/αSyn level in sigmoid mucosa indicated a potential diagnostic value for PD.

Simo decoction curing spleen deficiency constipation was associated with brain-bacteria-gut axis by intestinal mucosal microbiota.

Simo decoction (SMD) is a traditional prescription for treating gastrointestinal diseases. More and more evidences prove that SMD can treat constipation by regulating intestinal microbiota and related oxidative stress indicators, but the specific mechanism is still unclear. A network pharmacological analysis was used to predict the medicinal substances and potential targets of SMD to alleviate constipation. Then, 15 male mice were randomly divided into normal group (MN group), natural recovery group (MR group), and SMD treatment group (MT group). Constipation model mice were constructed by gavage of Folium sennae decoction and control of diet and drinking water, and SMD was used for intervention after successful modeling. The levels of 5-hydroxytryptamine (5-HT), vasoactive intestinal peptide (VIP), superoxide dismutase (SOD), malondialdehyde (MDA), and fecal microbial activities were measured, and the intestinal mucosal microbiota was sequenced. Network pharmacology analysis showed that a total of 24 potential active components were obtained from SMD, and 226 target proteins were obtained after conversion. Meanwhile, we obtained 1,273 and 424 disease-related targets in the GeneCards database and the DisGeNET database, respectively. After combination and deduplication, the disease targets shared 101 targets with the potential active components of SMD. When the mice were intervened with SMD, the 5-HT, VIP, MDA, SOD content, and microbial activity in MT group were close to MN group, and Chao 1 and ACE in MT group were significantly higher than that in MR group. In the Linear discriminant analysis Effect Size (LEfSe) analysis, the abundance of beneficial bacteria such as Bacteroides, Faecalibacterium, Alistipes, Subdoligranulum, Lactiplantibacillus, and Phascolarctobacterium in MT group increased. At the same time, there were some associations between microbiota and brain-gut peptides and oxidative stress indicators. SMD can promote intestinal health and relieve constipation through brain-bacteria-gut axis associating with intestinal mucosal microbiota and alleviate oxidative stress.

The diarrheal mechanism of mice with a high-fat diet in a fatigued state is associated with intestinal mucosa microbiota.

Growing evidence has demonstrated that fatigue and a high-fat diet trigger diarrhea, and intestinal microbiota disorder interact with diarrhea. However, the association of intestinal mucosal microbiota with fatigue and high-fat diet trigger diarrhea remains unclear. The specific pathogen-free Kunming male mice were randomly divided into the normal group (MCN), standing group (MSD), lard group (MLD), and standing united lard group (MSLD). Mice in the MSD and MSLD groups stood on the multiple-platform apparatus for four h/d for fourteen consecutive days. From the eighth day, mice in the MLD and MSLD groups were intragastric lard, 0.4mL/each, twice a day for seven days. Subsequently, we analyzed the characteristics and interaction relationship of intestinal mucosal microbiota, interleukin-6 (IL-6), interleukin-17 (IL-17), malondialdehyde (MDA), superoxide dismutase (SOD), and secretory immunoglobulin A (sIgA). Results showed that mice in the MSLD group had an increased number of bowel movements. Compared with the MCN group, the contents of IL-17, and IL-6 were higher (p > 0.05), and the content of sIgA was lower in the MSLD group (p > 0.05). MDA and SOD increased in MLD and MSLD groups. Thermoactinomyces and Staphyloccus were the characteristic bacteria of the MSLD group. And Staphyloccus were positively correlated with IL-6, IL-17, and SOD. In conclusion, the interactions between Thermoactinomyces, Staphyloccus and intestinal inflammation, and immunity might be involved in fatigue and high-fat diet-induced diarrhea.

Dose of sucrose affects the efficacy of Qiweibaizhu powder on antibiotic-associated diarrhea: Association with intestinal mucosal microbiota, short-chain fatty acids, IL-17, and MUC2.

Due to the poor taste of Qiweibaizhu powder (QWBZP), patients have difficulty taking medicine, which leads to poor compliance and limits clinical use to a certain extent. In the trend of restricting sugar intake, sweeteners have gained massive popularity, among which sucrose is a commonly used sweetener in preparations. This study aimed to investigate the effect of different sucrose dose addition with antibiotic-associated diarrhea (AAD) by intervened QWBZP on intestinal mucosal microbiota. Thirty specific-pathogen-free (SPF) Kunming (KM) male mice were randomly divided into normal group (N), natural recovery group (M), QWBZP group (Q), low dose sucrose group (LQ), medium dose sucrose group (MQ), and high dose sucrose group (HQ). Subsequently, 16S rRNA amplicon sequencing and GC-MS techniques were used to analyze the intestinal mucosal microbiota and short-chain fatty acid (SCFAs) in intestinal contents, respectively, and enzyme-linked immunosorbent assay was used to determine mucin 2 (MUC2) and interleukin 17 (IL-17). Compared with the Q group, the results showed that with the increase of sucrose dose, the intestinal microbial structure of mice was significantly altered, and the intestinal microbial diversity was elevated, with the poor restoration of the intestinal biological barrier, decreased content of SCFAs, high expression of inflammatory factor IL-17 and decreased content of mucosal protective factor MUC2. In conclusion, we found that the addition of sucrose had an effect on the efficacy of the AAD intervented by QWBZP, which was less effective than QWBZP, showing a certain dose-response relationship. In this experiment, it was concluded that the addition of sucrose might also further lead to intestinal inflammation and the disruption of the intestinal mucosal barrier, and the production of metabolites SCFAs. The addition of sucrose might also further lead to intestinal inflammation and the disruption of the intestinal mucosal barrier, and the production of metabolites SCFAs. However, these findings still need to be verified in a more extensive study. The effect of adding the sweetener sucrose on the efficacy of Chinese herbal medicine in treating diseases also still needs more research.

Alteration of intestinal mucosal microbiota in mice with Chinese dampness-heat syndrom diarrhea by improper diet combined with high temperature and humidity environments.

Environment, diet, and emotion may trigger diarrhea, but the mechanism is unclear. Dietary habits or environmental factors affect the composition of gut microbiota. This study aimed to investigate the effects of improper diet combined with high humidity and temperature (HTH) environment on the intestinal mucosal microbiota. Kunming mice were randomly assigned to two equal groups of five mice, namely the control (ccm) group and the model (cmm) group. Diarrhea mice with dampness-heat (DSH) were established by improper diet combined with HTH environments. We used 16S rRNA gene amplicon sequencing to analyze the characteristics of intestinal mucosal microbiota and the interaction relationship of function. Our study shows that the intestinal mucosal microbiota of mice changed significantly after an improper diet combined with the HTH environments. The abundance of Fusobacteria and Haemophilus increased dramatically in the cmm group compared to the ccm group (P<0.05). And the abundance of Firmicutes, Lactobacillus, and Lonsdalea was significantly decreased in the cmm group (P<0.05). According to the functional predictive analysis, we found that Lactobacillus showed a significant negative correlation with Protein export, Homologous recombination, Phenylalanine, tyrosine, tryptophan biosynthesis, Citrate cycle, and Lipoic acid metabolism. Diarrhea with DSH constructed under improper diet and HTH environment may be related to Lactobacillus and Haemophilus. And long-term consumption of improper diet and the HTH environment may affect metabolism.

Significant alterations of intestinal symbiotic microbiota induced by intraperitoneal vaccination mediate changes in intestinal metabolism of NEW Genetically Improved Farmed Tilapia (NEW GIFT, Oreochromis niloticus)

BackgroundAfter millions of years of coevolution, symbiotic microbiota has become an integral part of the host and plays an important role in host immunity, metabolism, and health. Vaccination, as an effective means of preventing infectious diseases, has been playing a vital role in the prevention and control of human and animal diseases for decades. However, so far, minimal is known about the effect of vaccination on fish symbiotic microbiota, especially mucosal microbiota, and its correlation with intestinal metabolism remains unclear.MethodsHere we reported the effect of an inactivated bivalent Aeromonas hydrophila/Aeromonas veronii vaccine on the symbiotic microbiota and its correlation with the intestinal metabolism of farmed adult Nile tilapia (Oreochromis niloticus) by 16S rRNA gene high-throughput sequencing and gas chromatography-mass spectrometry metabolomics.ResultsResults showed that vaccination significantly changed the structure, composition, and predictive function of intestinal mucosal microbiota but did not significantly affect the symbiotic microbiota of other sites including gill mucosae, stomach contents, and stomach mucosae. Moreover, vaccination significantly reduced the relative abundance values of potential opportunistic pathogens such as Aeromonas, Escherichia–Shigella, and Acinetobacter in intestinal mucosae. Combined with the enhancement of immune function after vaccination, inactivated bivalent Aeromonas vaccination had a protective effect against the intestinal pathogen infection of tilapia. In addition, the metabolite differential analysis showed that vaccination significantly increased the concentrations of carbohydrate-related metabolites such as lactic acid, succinic acid, and gluconic acid but significantly decreased the concentrations of multiple lipid-related metabolites in tilapia intestines. Vaccination affected the intestinal metabolism of tilapia, which was further verified by the predictive function of intestinal microbiota. Furthermore, the correlation analyses showed that most of the intestinal differential microorganisms were significantly correlated with intestinal differential metabolites after vaccination, confirming that the effect of vaccination on intestinal metabolism was closely related to the intestinal microbiota.ConclusionsIn conclusion, this paper revealed the microbial and metabolic responses induced by inactivated vaccination, suggesting that intestinal microbiota might mediate the effect of vaccination on the intestinal metabolism of tilapia. It expanded the novel understanding of vaccine protective mechanisms from microbial and metabolic perspectives, providing important implications for the potential influence of vaccination on human intestinal microbiota and metabolism.-tufGxbkxhm-UiFTxxYHNiVideo

Effects of sweeteners on host physiology by intestinal mucosal microbiota: Example-addition sweeteners in Qiweibaizhu Powder on intestinal mucosal microbiota of mice with antibiotic-associated diarrhea.

In recent years, sweeteners have gained massive popularity under the trend of limiting sugar intake. Our previous study found that Qiweibaizhu Powder (QWBZP) could improve gut microbiota dysbiosis and has good efficacy in treating antibiotic-associated diarrhea (AAD). In this study, we investigated the effects of sucrose, sorbitol, xylitol, and saccharin on the intestinal mucosal microbiota of AAD mice treated with QWBZP. When the AAD model was constructed by being gavaged mixed antibiotic solution, Kunming mice were randomly assigned to seven groups: the control (mn) group, the ADD (mm) group, the QWBZP (mq) group, the saccharin + QWBZP (mc) group, the sucrose + QWBZP (ms) group, the xylito + QWBZP (mx) group, and the sorbitol + QWBZP (msl) group. Subsequently, 16S rRNA gene amplicon sequencing was used to analyze the intestinal mucosal microbiota composition and abundance. The results showed that feces from AAD mice were diluted and wet and improved diarrhea symptoms with QWBZP and sorbitol. In contrast, the addition of sucrose, saccharin, and xylitol delayed the healing of diarrhea. The relative abundance of intestinal mucosal microbiota showed Glutamicibacter, Robinsoniella, and Blautia were characteristic bacteria of the mx group, Candidatus Arthromitus, and Bacteroidales_S24-7_group as the typical bacteria of the mn group, Clostridium_innocuum_group as the distinct bacteria of the mm group. Mycoplasma and Bifidobacterium as the characteristic bacteria of the ms group. Correlation analysis of typical bacterial genera with metabolic functions shows that Blautia negatively correlates with D-Glutamine and D-glutamate metabolism. Bacteroidales_S24-7_group has a significant negative correlation with the Synthesis and degradation of ketone bodies. The study confirmed that sucrose, sorbitol, xylitol, and saccharin might further influence metabolic function by altering the intestinal mucosal microbiota. Compared to the other sweetener, adding sorbitol to QWBZP was the best therapeutic effect for AAD and increased the biosynthesis and degradation activities. It provides the experimental basis for applying artificial sweeteners in traditional Chinese medicine (TCM) as a reference for further rational development and safe use of artificial sweeteners.

Distinctions Between Fecal and Intestinal Mucosal Microbiota in Subgroups of Irritable Bowel Syndrome.

Recent studies have shown that changes in the intestinal microbiota contribute to the pathogenesis of irritable bowel syndrome (IBS). This study aimed to investigate the characteristics of the fecal and intestinal mucosal microbiota in IBS patients, and the correlation between microbiota and clinical manifestations. Fecal and intestinal mucosal samples were collected from 14 constipation-predominant IBS (IBS-C) patients, 20 diarrhea-predominant IBS (IBS-D) patients, and 20 healthy controls (HCs). 16S rRNA gene sequencing and fluorescence in situ hybridization were used for the analysis of samples. Community richness and diversity of the fecal microbiota in IBS patients were significantly reduced compared with the HCs. The mucosal samples in IBS patients showed decreased Bifidobacterium and increased Bacteroides caccae compared with HCs; Eubacterium and Roseburia were decreased in IBS-C patients and increased in IBS-D patients. A comparison of the fecal and mucosal microbiota in IBS patients showed significantly increased Bifidobacterium in fecal samples and a decrease in mucosal samples in IBS-C patients; Bacteroides caccae and Roseburia were significantly reduced in fecal samples and increased in mucosal samples of IBS patients. A correlation between microbiota and clinical manifestations in IBS patients showed that Bacteroides caccae and Roseburia in fecal samples and Bifidobacterium and Eubacterium in mucosal samples were associated with abdominal pain and distention. Distinct differences exist between the fecal and intestinal mucosal microbiota in IBS patients, with the changes in the latter appearing more consistent with the pathophysiology of IBS. Changes in intestinal microbiota were associated with the clinical manifestations in IBS.

Diarrheal Mice Induced by a High-fat and High-protein Diet could be Cured by Bohe Pill Regulating Intestinal Mucosal Bacteria

This study aimed to investigate the effect of Baohe pill decoction on the bacterial diversity and community structure of the intestinal mucosa of mice with diarrhea caused by feeding HFD and HPD, and provides a scientific basis for the efficacy of Baohe pill decoction in treating this condition. Specific pathogen-free Kunming male mice were randomized into three equal groups, i.e. the control (ftcm), natural recovery (ftmm), and Baohe pill decoction treatment (fttm) groups. The ftcm group was fed a general feed diet, and the ftmm and fttm groups were treated with HFD and HPD. Following the successful induction of diarrhea, the mice in the fttm group were gavaged with Baohe pill decoction, and the mice in the ftmm group were given equal amounts of sterile water. The intestinal mucosal microbiota characteristics were analyzed by 16S ribosomal ribonucleic acid Pacbio single-molecule, real-time full-length gene sequencing. The operational taxonomic unit (OTU) number and alpha diversity in the intestinal mucosal flora of the fttm group were higher than in the ftcm and ftmm groups (P > 0.05), and the composition of the intestinal mucosa bacteria differed between the ftmm and fttm groups. The ftmm group showed a higher abundance of Proteobacteria, Actinobacteria, and Streptophyta, while the Firmicutes, Bacteroidetes, and Verrucomicrobia in the fttm group were higher than that in the other two groups. Compared with the ftmm group, certain beneficial bacteria (Lactobacillus) showed an increased abundance, and pathogenic bacteria (Bacillus, Bradyrhizobium, and Ralstonia) showed a decreased abundance. Bacterial species, such as Lactobacillus (Lactobacillus johnsonii, Lactobacillus murinus, Lactobacillus acidophilus and Lactobacillus intestinalis) were significantly higher in the fttm and closely matched those in the ftcm group. Furthermore, various opportunistic pathogens caused by feeding an HFD and HPD were not detected in the fttm group. The current study results indicated that Baohe pill decoction improved diarrhea symptoms by regulating the balance of mucosal microbiota. Specifically, Baohe pill decoction increased the abundance of probiotic Lactobacillus and decreased the abundance of the opportunistic Ralstonia pathogen.

The Compositional Structure of the Small Intestinal Microbial Community via Balloon-Assisted Enteroscopy

Introduction: An association has been found between human-gut microbiota and various diseases (e.g., metabolic disease) by analyzing fecal or colonic microbiota. Despite the importance of the small intestinal microbiota, sampling difficulties prevent its full analysis. We investigated the composition and metagenomic functions of microbiota along the small intestine and compared them with the microbiota from feces and from other gastrointestinal (GI) sites. Methods: Mucosal samples from the six GI sites (stomach, duodenum, distal jejunum, proximal ileum, terminal ileum, and rectum) were collected under balloon-assisted enteroscopy. Fecal samples were collected from all participants. The microbial structures and metagenomic functions of the small intestinal mucosal microbiota were compared with those from feces and other GI sites using 16S ribosomal RNA gene sequencing. Results: We analyzed 133 samples from 29 participants. Microbial beta diversity analysis showed that the jejunum and ileum differed significantly from the lower GI tract and the feces (p < 0.001). Jejunal and duodenal microbiotas formed similar clusters. Wide clusters spanning the upper and lower GI tracts were observed with the ileal microbiota, which differed significantly from the jejunal microbiota (p < 0.001). Veillonella and Streptococcus were abundant in the jejunum but less so in the lower GI tract and feces. The metagenomic functions associated with nutrient metabolism differed significantly between the small intestine and the feces. Conclusions: The fact that the compositional structures of small intestinal microbiota differed from those of fecal and other GI microbiotas reveals that analyzing the small intestinal microbiota is necessary for association studies on metabolic diseases and gut microbiota.

Intestinal mucosal microbiota composition of patients with acquired immune deficiency syndrome in Guangzhou, China.

Acquired immune deficiency syndrome, caused by the human immunodeficiency virus (HIV), has been associated with intestinal dysbiosis, which includes an increase in the number of mucosa-associated pathobionts. In the present study, the intestinal mucosal microbiota patterns of HIV-infected patients were compared with those of healthy individuals in a population from Guangzhou, China. The gut microbiota of intestinal mucosal samples from 12 patients with HIV (transmission routes included sex and intravenous drug abuse) was compared with that of 12 healthy age- and sex-matched controls. Gut microbial communities were profiled via sequencing of the bacterial 16S ribosomal RNA genes. Dysbiosis in HIV-infected individuals was characterized by decreased α-diversity, decreased levels of Firmicutes and increased levels of Proteobacteria. Furthermore, low mean counts of Lachnoclostridium, Roseburia, Thauera, Dorea and Roseburia inulinivorans, and high mean counts of Halomonas and Shewanella bacteria, were indicated to be HIV-associated mucosal bacterial alterations. The relative abundance of Fusobacterium and Lachnoclostridium was significantly decreased, while that of Halomonas and Shewanella was significantly increased in patients with sexually transmitted HIV-infection compared with healthy controls. Alterations of the gut microbiota during HIV infection were also indicated to be associated with the route of HIV transmission. Certain bacteria may be potential biomarkers for HIV infection in patients from Guangzhou, China.

Gut microbiota mediates the effects of inulin on enhancing sulfomucin production and mucosal barrier function in a pig model.

Dietary fibers (DFs) have many beneficial effects on intestinal health by ameliorating intestinal inflammation and modulating the microbial community composition, thereby affecting the barrier function. This study aims to characterize the gut microbiota of pigs fed with DFs, revealing a link between the intestinal microbiota and mucin chemotypes. Pigs (six per group) were randomly allotted to consume one of the following diets: control (CON) or a diet supplemented with 5% microcrystalline cellulose (MCC) or inulin (INU) for 72 days. We found that INU but not MCC enhanced the colonic barrier function by promoting the expression of ZO-1, Occludin and MUC2 and reducing the colonic crypt depth. INU increased sulfomucin production and mRNA levels of sulfotransferases Gal3ST1 and Gal3ST2. Goblet cells in the ileum were found to contain predominantly sialomucins while colonic goblet cells were dominated by sulfomucins with sialomucins absent. DF consumption increased the concentrations of short-chain fatty acids (SCFAs) of the ileum and colon compared to the CON diet. Moreover, the results of 16S rRNA gene sequencing analysis revealed that DFs significantly altered the composition of ileal and colonic mucosal microbiota. Network analysis indicated that INU-induced changes in bacterial genera and SCFAs, such as Akkermansia and butyrate, were significantly related with sulfomucins and the mucosal barrier function-gene in pigs. Collectively, these findings suggest that the intestinal mucosal microbiota and SCFAs induced by INU play a crucial role in modulating the chemotypes of mucin and the barrier function.

Intestinal mucosal bacterial diversity of antibiotic-associated diarrhea (AAD) mice treated with Debaryomyces hansenii and Qiweibaizhu powder

The aim was to investigate the combined effect of Debaryomyces hansenii and Qiweibaizhu powder (QWBZP) on the bacterial diversity of the intestinal mucosa of antibiotic-associated diarrhea (AAD) mice, for the potential treatment of diarrhea, especially which is induced by administration of antibiotics. Eighteen (18) mice were randomly assigned to three equal groups of six mice, namely Normal (mn group), Placebo control (mm group) and D. hansenii and QWBZP (DQ) treatment (mdq group). Mice were gavaged with a solution (23.33mL·kg-1·day-1) consisting of gentamicin and cefradine to establish AAD. The DQ treatment group was gavaged with DQ for 4days, and sterile water was used as a placebo control. The metagenome DNA of the intestinal mucosal microbiota was extracted, and the 16S rRNA gene was sequenced. Analysis showed that there were 288 OTUs for the normal group, 443 for the placebo control group, and 229 for the DQ treatment group. Phylogenetically, the gut microbiota of the DQ treatment group and the normal group were closer to each other than to the placebo control group. Both the DQ and placebo-treated groups included Stenotrophomonas, Robinsoniella, Bacteroidales S24-7 group norank, Citrobacter, and Glutamicibacter, but their abundances were significantly higher in the DQ treatment group than in the placebo control group. This suggested that the combined use of D. hansenii and QWBZP overcame the influence of dysbacteriosis and could lead to the recovery of intestinal mucosal microbiota homeostasis. This positive effect is likely related to short-chain fatty acid (SCFA)-producing bacteria, such as members of Micrococcaceae, Lachnospiraceae, and Bacteroidales S24-7 group, which could play beneficial roles in protecting the mucosal barrier and stimulating the immune response in mice.

Microbiotal characteristics colonized in intestinal mucosa of mice with diarrhoea and repeated stress

To understand the role of intestinal mucosal microbiota on mental stress-related diarrhoea, we collected the intestinal mucosa of mice treated with Folium senna extract gavage combined with restraint and tail pinch stress for 7days; and intestinal mucosal microbiota characteristics were analyzed by 16S rRNA Pacbio SMRT gene full-length sequencing. The results showed that the diversity (i.e., alpha diversity including the Chao1, Simpson, ACE, and Shannon indices and beta diversity including the NMDS of weighted UniFrac distances) and composition of the microbial community in the intestinal mucosa of mice with diarrhoea and repeated stress changed significantly (P < 0.05). In the co-occurrence network, Staphylococcus sciuri and Escherichia fergusonii was identified as putative keystone species. Moreover, the characteristics of the intestinal microbial species was analyzed by LEfSe, Metastats, and group difference, and ten altered gut microbiota species can be used as characteristic microbes in the mice with diarrhoea and repeated stress: the abundances of Stigmatella aurantiaca, Candidatus arthromitus sp. SFB-mouse, Erythrobacter gaetbuli, Desulfitobacterium hafniense, Ochrobactrum pituitosum, and Candidatus arthromitus sp. SFB-mouse-NL in the model group were significantly lower than those in the control group (P < 0.05); whereas Microbacterium dextranolyticum, Klebsiella pneumoniae, Escherichia sp. BBDP27, and Streptococcus danieliae were enriched in the control group (P < 0.05). Collectively, mental stress-related diarrhoea increased the intestinal microbiota diversity. The species associated with mental stress-related diarrhoea including Microbacterium dextranolyticum, Klebsiella pneumoniae, Escherichia sp. BBDP27, and Streptococcus danieliae were significantly enriched; while the species which are beneficial to mental stress-related diarrhoea are Stigmatella aurantiaca, Candidatus arthromitus sp. SFB-mouse, Erythrobacter gaetbuli, Desulfitobacterium hafniense, Ochrobactrum pituitosum, and Candidatus arthromitus sp. SFB-mouse-NL for its significantly depleted.