Faecalibacterium Research Articles

Synergy and oxygen adaptation for development of next-generation probiotics

The human gut microbiota has gained interest as an environmental factor that may contribute to health or disease1. The development of next-generation probiotics is a promising strategy to modulate the gut microbiota and improve human health; however, several key candidate next-generation probiotics are strictly anaerobic2 and may require synergy with other bacteria for optimal growth. Faecalibacterium prausnitzii is a highly prevalent and abundant human gut bacterium associated with human health, but it has not yet been developed into probiotic formulations2. Here we describe the co-isolation of F. prausnitzii and Desulfovibrio piger, a sulfate-reducing bacterium, and their cross-feeding for growth and butyrate production. To produce a next-generation probiotic formulation, we adapted F. prausnitzii to tolerate oxygen exposure, and, in proof-of-concept studies, we demonstrate that the symbiotic product is tolerated by mice and humans (ClinicalTrials.gov identifier: NCT03728868) and is detected in the human gut in a subset of study participants. Our study describes a technology for the production of next-generation probiotics based on the adaptation of strictly anaerobic bacteria to tolerate oxygen exposures without a reduction in potential beneficial properties. Our technology may be used for the development of other strictly anaerobic strains as next-generation probiotics.

Effect of Benaglutide on Gut Microbiota and Fecal Metabolites in Patients with Type 2 Diabetes Mellitus.

Benaglutide is a glucagon-like peptide-1 receptor agonist (GLP-1RA) that has been approved in the treatment of type 2 diabetes mellitus (T2DM). It is known to lead to significant weight loss, and it is hypothesized that changes in gut microbiota may play a significant role in such weight loss. However, it is unclear how gut microbiota and metabolites change as a result of benaglutide treatment. Healthy participants and patients with T2DM were included in this study. They received differentiated treatments, and stool specimens were collected separately. These stool specimens were subjected to 16S ribosomal RNA amplicon and metagenomic sequencing to create fecal metabolomic profiles. The diversity of gut microbiota and metabolic products in the stools of each participant was analyzed. The data showed that Faecalibacterium prausnitzii was abundant in the gut microbiota of the control group, which was entirely made up of healthy individuals; however, it showed a statistically significant decrease in patients with T2DM treated with metformin alone, while no significant decrease was observed in patients treated with metformin combined with benaglutide. A metagenomic analysis revealed that benaglutide could improve the fecal microbiota diversity in patients with T2DM. Furthermore, there was a statistically significant correlation between the changes in the metabolites of patients with T2DM and the changes in their gut microbiota (including F. prausnitzii) after treatment with metformin and benaglutide. These findings suggest that the weight-reducing effect of benaglutide is attributed to its ability to normalize the gut microbiota of patients with T2DM, particularly by increasing the abundance of F. prausnitzii.

Carnobacterium maltaromaticum boosts intestinal vitamin D production to suppress colorectal cancer in female mice.

Carnobacterium maltaromaticum was found to be specifically depleted in female patients with colorectal cancer (CRC). Administration of C.maltaromaticum reduces intestinal tumor formation in two murine CRC models in a female-specific manner. Estrogen increases the attachment and colonization of C.maltaromaticum via increasing the colonic expression of SLC3A2 that binds to DD-CPase of this bacterium. Metabolomic and transcriptomicprofiling unveils the increased gut abundance of vitamin D-related metabolites and the mucosal activation of vitamin D receptor (VDR) signaling in C.maltaromaticum-gavaged mice in a gut microbiome- and VDR-dependent manner. Invitro fermentation system confirms the metabolic cross-feeding ofC.maltaromaticum with Faecalibacterium prausnitzii to convert C.maltaromaticum-produced 7-dehydrocholesterol into vitamin D for activating the host VDR signaling. Overall, C.maltaromaticum colonizes the gut in an estrogen-dependent manner and acts along with other microbes to augment the intestinal vitamin D production to activate the host VDR for suppressing CRC.

Integrated analysis of the transcriptome and its interaction with the metabolome in metabolic associated fatty liver disease: Gut microbiome signatures, correlation networks, and effect of PNPLA3 genotype.

Interactions between communities of the gut microbiome and with the host could affect the onset and progression of metabolic associated fatty liver disease (MAFLD), and can be useful as new diagnostic and prognostic biomarkers. In this study, we performed a multi-omics approach to unravel gut microbiome signatures from 32 biopsy-proven patients (10 simple steatosis -SS- and 22 steatohepatitis -SH-) and 19 healthy volunteers (HV). Human and microbial transcripts were differentially identified between groups (MAFLD vs. HV/SH vs. SS), and analyzed for weighted correlation networks together with previously detected metabolites from the same set of samples. We observed that expression of Desulfobacteraceae bacterium, methanogenic archaea, Mushu phage, opportunistic pathogenic fungi Fusarium proliferatum and Candida sorbophila, protozoa Blastocystis spp. and Fonticula alba were upregulated in MAFLD and SH. Desulfobacteraceae bacterium and Mushu phage were hub species in the onset of MAFLD, whereas the activity of Fonticula alba, Faecalibacterium prausnitzii, and Mushu phage act as key regulators of the progression to SH. A combination of clinical, metabolomic, and transcriptomic parameters showed the highest predictive capacity for MAFLD and SH (AUC=0.96). In conclusion, faecal microbiome markers from several community members contribute to the switch in signatures characteristic of MAFLD and its progression towards SH.

Faecalibacterium prausnitzii alleviates inflammatory arthritis and regulates IL-17 production, short chain fatty acids, and the intestinal microbial flora in experimental mouse model for rheumatoid arthritis

BackgroundRheumatoid arthritis (RA) is a systemic chronic inflammatory disease that leads to joint destruction and functional disability due to the targeting of self-antigens present in the synovium, cartilage, and bone. RA is caused by a number of complex factors, including genetics, environment, dietary habits, and altered intestinal microbial flora. Microorganisms in the gut bind to nod-like receptors and Toll-like receptors to regulate the immune system and produce various metabolites, such as short-chain fatty acids (SCFAs) that interact directly with the host. Faecalibacterium prausnitzii is a representative bacterium that produces butyrate, a well-known immunomodulatory agent in the body, and this microbe exerts anti-inflammatory effects in autoimmune diseases.MethodsIn this study, F. prausnitzii was administered in a mouse model of RA, to investigate RA pathology and changes in the intestinal microbial flora. Using collagen-induced arthritic mice, which is a representative animal model of RA, we administered F. prausnitzii orally for 7 weeks.ResultsThe arthritis score and joint tissue damage were decreased in the mice administered F. prausnitzii compared with the vehicle-treated group. In addition, administration of F. prausnitzii reduced the abundance of systemic immune cells that secrete the pro-inflammatory cytokine IL-17 and induced changes in SCFA concentrations and the intestinal microbial flora composition. It also resulted in decreased lactate and acetate concentrations, an increased butyrate concentration, and altered compositions of bacteria known to exacerbate or improve RA.ConclusionThese results suggest that F. prausnitzii exerts a therapeutic effect on RA by regulation of IL-17 producing cells. In addition, F. prausnitzii modify the microbial flora composition and short chain fatty acids in experimental RA mouse model.

Characteristics of the microbiome in patients with symptomatic uncomplicated colonic diverticular disease in the central region of Ukraine

The aim of the study was to analyze the age structure, correlation with body mass index in patients with symptomatic uncomplicated diverticular disease (SUDD) in the central region of Ukraine. To reveal the prevalence of left-sided, right-sided or total diverticular lesions of the large intestine, to evaluate the microbiota composition, to identify peculiarities of the enterotype. Materials and methods. The study included 125 patients with SUDD aged 18 to 90 years who were at a Gastroenterology Hospital in the period from 2019 to 2022. All the patients were residents of the central region of Ukraine. Body mass index was calculated for all patients, total colonoscopy was performed, and fecal analysis was done using culturing and qRT-PCR methods. Results. The incidence of SUDD among patients in the central region of Ukraine increases with age and reaches its peak in the 71–80 age group. Almost all age groups are dominated by women. Left-sided localization of diverticula occurs in 84.8 % of patients, right-sided in 8.0 %, total in 7.2 %. The majority of patients with diverticular disease (87.2 %) are overweight or with varying degrees of obesity. Patients with SUDD are characterized by a decrease in the main representatives of obligate microflora, namely, Bifidobacterium, Lactobacillus, Escherichia coli. In 46.4 % of patients, representatives of opportunistic flora are detected, in 24.0 % – an increase in fungi from the genus Candida. Also, changes in enterotypes are detected, namely an increase in Firmicutes and Actinobacteria, with a decrease in Bacteroides. The number of Faecalibacterium prausnitzii tends to decrease, while the Akkermansia muciniphila count is unchanged. Conclusions. The incidence of SUDD increases with age among the population of the central region of Ukraine. Among patients with SUDD, women predominate in almost all age groups. 87.2 % of patients with SUDD are overweight or with various degrees of obesity. The left-sided localization of diverticula prevails among patients with SUDD in the central region of Ukraine – 84.8 %. Patients with SUDD in the central region of Ukraine are characterized by the decrease in the main representatives of obligate microflora, namely: Bifidobacterium, Lactobacillus, Escherichia coli. In 46.4 % of patients with SUDD, representatives of conditionally pathogenic flora are found, in 24.0 % – the increase in fungi from the genus Candida. Changes in enterotypes are detected, namely the increase in Firmicutes and Actinobacteria, with the decrease in Bacteroides. Faecalibacterium prausnitzii tends to decrease, while Akkermansia muciniphila are unchanged.

Resistance to antimicrobial preparations of Enterococcus isolated from milk during infectious mastitis in cows

For successful pharmacotherapy of animals, it is important to identify species of Enterococcus faecalis pathogens isolated from milk in infectious mastitis of cows, analyze the antibiogram and interpret the results, and laboratory control over the mechanisms of antibiotic resistance. This is necessary for veterinarians to rationally select antibacterial drugs and predict their clinical effectiveness. The purpose of the study was to study the resistance to antimicrobial drugs of Enterococcus faecalis bacteria isolated from milk in infectious mastitis of cows. In the period from 2021 to 2022, 100 strains of microorganisms were isolated from the mastitis milk of cows. The obtained strains were identified as gram-negative bacteria in 32% of isolation cases, gram-positive bacteria - 68%, of which 14% were Enterococcus faecalis. When interpreting the antibiogram, most isolates of Enterococcus faecalis are sensitive to tetracycline in 21.4% of isolation cases, ampicillin, amoxicillin in 100%. It was noted resistant to tetracycline in 78.6% of cases of isolation, gentamicin, cephalexin, cefotaxime, erythromycin, clindamycin, lincomycin, tobramycin, ceftriaxone and streptomycin in 100%. Antibiotic resistance correlates with the clinical failure of antimicrobials in veterinary medicine. Multi-resistance of Enterococcus faecalis to various groups of antibacterial agents leads to limited use of antibiotics, which significantly reduces the possibilities of therapy and affects the economic condition of agriculture.

A Two-Step Single Plex PCR Method for Evaluating Key Colonic Microbiota Markers in Young Mexicans with Autism Spectrum Disorders: Protocol and Pilot Epidemiological Application.

Many neurological disorders have a distinctive colonic microbiome (CM) signature. Particularly, children with autism spectrum disorders (ASD) exhibit a very dissimilar CM when compared to neurotypical (NT) ones, mostly at the species level. Thus far, knowledge on this matter comes from high-throughput (yet very expensive and time-consuming) analytical platforms, such as massive high-throughput sequencing of bacterial 16S rRNA. Here, pure (260/280 nm, ~1.85) stool DNA samples (200 ng.µL-1) from 48 participants [39 ASD, 9 NT; 3-13 y] were used to amplify four candidate differential CM markers [Bacteroides fragilis (BF), Faecalibacterium prausnitzii (FP), Desulfovibrio vulgaris (DV), Akkermansia muciniphila (AM)], using micro-organism-specific oligonucleotide primers [265 bp (BF), 198 bp (FP), 196 bp (DV), 327 bp (AM)] and a standardized two-step [low (step 1: °Tm-5 °C) to high (stage 2: °Tm-0 °C) astringent annealing] PCR protocol (2S-PCR). The method was sensitive enough to differentiate all CM biomarkers in the studied stool donors [↑ abundance: NT (BF, FP, AM), ASD (DV)], and phylogenetic analysis confirmed the primers' specificity.

Echinacoside inhibits colorectal cancer metastasis via modulating the gut microbiota and suppressing the PI3K/AKT signaling pathway

Ethnopharmacological relevanceEchinacoside (ECH) is the dominant phenylethanoid glycoside-structured compound identified from our developed herbal formula Huangci granule, which has been previously reported to inhibit the invasion and metastasis of CRC and prolong patients’ disease-free survival duration. Though ECH has inhibitory activity against aggressive colorectal cancer (CRC) cells, its anti-metastasis effect in vivo and the action mechanism is undetermined. Given that ECH has an extremely low bioavailability and gut microbiota drives the CRC progression, we hypothesized that ECH could inhibit metastatic CRC by targeting the gut microbiome. Aim of the studyThe purpose of this study was to investigate the impact of ECH on colorectal cancer liver metastasis in vivo and its potential mechanisms. Materials and methodsAn intrasplenic injection-induced liver metastatic model was established to examine the efficiency of ECH on tumor metastasis in vivo. Fecal microbiota from the model group and the ECH group were separately transplanted into pseudo-sterile CRLM mice in order to verify the role of gut flora in the ECH anti-metastatic effect. The 16S rRNA gene sequence was applied to analyze the structure and composition of the gut microbiota after ECH intervention, and the effect of ECH on short-chain fatty acid (SCFAs)-producing bacteria growth was proven by anaerobic culturing in vitro. GC-MS was applied to quantitatively analyze the serum SCFAs levels in mice. RNA-seq was performed to detect the gene changes involving tumor-promoting signaling pathway. ResultsECH inhibited CRC metastasis in a dose-dependent manner in the metastatic colorectal cancer (mCRC) mouse model. Manipulation of gut bacteria in the mCRC mouse model further proved that SCFA-generating gut bacteria played an indispensable role in mediating the antimetastatic action of ECH. Under an anaerobic condition, ECH benefited SCFA-producing microbiota without affecting the total bacterial load, presenting a dose-dependent promotion on the growth of a butyrate producer, Faecalibacterium prausnitzii (F.p). Furthermore, ECH-reshaped or F.p-colonized microbiota with a high butyrate-producing capability inhibited liver metastasis by suppressing PI3K/AKT signaling and reversing the epithelial-mesenchymal transition (EMT) process, whereas this anti-metastatic ability was abrogated by the butyrate synthase inhibitor heptanoyl-CoA. ConclusionThis study demonstrated that ECH exhibits oral anti-metastatic efficacy by facilitating butyrate-producing gut bacteria, which downregulates PI3K/AKT signaling and EMT. It hints at a novel role for ECH in CRC therapy.

Age-related effects on the modulation of gut microbiota by pectins and their derivatives: an in vitro study.

The present study investigates whether supplementation with pectin-type polysaccharides has potential to improve aging-associated dysbiosis of the gut microbiota. The influence of different types of pectins on the gut microbiota composition and short-chain fatty acids (SCFAs) profiles of elderly was compared to younger adults. Pectins studied included a pectin polysaccharide (PEC), a partially hydrolyzed pectin (PPH), and a pectin oligosaccharide (POS). Additionally, inulin was used as a reference prebiotic substrate. Individual fecal samples were collected from healthy elderly volunteers (70-75 years) and younger adults (30-35 years). In vitro fermentations were performed using the CoMiniGut model with controlled temperature and pH. Samples were withdrawn at baseline and after 24 h fermentation for measurement of SCFAs production and microbiota composition by 16S rRNA gene sequencing. The results showed that fermentations with PEC and PPH resulted in a specific stimulation of Faecalibacterium prausnitzii regardless of the age groups. Collinsella aerofaciens became a dominating species in the young adult group with fermentations of all three pectins, which was not observed in the elderly group. No significant differences in SCFAs production were found among the pectins, indicating a high level of functional redundancy. Pectins boosted various bacterial groups differently from the reference prebiotic substrate (inulin). We also found inulin had reduced butyrogenic and bifidogenic effects in the elderly group compared to the younger adult group. In conclusion, the in vitro modulating effects of pectins on elderly gut microbiota showed potential of using pectins to improve age-related dysbiosis.

The profile of key gut microbiota members and short-chain fatty acids in patients with sepsis

Sepsis is a complex clinical disorder with heterogeneous etiological factors. Given its high mortality rate, it is considered a global health issue. Recently, the link between gut microbiota and their metabolites, especially short-chain fatty acids, in the pathophysiology of sepsis has been reported. However, there are few findings to confirm this relationship. This study aimed to evaluate some key gut microbiota members, pathogenic bacteria, and short-chain fatty acids in non-ICU patients with sepsis caused by bacteremia compared to a control group. In this case-control study, 45 stool samples from patients with sepsis and 15 healthy persons were collected from October 2021 to August 2022 in Tabriz, Iran. The position of some gut microbiota members and the main short-chain fatty acids concentration were assessed in the two groups by the Q-PCR and the high-performance liquid chromatography system. Faecalibacterium prausnitzii and Bifidobacterium sp. As bacterial with protective features in non-ICU patients with sepsis decreased significantly. Moreover, the concentrations of acetic acid and propionic acid significantly decreased in this group compared to the healthy volunteers. In contrast, the pathogenic bacteria members such as Enterobacteriaceae and Bacteroides sp. Increased significantly in the patients compared to the healthy individuals. The concentration of butyric acid decreased in the patients, but this change was not significant in the two groups. Protective and immune functions of F. prausnitzii and Bifidobacterium sp., as well as acetate and propionate, are evident. In this investigation, this profile was significantly reduced in non-ICU patients with sepsis compared to the control group.

Milk and dairy product intakes, intestinal bacteria, and respiratory infections in children of elementary school age and older in Japan

ObjectivesThe aim of this study was to examine the associations between milk and dairy product intakes, intestinal bacteria, and respiratory infections in children of elementary school age and older in Japan. MethodsWe conducted cross-sectional surveys each year from 2013 to 2015 for grades 2, 5, and 8 students of an elementary and junior high school (n = 1020). Exclusion owing to ineligibility regarding data on dietary intake, respiratory infections, and intestinal bacteria led to 922 participants for the analyses. Dietary intake was assessed with a self-administered food frequency questionnaire. Respiratory infections occurring ≥ 4 episodes over the past year were determined based on the caregivers’ reports. Intestinal bacteria (species and counts) were analyzed with real-time polymerase chain reaction. Logistic regression models were used to estimate the odds ratios (ORs) and 95% CIs. ResultsThe odds of ≥ 4 respiratory infection episodes decreased with higher milk intake after adjusting for potential confounders, and the ORs (95% CIs) for the second and third tertile categories, compared with the first tertile category, were 0.91 (0.58–1.42) and 0.48 (0.29–0.77), respectively (P for trend = 0.001). A decreasing trend in the ORs for lactic acid drink intake was observed only in those with a low count of intestinal Faecalibacterium prausnitzii. ConclusionsWe found that higher milk intake was inversely associated with respiratory infections in children older than preschool age. Higher lactic acid drink intake could be inversely associated only in children with a low F. prausnitzii count in the intestine.

Pea hull fiber supplementation does not modulate uremic metabolites in adults receiving hemodialysis: a randomized, double-blind, controlled trial.

Fiber is a potential therapeutic to suppress microbiota-generated uremic molecules. This study aimed to determine if fiber supplementation decreased serum levels of uremic molecules through the modulation of gut microbiota in adults undergoing hemodialysis. A randomized, double-blinded, controlled crossover study was conducted. Following a 1-week baseline, participants consumed muffins with added pea hull fiber (PHF) (15 g/d) and control muffins daily, each for 4 weeks, separated by a 4-week washout. Blood and stool samples were collected per period. Serum p-cresyl sulfate (PCS), indoxyl sulfate (IS), phenylacetylglutamine (PAG), and trimethylamine N-oxide (TMAO) were quantified by LC-MS/MS, and fecal microbiota profiled by 16S rRNA gene amplicon sequencing and specific taxa of interest by qPCR. QIIME 2 sample-classifier was used to discover unique microbiota profiles due to the consumption of PHF. Intake of PHF contributed an additional 9 g/d of dietary fiber to the subjects' diet due to compliance. No significant changes from baseline were observed in serum PCS, IS, PAG, or TMAO, or for the relative quantification of Akkermansia muciniphila, Faecalibacterium prausnitzii, Bifidobacterium, or Roseburia, taxa considered health-enhancing. Dietary protein intake and IS (r = -0.5, p = 0.05) and slow transit stool form and PCS (r = 0.7, p < 0.01) were significantly correlated at baseline. PHF and control periods were not differentiated; however, using machine learning, taxa most distinguishing the microbiota composition during the PHF periods compared to usual diet alone were enriched Gemmiger, Collinsella, and depleted Lactobacillus, Ruminococcus, Coprococcus, and Mogibacteriaceae. PHF supplementation did not mitigate serum levels of targeted microbial-generated uremic molecules. Given the high cellulose content, which may be resistant to fermentation, PHF may not exert sufficient effects on microbiota composition to modulate its activity at the dose consumed.

Effect of Plasma-Activated Water and Direct Plasma on Enterococcus faecalis Bacteria for Disinfection of Tooth Root Canal

This research studies the effect regarding two plasma types, plasma jet and plasma-activated water (PAW), on tooth root canal bacteria Enterococcus faecalis. The plasma jet works with argon gas, and it is generated by a power supply that operates at alternating high voltages in the form of a sinusoidal wave with peak-to-peak value of about 12 kV at a frequency of 30 KHz and its power is about 200 watts. This plasma was utilized directly to treat the tooth canal and indirectly by activating the water that was used later for treating the Enterococcus faecalis bacteria that are present in the tooth root. Pure distilled water was treated by plasma jet for one hour at flow rate 1 . Plasma water activated by plasma contains NO2, NO3, and H2O2 with a concentration (10,100,200) ppm, respectively. It was noted, as a result of direct exposure with the two values of gas flow rate and for different exposure times, an ideal killing rate (from 298 x 108 to zero)CFU/ml was obtained when the exposure time changed from (5-30) minutes with a constant gas flow rate of 2.5 ; while in the case of changing the gas flow rate from (0.5-2.5) and fixing time to 5 min, the killing rate decreased from (298 ×108 to 30×108) CFU/ml. In the case of (PAW) when the time changed (5-25) min, an ideal killing rate was obtained (298 x 108 to zero) CFU/ml; adopted one factor, where the results indicated that plasma-activated water is good in disinfecting the root canal of the tooth and inactivating bacteria.

Maternal Magnolol Supplementation during Pregnancy and Lactation Promotes Antioxidant Capacity, Improves Gut Health, and Alters Gut Microbiota and Metabolites of Weanling Piglets.

Maternal nutrition exerts a profound effect on the postnatal performance of offspring, especially during the weaning period. The multifunctional bioactive component magnolol (MAG) has shown promise as a dietary supplement. This study aimed to explore the effects of maternal MAG supplementation on the antioxidant capacity, gut health, gut microbiome, and metabolome composition of weanling piglets. Fifty pregnant sows were randomly divided into two equally sized groups, the control group and the group supplemented with 100 g/t MAG during the gestation and lactation periods, and 7 days postweaning, the pups were euthanized. The microbiome and metabolome features of weanling piglet colons were compared. Our results revealed that maternal MAG supplementation modified the serum redox status of weanling piglets by decreasing malondialdehyde concentration and increasing superoxide dismutase activity and total antioxidant capacity. Moreover, the decreased indicators of diarrhea were accompanied by improved gut barrier function, in which serum diamine oxidase concentration was decreased, and expressions of zona occludens-1, claudin-1, and intestinal alkaline phosphatase were increased in the colon of weanling piglets from sows supplemented with MAG. Further analysis of the gut microbiota indicated that maternal MAG supplementation significantly increased the relative abundance of beneficial bacteria in the colon of weanling piglets, including Faecalibacterium prausnitzii and Oscillospira. Metabolome analysis identified 540 differential metabolites in the colon of piglets from MAG-fed dams, of which glycerophospholipid classes were highly correlated with progeny gut health and key beneficial bacteria. Our findings indicated that maternal MAG supplementation can improve the oxidative status and gut health of weanling piglets, possibly due to alterations in the gut microbiota and metabolites.

Multistrain Probiotics with Fructooligosaccharides Improve Middle Cerebral Artery Occlusion-Driven Neurological Deficits by Revamping Microbiota-Gut-Brain Axis.

Recent burgeoning literature unveils the importance of gut microbiota in the neuropathology of post-stroke brain injury and recovery. Indeed, ingestion of prebiotics/probiotics imparts positive effects on post-stroke brain injury, neuroinflammation, gut dysbiosis, and intestinal integrity. However, information on the disease-specific preference of selective prebiotics/probiotics/synbiotics and their underlying mechanism is yet elusive. Herein, we examined the effect of a new synbiotic formulation containing multistrain probiotics (Lactobacillus reuteri UBLRu-87, Lactobacillus plantarum UBLP-40, Lactobacillus rhamnosus UBLR-58, Lactobacillus salivarius UBLS-22, and Bifidobacterium breve UBBr-01), and prebiotic fructooligosaccharides using a middle cerebral artery occlusion (MCAO) model of cerebral ischemia in female and male rats. Three weeks pre-MCAO administration of synbiotic rescinded the MCAO-induced sensorimotor and motor deficits on day 3 post-stroke in rotarod, foot-fault, adhesive removal, and paw whisker test. We also observed a decrease in infarct volume and neuronal death in the ipsilateral hemisphere of synbiotic-treated MCAO rats. The synbiotic treatment also reversed the elevated levels/mRNA expression of the glial fibrillary acidic protein (GFAP), NeuN, IL-1β, TNF-α, IL-6, matrix metalloproteinase-9, and caspase-3 and decreased levels of occludin and zonula occludens-1 in MCAO rats. 16S rRNA gene-sequencing data of intestinal contents indicated an increase in genus/species of Prevotella (Prevotella copri), Lactobacillus (Lactobacillus reuteri), Roseburia, Allobaculum, and Faecalibacterium prausnitzii, and decreased abundance of Helicobacter, Desulfovibrio, and Akkermansia (Akkermansia muciniphila) in synbiotic-treated rats compared to the MCAO surgery group. These findings confer the potential benefits of our novel synbiotic preparation for MCAO-induced neurological dysfunctions by reshaping the gut-brain-axis mediators in rats.

Effects of the Phytocompound Combination against Dysbiosis Induced by AGE-Rich High-fat Diet in Mice

Background and Aims: The composition of the Microbiota can be influenced by various lifestyle and environmental factors, including diet. We designed a study to investigate the improving effects of plant extract, the combination of turmeric, ginger, boswellia, and cat’s claw on the abundance of key members of the gut microbiota, Bacteroidetes, Akkermansia muciniphila (A. muciniphila), Faecalibacterium prausnitzii (F. prausnitzii), Firmicutes and Bifidobacterium in mice treated with advanced glycation end products (AGE) rich high-fat diet (HFD).&#x0D; Materials and Methods: Eighteen 2-month age male C57BL/6 mice were adopted to a regular diet for 1 week and then fed with an HFD or regular diet. After 8 weeks of diet, animals received plant extract concurrently with HFD for 8 weeks. Stools were taken, DNA of stool samples was extracted, and qPCR of 16s rDNA universal primers was performed. Then the effect of plant extract on dysbiosis induced by AGE-rich HFD was accessed.&#x0D; Results: Our results revealed the frequencies of Bacteroidetes (p=0.001), A. muciniphila (p= 0.0005), F. prausnitzii (p&lt;0.0001), and Bifidobacterium (p= 0.0008) were reduced, whereas the frequency of Firmicute was increased in the AGE-rich HFD group. A significant increase in the F/B ratio was observed in the HFD group than in the regular diet group (p= 0.0003). plant extract reduced the F/B ratio and improved gut microbiota homeostasis. &#x0D; Conclusion: Plant extract restored gut microbiota patterns in HFD-treated mice. It seems more studies are required to prove the application of plant extracts for modulating the gut microbiota as a promising new biomarker with potential for therapeutic applications.

Antibacterial and smear layer removal efficacy of moringa (Moringa oleifera): An in vitro study

ObjectiveThis study evaluated the effectiveness of moringa (Moringa oleifera) leaves decoction for removing a smear layer compared to sodium hypochlorite (NaOCl) and ethylenediaminetetraacetic acid (EDTA), as well as its antimicrobial activities. MethodsThe moringa leaves were extracted using hot water decoction at two different concentrations (2.5% and 5.0% w/v). A total of 30 extracted human single-rooted teeth were prepared to assess the smear layer removal efficacy. The presence of a smear layer in the middle third of the root canal was detected by confocal microscopy. Then the antibacterial effects were assessed against Enterococcus faecalis and Streptococcus mutans bacteria using the agar diffusion method. ResultsThe 2.5% and 5.0% decoction were significantly more effective than 0.25% NaOCl in removing the smear layer (p < 0.05); however, no significant difference was observed compared to EDTA (p > 0.05). The in vitro antimicrobial assay showed that 5.0% decoction had higher antimicrobial activity against both of the test pathogens. ConclusionThe findings of this study suggest that moringa leaves decoction can be considered an effective irrigant in endodontics.

H2 generated by fermentation in the human gut microbiome influences metabolism and competitive fitness of gut butyrate producers

BackgroundHydrogen gas (H2) is a common product of carbohydrate fermentation in the human gut microbiome and its accumulation can modulate fermentation. Concentrations of colonic H2 vary between individuals, raising the possibility that H2 concentration may be an important factor differentiating individual microbiomes and their metabolites. Butyrate-producing bacteria (butyrogens) in the human gut usually produce some combination of butyrate, lactate, formate, acetate, and H2 in branched fermentation pathways to manage reducing power generated during the oxidation of glucose to acetate and carbon dioxide. We predicted that a high concentration of intestinal H2 would favor the production of butyrate, lactate, and formate by the butyrogens at the expense of acetate, H2, and CO2. Regulation of butyrate production in the human gut is of particular interest due to its role as a mediator of colonic health through anti-inflammatory and anti-carcinogenic properties.ResultsFor butyrogens that contained a hydrogenase, growth under a high H2 atmosphere or in the presence of the hydrogenase inhibitor CO stimulated production of organic fermentation products that accommodate reducing power generated during glycolysis, specifically butyrate, lactate, and formate. Also as expected, production of fermentation products in cultures of Faecalibacterium prausnitzii strain A2-165, which does not contain a hydrogenase, was unaffected by H2 or CO. In a synthetic gut microbial community, addition of the H2-consuming human gut methanogen Methanobrevibacter smithii decreased butyrate production alongside H2 concentration. Consistent with this observation, M. smithii metabolic activity in a large human cohort was associated with decreased fecal butyrate, but only during consumption of a resistant starch dietary supplement, suggesting the effect may be most prominent when H2 production in the gut is especially high. Addition of M. smithii to the synthetic communities also facilitated the growth of E. rectale, resulting in decreased relative competitive fitness of F. prausnitzii.ConclusionsH2 is a regulator of fermentation in the human gut microbiome. In particular, high H2 concentration stimulates production of the anti-inflammatory metabolite butyrate. By consuming H2, gut methanogenesis can decrease butyrate production. These shifts in butyrate production may also impact the competitive fitness of butyrate producers in the gut microbiome.3hTM6nso89x7h1g_xY7TKkVideo

#3953 TOXIC MICROBIOME AND CHRONIC KIDNEY DISEASE: INSIGHTS FROM THE CKD-REIN COHORT STUDY

Abstract Background and Aims Many uremic toxins (UTs) originate from gut microbiome, and contribute to chronic kidney disease (CKD) progression and cardiovascular morbidity. In order to reduce uremic symptoms and CKD progression, patients have several dietary restrictions, which may influence gut microbiome composition, and impact UTs production. An altered microbiome may contribute to UTs increase in those patients. However, the role of key bacterial taxa in producing UTs and the impact of diet on UTs variance in non-dialyzed patients are not well known. The objectives of this study were, first, to compare microbial features between CKD patients and healthy controls, and, second, to investigate the relation of gut microbiome with uremic toxicity, as well as the potential impact of diet on such relationship. Method Characterization of gut metagenomes, 10 UTs and 3 precursors’ serum concentrations by LC-MS/MS, host characteristics and diet were obtained from 240 non-dialysis CKD patients from the CKD-REIN cohort (mean ± SD): age: 68 ± 11 years, 71% male, estimated glomerular filtration rate (eGFR): 33.2 ± 12.7 ml/min/1.73m². First, to identify microbial biomarkers characterizing the gut microbiome-related toxicity in CKD, we compared microbiome features between 78 CKD patients and 78 age-, sex-, and BMI-matched healthy controls from the Milieu Interieur (MI) cohort: age: 58 ± 10 years, 60% male, eGFR: 89 ± 13. Second, we performed a multiomics’ data integration analysis via a supervised modelling to investigate cross-sectionally the association between host characteristics, gut microbiome, UTs, and diet-related features according to CKD severity (eGFR&amp;lt;30, n = 110 vs eGFR ≥30 mL/min/1.73m², n = 130). Results Compared to healthy controls, CKD patients had a significant reduced gut microbiome health index. Several Metagenomic Species Pan-genomes (MSPs) were significantly contrasted between MI and CKD cohorts: 43 species were enriched in CKD patients vs 24 in controls. Species most enriched in CKD patients included several UTs producers such as Lachnospiraceae spp, Dysosmobacter – Oscillibacter spp, Butyricimonas faecihominis, Victivallis vadensis and Hungatella spp, some of which were positively correlated with the following UTs: 3-Carboxy-4-methyl-5-propyl-2-furanpropionate (CMPF), trimethylamine-N-oxide (TMAO), and indole-3-acetic acid (3-IAA). Moreover, species belonging to Enterocloster and Hungatella genera (both members of Lachnospiraceae family) were found to be negatively correlated with eGFR. Among species associated with CKD severity, species carrying genes for UTs production were observed such as Desuflovibiro fairfieldensis, Bacteroides clarus and Blautia obeum along with increasing alcohol and hot drinks consumption, CRP and several UTs (kynurenic acid, indoxyl sulfate and Phenylacetylglutamine) levels. In contrast, some taxa like Faecalibacterium prausnitzii and Dysosmobacter welbionis were associated with legume intake but not with UTs. Conclusion Our study highlights an alteration of gut microbiome in CKD patients compared to healthy controls, with increased abundance of UTs producer species. The results of the multidimensional data integration modelling suggest a strong interplay between food intake, gut microbiome modifications, UTs accumulation and clinical features. These findings might open to promising therapeutic strategies to reduce microbiome-related toxicity.