Dysbiotic Flora Research Articles

The Impact of the Microbiological Vaginal Swab on the Reproductive Outcome in Infertile Women.

The thesis on which this paper is based intended to investigate whether the result of the microbiological vaginal swab has an influence on the outcome of the fertility treatment. The microbiological vaginal swabs of patients who received fertility treatment at Saarland University Hospital were evaluated. Depending on the microorganisms detected, the swab result was classified as inconspicuous, intermediate, or conspicuous. The SPSS software was used to determine the correlation between the swab result and the outcome of the fertility treatment. Dysbiosis was associated with a worse outcome of fertility treatment. The pregnancy rate with a conspicuous swab was 8.6%, whereas it was 13.4% with an inconspicuous swab. However, this association was not statistically significant. Furthermore, an association of endometriosis with dysbiosis was found. Endometriosis was more frequent with a conspicuous swab result than with an inconspicuous result (21.1% vs. 17.7%), yet the correlation was not statistically significant. However, the absence of lactobacilli was significantly associated with endometriosis (p = 0.021). The association between endometriosis and a lower pregnancy rate was also statistically significant (p = 0.006). The microbiological vaginal and cervical swabs can be used as predictors for the success of fertility treatments. Further studies are needed to assess the impact of transforming a dysbiotic flora into a eubiotic environment on the success of fertility treatments.

P: 12 PROFIT: PROspective, Randomised Placebo-controlled Feasibility Trial of Faecal Microbiota Transplantation in Cirrhosis Interim Analysis

BACKGROUND: Patients with cirrhosis have intestinal dysbiosis and small bowel bacterial overgrowth, with enhanced gut permeability, allowing translocation of pathogen-associated molecules and viable bacteria into the systemic circulation. Faecal Microbiota Transplantation (FMT) seeks to replace the dysbiotic flora with healthy microbes, to reduce systemic inflammation and improve transplant-free outcomes. PROFIT is a single-blinded randomised placebo-controlled trial of FMT in 32 patients with advanced stable cirrhosis. The primary outcome of the study was to assess the safety and feasibility of a single naso-jejunally administered FMT or placebo (3:1 allocation) in patients with a MELD score between 10 and 16. METHODS: To date eighteen patients have been treated (FMT n = 13; placebo n = 5). 71% of those treated were male [mean age 54.4 years (range 38–75) and mean MELD 12.88 (range 10–16)]. Patients continued lactulose, but were required to discontinue all antibiotics, including rifaximin, two weeks prior to treatment. Following bowel preparation with Moviprep®, 200 mL FMT/placebo was delivered via naso-jejunal tube in to the proximal jejunum at gastroscopy. Patients were assessed for adverse events and underwent clinical review at day 7, 30 and 90 with biobanking of stool, saliva, urine, whole blood and plasma for quantitative metagenomics, metabonomics, bacterial DNA quantification, bile acids, whole blood culture in response to endotoxin and plasma cytokines and leucocyte function testing at a later date. RESULTS: The FMT has been well tolerated and the five serious adverse events (SAEs) reported requiring hospital admission (placebo n = 1; FMT n = 4) were not treatment-related. Biochemistry, leucocyte count and MELD score did not change. Mean venous ammonia reduced at day 30 in the FMT cohort [71.67 μmol/L (95% CI 54.43–88.9) day 0 versus 51.22 μmol/L (CI 35.18–67.26) day 30] whereas it increased in the placebo group [54.4 μmol/L (95% CI 38.17–70.63) day 0 versus 73.25 μmol/L (95% CI 22.54–124) at day 30]. Figure 1 shows a reduction in delta ammonia at day 7 [mean −1.15 (−16.23 to 13.73 95% CI)] versus day 30 [−15.11 (−2.82 to −27.4) post FMT. CONCLUSIONS: The interim analysis shows that nasojejunally administered FMT appears safe and well tolerated in patients with advanced cirrhosis and reduces venous ammonia at day 30.

Survival of an Aggregatibacter actinomycetemcomitans quorum sensing luxS mutant in the mouths of Rhesus monkeys: insights into ecological adaptation.

Experiments were designed to explore a prominent autoinducer-2 (AI-2) producing gene (luxS) related to colonization and survival of Aggregatibacter actinomycetemcomitans, a low abundance member of the indigenous flora, that forms a key component of the dysbiotic flora in localized aggressive periodontitis. The luxS gene was disrupted in a primate strain of A.actinomycetemcomitans before implantation into the oral cavity of Rhesus monkeys (Rh). The colonization efficiency of the luxS mutant (RhAa-VS4) was compared with the parental wild-type strain (RhAa3) (positive control) and a ltxA mutant (RhAa-VS2) (negative control). The in vivo results showed that the luxS mutation had minimal impact on A.actinomycetemcomitans colonization compared with the wild-type RhAa3 strain. In vitro studies revealed that there was a significant upregulation of attachment-related genes aae, apiA, and flp in the RhAa-VS4 strain compared with RhAa3. Biofilm forming ability was also significantly increased in the RhAa-VS4 strain compared with RhAa3, whereas the AI-2 signal was ablated. The exogenous addition of the AI-2 precursor dihydroxy pentanedione allowed the RhAa-VS4 strain to achieve RhAa3 biofilm levels. This is the first primate study to test the relevance of LuxS in vivo. In vitro assessment suggests that in vivo survival of the RhAa-VS4 strain was due to the production of signaling AI-2 molecules derived from other members of the flora as well as the upregulation of genes related to attachment and biofilm formation.

Dysbiosis and Staphylococcus aureus Colonization Drives Inflammation in Atopic Dermatitis

Staphylococcus aureus skin colonization is universal in atopic dermatitis and common in cancer patients treated with epidermal growth factor receptor inhibitors. However, the causal relationship of dysbiosis and eczema has yet to be clarified. Herein, we demonstrate that Adam17(fl/fl)Sox9-(Cre) mice, generated to model ADAM17-deficiency in human, developed eczematous dermatitis with naturally occurring dysbiosis, similar to that observed in atopic dermatitis. Corynebacterium mastitidis, S. aureus, and Corynebacterium bovis sequentially emerged during the onset of eczematous dermatitis, and antibiotics specific for these bacterial species almost completely reversed dysbiosis and eliminated skin inflammation. Whereas S. aureus prominently drove eczema formation, C. bovis induced robust T helper 2 cell responses. Langerhans cells were required for eliciting immune responses against S. aureus inoculation. These results characterize differential contributions of dysbiotic flora during eczema formation, and highlight the microbiota-host immunity axis as a possible target for future therapeutics in eczematous dermatitis.

Enteric Commensal Bacteria Induce Extracellular Signal-regulated Kinase Pathway Signaling via Formyl Peptide Receptor-dependent Redox Modulation of Dual Specific Phosphatase 3

The normal microbial occupants of the mammalian intestine are crucial for maintaining gut homeostasis, yet the mechanisms by which intestinal cells perceive and respond to the microbiota are largely unknown. Intestinal epithelial contact with commensal bacteria and/or their products has been shown to activate noninflammatory signaling pathways, such as extracellular signal-related kinase (ERK), thus influencing homeostatic processes. We previously demonstrated that commensal bacteria stimulate ERK pathway activity via interaction with formyl peptide receptors (FPRs). In the current study, we expand on these findings and show that commensal bacteria initiate ERK signaling through rapid FPR-dependent reactive oxygen species (ROS) generation and subsequent modulation of MAP kinase phosphatase redox status. ROS generation induced by the commensal bacteria Lactobacillus rhamnosus GG and the FPR peptide ligand, N-formyl-Met-Leu-Phe, was abolished in the presence of selective inhibitors for G protein-coupled signaling and FPR ligand interaction. In addition, pretreatment of cells with inhibitors of ROS generation attenuated commensal bacteria-induced ERK signaling, indicating that ROS generation is required for ERK pathway activation. Bacterial colonization also led to oxidative inactivation of the redox-sensitive and ERK-specific phosphatase, DUSP3/VHR, and consequent stimulation of ERK pathway signaling. Together, these data demonstrate that commensal bacteria and their products activate ROS signaling in an FPR-dependent manner and define a mechanism by which cellular ROS influences the ERK pathway through a redox-sensitive regulatory circuit.

The Microbatome and Intestinal Microflora in Diverticular Disease

The microflora of the human gastrointestinal tract contain 10 14 organisms, more bacteria than make up the body. The largest number is in the colon, and the anaerobes outnumber the aerobes by 100 or 1000:1. The anaerobic species of Bacteroides, Eubacterium, and Bifidobacterium usually predominate, and usually the most common organism is Faecalibacterium prausnitzii. Other details are described in this study. Of note is that there are changes in the normal flora in patients suffering from inflammatory bowel disease and the irritable bowel syndrome. As diverticular disease and diverticulitis seem to have some chronic inflammatory component to their pathophysiology, it is suspicious that there is a dysbiosis, but we have not had studies published on the flora in diverticular disease. Further suspicion of the role of a dysbiotic flora is raised since the initial reports that both mesalamine, an anti-inflammatory agent, and probiotics, either Escherichia coli Nissle or Lactobacillus casei, have been effective in controlling recurrent attacks. The details of these studies are described.

Treating Irritable Bowel Syndrome with a Food Elimination Diet Followed by Food Challenge and Probiotics

Objective: In Irritable Bowel Syndrome, the gut-associated immune system may be up-regulated resulting in immune complex production, low-grade inflammation, loss of Class I bacteria, and translocation of inflammatory mediators and macromolecules outside of the GI lumen. Since food intolerance may be one of the reasons for this upregulation, our goal was to investigate the role of food intolerance in IBS patients.Methods: In this open label pilot study, we enrolled 20 patients with IBS by Rome II criteria (15 women, ages 24–81) who had failed standard medical therapies in a tertiary care GI clinic. Baseline serum IgE and IgG food and mold panels, and comprehensive stool analysis (CSA) were performed. Breath-hydrogen testing and IBS Quality-of-Life (QOL) questionnaires were obtained. Patients underwent food elimination diets based on the results of food and mold panels followed by controlled food challenge. Probiotics were also introduced. Repeat testing was performed at 6-months. We followed up with this cohort at 1 year after trial completion to assess the reported intervention and for placebo effect.Results: Baseline abnormalities were identified on serum IgG food and mold panels in 100% of the study subjects with significant improvement after food elimination and rotation diet (p < 0.05). Significant improvements were seen in stool frequency (p < 0.05), pain (p < 0.05), and IBS-QOL scores (p < 0.0001). Imbalances of beneficial flora and dysbiotic flora were identified in 100% of subjects by CSA. There was a trend to improvement of beneficial flora after treatment but no change in dysbiotic flora. The 1-year follow up demonstrated significant continued adherence to the food rotation diet (4.00 ± 1.45), minimal symptomatic problems with IBS (4.00 ± 1.17), and perception of control over IBS (4.15 ± 1.23). The continued use of probiotics was considered less helpful (3.40 ± 1.60).Conclusion: These data demonstrate that identifying and appropriately addressing food sensitivity in IBS patients not previously responding to standard therapy results in a sustained clinical response and impacts on overall well being and quality of life in this challenging entity.