The impact of altered intestinal microbiota on intestinal immune function after acute exhaustive exercise in mice

Results of the First Pilot Randomized Controlled Trial of Fecal Microbiota Transplant In Pediatric Ulcerative Colitis: Lessons, Limitations, and Future Prospects

Most studies have reported fecal microbiota transplantation (FMT) as an effective secondary option for Crohn's disease (CD). However, there is little data on FMT as a first-line treatment for CD. In our study we explore the rates of clinical and endoscopic remission and mucosal healing after FMT plus partial enteral nutrition (PEN), as a first-line treatment for active CD in children. We retrospectively enrolled pediatric CD patients who underwent PEN or PEN plus FMT treatment at diagnosis from November 2016 to July 2019 at the Pediatric Department, Tongji Hospital. The two groups were defined as FMT group (repeated and multiple doses of FMT plus PEN) or PEN group (PEN alone). All the patients received PEN intervention. At baseline and week 8- 10, the FMT group was administered multiple doses of FMT to help induce and maintain remission. All patients were evaluated at week 8- 10 and 18-22 via clinical and relevant laboratory parameters and endoscopic results. The clinical and endoscopic remission and mucosal healing rates were compared between the two groups at different time points after the therapy. Twenty-five newly diagnosed active CD patients were included in the study, containing 7 females and 18 males with a median age of 11. 1 ± 2.3 years. 13 and 12 patients were assigned to the PEN and FMT groups, respectively. At week 8-10, clinical remission was obtained in 83.3% and 53.8% of the FMT and PEN groups, respectively (p=0.202). The endoscopic remission rates were 72.7% for FMT and 25.0% for PEN (p=0.039), whereas the mucosal healing rates were 27.2% for FMT and 0% for PEN (p=0.093). At week 18-22, clinical remission was achieved in 72.7% and 20.0% of patients in the FMT and PEN groups, respectively (p=0.03). Theendoscopic remission rates were 66.6% and 12.5% in the FMT and PEN groups, respectively (p=0.05), whereas the mucosal healing rates were 55.5% and 0% in FMT and PEN groups, respectively (p=0.029). This study demonstrate that FMT plus PEN can be used as a first-line treatment for active CD in children.

Fecal microbiota transplantation revealed the function of folic acid on reducing abdominal fat deposition in broiler chickens mediated by gut microbiota

Simple SummaryAntibiotic resistance is a major concern, and dogs with tylosin-responsive enteropathy (TRE) often receive the antibiotic for extended periods. Fecal microbiota transplantation (FMT) could be an alternative treatment. This clinical trial compared the impact of oral FMT/placebo capsules on 14 TRE dogs (7 FMT, 7 placebo) by analyzing their fecal consistencies and gut microbiomes. The microbial engraftment was assessed based on the gut microbiome of the single stool donor. In addition, the gut microbiome of these 14 dogs was compared with the fecal microbial composition of 30 healthy dogs. During the 4-week treatment trial, 5/7 dogs in the FMT group (71.4%) and 3/6 dogs in the placebo group (50%) did not relapse. After the trial treatment and 4-week follow-up, 2 more dogs in the FMT group and 1 dog in the placebo group relapsed, but none in the placebo group. The differences in relapse rates between the FMT and placebo groups were not statistically significant. TRE dogs had lower microbiome diversity when on tylosin, but it increased after treatment with FMT or placebo. On average, 30.4% of the donor bacterial strains were engrafted into FMT recipients. Overall, the FMT efficacy was slightly higher than the placebo, but the difference was not statistically significant, possibly because of the small sample size of the study. Future clinical trials should include larger sample sizes to determine the efficacy of oral FMT in dogs with TRE or chronic enteropathies.A clinical trial was conducted to evaluate the effect of fecal microbiota transplantation (FMT) on the canine chronic enteropathy clinical activity index (CCECAI), fecal consistency, and microbiome of dogs with tylosin-responsive enteropathy (TRE). The trial consisted of four phases: (1) screening with discontinuation of tylosin for 4 weeks, (2) inclusion with re-introduction of tylosin for 3–7 days, (3) treatment with FMT/placebo for 4 weeks, and (4) post-treatment with follow-up for 4 weeks after treatment cessation. The study found that the treatment efficacy of FMT (71.4%) was slightly higher than that of placebo (50%), but this difference was not statistically significant due to underpowering. The most abundant bacterial species detected in the fecal microbiomes of dogs with TRE before FMT or placebo treatment were Blautia hansenii, Ruminococcus gnavus, Escherichia coli, Clostridium dakarense, Clostridium perfringens, Bacteroides vulgatus, and Faecalimonas umbilicata. After FMT, the microbiomes exhibited increases in Clostridium dakarense, Clostridium paraputrificum, and Butyricicoccus pullicaecorum. The microbiome alpha diversity of TRE dogs was lower when on tylosin treatment compared to healthy dogs, but it increased after treatment in both the FMT and placebo groups. Comparisons with the stool donor showed that, on average, 30.4% of donor strains were engrafted in FMT recipients, with the most common strains being several Blautia sp., Ruminococcus gnavus, unclassified Lachnoclostridium, Collinsella intestinalis, and Fournierella massiliensis.

To explore the effects of different transplantation frequencies and time of fecal microbiota transplantation on mice. Twenty-four C57BL/6J mice were randomly divided into control group, fecal microbiota transplantation group 1(FMT1), fecal microbiota transplantation group 2(FMT2), and fecal microbiota transplantation group 3(FMT3). The control group was used as the donor of fecal microbiota transplantation, and the FMT1, FMT2, and FMT3 groups were intervened with mixed antibiotics(200 μL/d) for 2 weeks, and received fecal bacterial suspension(200 μL/d). The transplantation time of the FMT1 group frequency was 1 time/d for 1 weeks, the FMT2 group was 1 time/d for 2 weeks, and the FMT3 group was 3 times/week for 2 weeks. At the end of the experiment, the feces of the mice were collected to analyze the gut microbiota. Compared with the control group, there were more independent Amplicon Sequence Variants in the intestinal microbiota of mice in FMT1 group, FMT2 group and FMT3 group, and the ACE index and Chao1 index were significantly reduced(P<0.05). Beta diversity showed differences between fecal microbiota transplantation and control groups, with FMT2 and control groups being the closest. At the phylum level, there were two species in FMT1 group and one species in FMT3 group showed statistically significant differences compared with control group(P<0.05). However, there was no significant difference between the FMT2 group and the control group. At the genus level, there were 6 species in the FMT1 with statistically significant differences from the control group(P<0.05), and 2 species in the FMT2, 5 species in the FMT3 respectively. Among which FMT2 group has the least number of species that differed from the control group, suggesting that the compitsition of its intestinal microbiota is closet to that of the control group. Fecal bacteria transplantation helps to restore the intestinal microbiota structure of mice cleaned by antibiotics, and different transplantation frequencies and transplantation times have different recovery effects on the intestinal microbiota of mice pretreated with antibiotics, and the fecal bacteria transplantation effect is better with 1 time/d lasting 2 weeks.

Aims: We investigated whether faecal microbiota transplantation (FMT) decreases intra-abdominal pressure (IAP) and improves gastrointestinal (GI) dysfunction and infectious complications in acute pancreatitis (AP).Methods: In this first randomised, single-blind, parallel-group, controlled study, we recruited and enrolled consecutive patients with AP complicated with GI dysfunction. Eligible participants were randomly assigned to receive faecal transplant (n = 30) or normal saline (n = 30) via a nasoduodenal tube once and then again 2 days later. The primary endpoint was the rate of IAP decline; secondary endpoints were GI function, infectious complications, organ failure, hospital stay and mortality. Analyses were based on intention to treat.Results: We enrolled 60 participants and randomly assigned them to the FMT (n = 30) or control (n = 30) group. Baseline characteristics and disease severity were similar for both groups. IAP decreased significantly 1 week after intervention in both groups, with no difference in the IAP decline rate between FMT and Control group [0.1 (−0.6, 0.5) vs. 0.2 (−0.2, 0.6); P = 0.27]. Normal gastrointestinal failure (GIF) scores were achieved in 12 (40%) patients in the FMT group and 14 (47%) in the control group, with no significant difference (P = 0.60). However, D-lactate was significantly elevated in the FMT group compared to the control group, as calculated by the rate of decline [−0.3 (−3.7, 0.8) vs. 0.4 (−1.1, 0.9); P = 0.01]. Infectious complications occurred in 15 (50%) and 16 (53.33%) patients in the FMT and control groups, respectively (P = 0.80). However, interleukin-6 (IL-6) was significantly elevated in the FMT group compared to the control group, as calculated by the rate of decline [0.4 (−3.6, 0.9) vs. 0.8 (−1.7, 1.0); P = 0.03]. One participant experienced transient nausea immediately after FMT, but no serious adverse events were attributed to FMT.Conclusion: FMT had no obvious effect on IAP and infectious complications in AP patients, though GI barrier indictors might be adversely affected. Further multi-centre studies are needed to confirm our findings. The study was registered at https://clinicaltrials.gov (NCT02318134).

Objective To observe the effects of fecal microbiota transplantation (FMT) on acute lung injury (ALI) induced by lipopolysaccharide (LPS) in rats and to investigate the possible mechanism in order to provide new thoughts for the treatment of acute lung injury. Methods Forty-five adult healthy male SPF level SD rats were randomly divided into three groups of normal saline (NS), LPS and FMT with 15 in each group. ALI was induced by intraperitoneal injection of rats with 5 mg/kg of LPS. The FMT group was given 10 ml/kg of fecal microbiota solution intervention (twice a day for two consecutive days) after ALI induction. Five rats in each group were randomly selected and sacrificed at 24 h, 48 h and 72 h after intervention. Pathological changes in lung tissues were examined with hematoxylin and eosin (HE) staining and pathological scores were assessed. Right lung samples were weighed to measure wet/dry (W/D) ratios. Abdominal aorta blood samples were collected for PaO2 analysis. The levels of tumor necrosis factor-α (TNF-α), interleukin-1β (IL-1β) and interleukin 6 (IL-6) in bronchoalveolar lavage fluid (BALF) were detected by enzyme linked immunosorbent assay (ELISA). Expression of transforming growth factor-β (TGF-β) and intracellular signal transduction protein Smads (Smad3 and Smad7) at mRNA level was analyzed by reverse transcription-polymerase chain reaction (RT-PCR). Expression of ERK1/2 and p-ERK1/2 at protein level was detected by Western blot (WB). Rat fecal samples were collected to extract DNA and the PCR products of V3 and V4 regions were sequenced by Illumina MiSeq. Bioinformatics analysis on microbiota was conducted with Illumina MiSeq based on operational taxonomic unit (OTU) clustering. Results Compared with the NS group, the LPS group showed significantly widened alveolar septa, massive inflammatory cell infiltration and some alveolar collapse in lung tissues. Compared with the LPS group, the FMT group showed significantly alleviated inflammatory cell infiltration, reduced swelling in pulmonary interstitial tissues, and less damage to pulmonary structure. Compared with the NS group, the W/D lung ratio, PaO2 concentration and TNF-α, IL-1 and IL-6 levels in BALF in the LPS group significantly increased at 24 h after intervention and then gradually decreased, but still higher than those in the NS group (P<0.05). The W/D lung ratio, PaO2 concentration and TNF-α, IL-1 and IL-6 levels in BALF of the FMT group were significantly lower than those of the LPS group (P<0.05). Compared with the NS group, the expression of TGF-β1 and Smad3 at mRNA level in lung tissues increased at all time points in the LPS group, while the expression of Smad7 at mRNA level decreased (P<0.05). The expression of TGF-β1 and Smad3 at mRNA level in the FMT group were significantly lower than that in the LPS group (P<0.05), while the expression of Smad7 at mRNA level was higher (P<0.05). Compared with the NS group, p-ERK expression significantly increased in the LPS group with the highest expression at 24 h and gradually decreased at 48 h and 72 h (P<0.05). The expression of p-ERK in the FMT group was significantly lower than that in the LPS group (P<0.05). Results of the intestinal microbiota sequencing revealed 1 362 OTU in the NS group, 1 443 OTU in the LPS group and 1 510 OTU in the FMT group. There were 336 OTU shared by them accounting for 26.4%. The constitution of intestinal microbiota in the LPS group was significantly different with that of normal rats and characterized by high diversity with increased Firmicutes/Bacteroidetes ratio and decreased gene abundance of intestinal microbiota. After the intervention of fecal bacteria transplantation, the constitution of intestinal microbiota in the FMT group was similar to that of the NS group, showing decreased Firmicutes/Bacteroidetes ratio and increased gene abundance of the bacterial community (P<0.05). Conclusions FMT might inhibit immune inflammation, reduce the production and secretion of inflammatory markers in the body, alleviate alveolar epithelial damage and abnormal repair through regulating intestinal microbiota and TGF-β1/Smads/ERK pathway to improve the LPS-induced endotoxic ALI in rats. Key words: Acute lung injury; Fecal microbiota transplantation; High-throughput Sequencing; Lipopolysaccharide

Gut microbiota (GM) dysbiosis is closely related to several metabolic diseases such as hypertension, obesity, and Alzheimer's disease. However, little is known about the causal relationship between GM dysbiosis and osteoporosis. In our work, 32 3-month-old female SD rats were randomly divided into two groups: the fecal microbiota transplantation (FMT) group and the control group. The supernatant of feces from senile osteoporotic rats was transplanted to the FMT group and the same amount of sterile saline was given to the control rats. After 12 and 24weeks, all rats were sacrificed, and the serum, bone, fecal feces, and intestine tissue were collected for the subsequent analysis. The osteocalcin (OC), CTX, and P1NP of the FMT group increased significantly at 12 and 24weeks compared with the control group (P < 0.05). Furthermore, the BV, BV/TV, Tb.N, and Tb.Th decreased significantly in the FMT group (P < 0.05). The alpha diversity (ACE, Chao) of the FMT group was higher than the control at 24weeks (P < 0.05). The beta diversity was close between the FMT rats and the donor rats. In addition, GM from donor rats changed the GM composition and function of the FMT rats, which was similar to that of the donor rats at 24weeks. The impaired intestinal structure and the decreased expression of occludin, claudin, and ZO-1 were found in FMT rats. In conclusion, GM dysbiosis by transferring the feces from senile osteoporotic rats to young rats could induce osteoporosis. The changed GM and the impaired intestinal barrier contributed to the pathogenesis of osteoporosis.

Lyophilised oral faecal microbiota transplantation for ulcerative colitis (LOTUS): a randomised, double-blind, placebo-controlled trial

Faecal microbiota transplantation for first or second Clostridioides difficile infection (EarlyFMT): a randomised, double-blind, placebo-controlled trial

目的探讨粪菌移植（FMT）治疗糖皮质激素耐药胃肠道急性移植物抗宿主病（GI-aGVHD）的临床疗效。方法纳入2017年3月至2022年3月期间在徐州医科大学附属淮安医院行异基因造血干细胞移植（allo-HSCT）后发生糖皮质激素耐药GI-aGVHD的29例血液病患者，其中19例行FMT治疗（FMT组），10例未接受FMT治疗（对照组）。观察疗效及安全性，分析FMT治疗前后肠道菌群丰度、淋巴细胞亚群比例、外周血炎症因子及GVHD生物标志物的变化。结果①13例患者（68.4％）在FMT后临床症状达到完全缓解，对照组2例患者（20.0％）达到完全缓解，差异有统计学意义（P<0.05）。FMT后肠道微生物群多样性增加，并逐渐恢复至正常水平，未发生FMT相关感染。②与对照组比较，FMT组治疗后CD3+、CD8+细胞占比降低，CD4+、调节性T细胞（Treg）及CD4+/CD8+细胞比值升高（P值均<0.05），IL-6浓度低于对照组［4.15（1.91～5.71）ng/L对6.82（2.40～8.91）ng/L，P＝0.040］，IL-10浓度高于对照组［12.11（5.69～20.36）ng/L对7.51（4.10～9.58）ng/L，P＝0.024］。胰岛衍生蛋白3α（REG3α）在发生GI-aGVHD时明显升高，FMT组治疗后REG3α水平低于对照组［30.70（10.50～105.00）µg/L对74.35（33.50～139.50）µg/L，P＝0.021］。结论FMT是糖皮质激素耐药GI-aGVHD的有效、安全治疗方法。FMT可促进肠道菌群多样性的恢复、调节炎症因子和上调Treg细胞表达。

Fecal microbiota transplantation (FMT) could decolonize multidrug-resistant organisms. We investigated FMT effectiveness and safety in the eradication of carbapenem-resistant Enterobacteriaceae (CRE) and vancomycin-resistant enterococci (VRE) intestinal colonization. A prospective non-randomized comparative study was performed with 48 patients. FMT material (60 g) was obtained from a healthy donor, frozen, and administered via endoscopy. The primary endpoint was 1-month decolonization, and secondary endpoints were 3-month decolonization and adverse events. Microbiota analysis of fecal samples was performed using 16S rRNA sequencing. Intention-to-treat analysis revealed overall negative conversion between the FMT and control groups at 1 (26% vs. 10%, p = 0.264) and 3 (52% vs. 24%, p = 0.049) months. The 1-month and 3-month CRE clearance did not differ significantly by group (36% vs. 10%, p = 0.341; and 71% vs. 30%, p = 0.095, respectively). Among patients with VRE, FMT was ineffective for 1-month or 3-month negative conversion (13% vs. 9%, p > 0.999; and 36% vs. 18%, p = 0.658, respectively) However, cumulative overall negative-conversion rate was significantly higher in the FMT group (p = 0.037). Enterococcus abundance in patients with VRE significantly decreased following FMT. FMT may be effective at decolonizing multidrug-resistant organisms in the intestinal tract.

BackgroundThere is intense interest about whether modulating gut microbiota can impact systemic metabolism. We investigated the safety of weekly oral fecal microbiota transplantation (FMT) capsules from healthy lean donors and their ability to alter gut microbiota and improve metabolic outcomes in patients with obesity.Methods and findingsFMT-TRIM was a 12-week double-blind randomized placebo-controlled pilot trial of oral FMT capsules performed at a single US academic medical center. Between August 2016 and April 2018, we randomized 24 adults with obesity and mild–moderate insulin resistance (homeostatic model assessment of insulin resistance [HOMA-IR] between 2.0 and 8.0) to weekly healthy lean donor FMT versus placebo capsules for 6 weeks. The primary outcome, assessed by intention to treat, was change in insulin sensitivity between 0 and 6 weeks as measured by hyperinsulinemic euglycemic clamps. Additional metabolic parameters were evaluated at 0, 6, and 12 weeks, including HbA1c, body weight, body composition by dual-energy X-ray absorptiometry, and resting energy expenditure by indirect calorimetry. Fecal samples were serially collected and evaluated via 16S V4 rRNA sequencing. Our study population was 71% female, with an average baseline BMI of 38.8 ± 6.7 kg/m2 and 41.3 ± 5.1 kg/m2 in the FMT and placebo groups, respectively. There were no statistically significant improvements in insulin sensitivity in the FMT group compared to the placebo group (+5% ± 12% in FMT group versus −3% ± 32% in placebo group, mean difference 9%, 95% CI −5% to 28%, p = 0.16). There were no statistically significant differences between groups for most of the other secondary metabolic outcomes, including HOMA-IR (mean difference 0.2, 95% CI −0.9 to 0.9, p = 0.96) and body composition (lean mass mean difference −0.1 kg, 95% CI −1.9 to 1.6 kg, p = 0.87; fat mass mean difference 1.2 kg, 95% CI −0.6 to 3.0 kg, p = 0.18), over the 12-week study. We observed variable engraftment of donor bacterial groups among FMT recipients, which persisted throughout the 12-week study. There were no significant differences in adverse events (AEs) (10 versus 5, p = 0.09), and no serious AEs related to FMT. Limitations of this pilot study are the small sample size, inclusion of participants with relatively mild insulin resistance, and lack of concurrent dietary intervention.ConclusionsWeekly administration of FMT capsules in adults with obesity results in gut microbiota engraftment in most recipients for at least 12 weeks. Despite engraftment, we did not observe clinically significant metabolic effects during the study.Trial registrationClinicalTrials.gov NCT02530385.

BackgroundThe global population of obese individuals is increasing, affecting human health. High-fat diets are a leading cause of this epidemic, and animal models, such as mice, are often used in related research. Obese individuals have a different gut microbiota composition from non-obese ones, characterized by a sizeable population of certain bacteria associated with fat storage. The gut microbiome plays a significant role in regulating human physiological and metabolic functions. Links between obesity, high-fat diets and gut microbiota have become hot topics of discussion. Recently, research on the modulation of the gut microbiota has focused on fecal microbiota transplantation (FMT), which has been recognized as an effective method of studying the function of gut microbiota.ObjectivesThe purpose of this study was to investigate how the gut microbiota of Suncus murinus, a naturally obesity-resistant animal, through FMT, affected the ecology of the gut microbiota of high-fat diet induced obese mice.MethodsIn this study, Suncus murinus was used as a donor for FMT. High-fat diet induced C57BL/6NCrSIc mice were used as recipients, the body weight changes were measured and changes in their gut flora were analyzed using a 16S rRNA gene analysis.ResultsThe study found that, after the FMT procedure, the FMT group tended to have a lower body weight than the control group. At the phylum level, the most predominant phyla in all groups were Firmicutes and Proteobacteria, while Deferribacteres was not detected in the FMT or antibiotic administration groups, and Bacteroidetes was not present in the antibiotic administration group. At the genus level, the FMT group had significantly lower OTU richness than the control group but greater diversity than the control group.ConclusionsThese results indicate that FMT from Suncus murinus can help reorganize and improve the gut microbiota of mice in a balanced and diverse ecosystem.

Fecal microbiota transplantation (FMT) is an emerging and effective therapy for the treatment of inflammatory bowel disease (IBD). Previous studies have reported that compared with FMT, whole intestinal microbiota transplantation (WIMT) can more precisely replicate the community structure and reduce the inflammatory response of the host. However, it remains unclear whether WIMT is more effective in alleviating IBD. To examine the efficacy of WIMT and FMT in the intervention of IBD, GF (Germ-free) BALB/c mice were pre-colonized with whole intestinal microbiota or fecal microbiota before being treated with dextran sodium sulfate (DSS). As expected, the symptoms of colitis were alleviated by both WIMT and FMT, as demonstrated by the prevention of body weight loss and decreased the Disease activity index and histological scores in mice. However, WIMT's anti-inflammatory effect was superior to that of FMT. In addition, the inflammatory markers myeloperoxidase (MPO) and eosinophil peroxidase were dramatically downregulated by WIMT and FMT. Furthermore, the use of two different types of donors facilitated the regulation of cytokine homeostasis in colitis mice; the level of the pro-inflammatory cytokine IL-1β in the WIMT group was significantly lower than that in the FMT group, while the level of the anti-inflammatory factor IL-10 was significantly higher than that in the FMT group. Both groups showed enhanced expression of occludin to protect the intestinal barrier in comparison with the DSS group, and the WIMT group demonstrated considerably increased levels of ZO-1. The sequencing results showed that the WIMT group was highly enriched in Bifidobacterium, whereas the FMT group was significantly enriched in Lactobacillus and Ochrobactrum. Correlation analysis revealed that Bifidobacterium was negatively correlated with TNF-α, whereas Ochrobactrum was positively correlated with MPO and negatively correlated with IL-10, which might be related to different efficacies. Functional prediction using PICRUSt2 revealed that the FMT group was considerably enriched in the L-arginine biosynthesis I and L-arginine biosynthesis IV pathway, whereas the WIMT group was enriched in the L-lysine fermentation to acetate and butanoate pathway. In conclusion, the symptoms of colitis were subsided to varying degrees by the two different types of donors, with the WIMT group being more effective than the FMT group. This study provides new information on clinical interventions for IBD.