Overcoming donor variability and risks associated with fecal microbiota transplants through bacteriophage-mediated treatments

bioRxiv (Cold Spring Harbor Laboratory), Journal Year: 2023, Volume and Issue: unknown

Published: March 17, 2023

ABSTRACT Background Fecal microbiota transplantation (FMT) and fecal virome (FVT, sterile filtrated donor feces) have been effective in treating recurrent Clostridioides difficile infections, possibly through bacteriophage-mediated modulation of the gut microbiome. However, challenges like variability, costly screening, coupled with concerns over pathogen transfer (incl. eukaryotic viruses) FMT or FVT hinders their wider clinical application less acute diseases. Methods To overcome these challenges, we developed methods to broaden FVT’s while maintaining efficacy increasing safety. Specifically, employed following approaches: 1) Chemostat-fermentation reproduce bacteriophage component remove viruses (FVT-ChP), 2) solvent-detergent treatment inactivate enveloped (FVT-SDT), 3) pyronin-Y inhibit RNA-virus replication (FVT-PyT). We assessed processed FVTs a C. infection mouse model compared them untreated (FVT-UnT), FMT, saline. Results FVT-SDT, FVT-UnT, FVT-ChP reduced incidence mice reaching humane endpoint (0/8, 2/7, 3/8, respectively) FVT-PyT, saline control (5/8, 7/8, 5/7, significantly load colonizing cells toxin A/B levels. There was potential elimination colonization, 7 out 8 treated FVT-SDT testing negative qPCR. In contrast, all other treatments exhibited continued presence . Moreover, results were supported by changes microbiome profiles, cecal cytokine levels histopathological findings. Assessment viral engraftment FMT/FVT host-phage correlations analysis suggested that phages likely an important contributing factor associated efficacy. Conclusions This proof-of-concept study show specific modifications hold promise addressing related variability risks. Two strategies lead limiting colonization mice, solvent/detergent chemostat-propagation emerging as promising approaches.

Language: Английский

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Evaluating Bacterial Viability in Faecal Microbiota Transplantation: A Comparative Analysis of In Vitro Cultivation and Membrane Integrity Methods

bioRxiv (Cold Spring Harbor Laboratory), Journal Year: 2024, Volume and Issue: unknown

Published: Feb. 9, 2024

Abstract Background Faecal microbiota transplantation (FMT) is a developing therapy for disorders related to gut dysbiosis. Despite its growing application, standardized protocols FMT filtrate preparation and quality assessment remain undeveloped. The viability of bacteria in the crucial FMT’s efficacy validating protocol execution. We compared two methods—in vitro cultivation membrane integrity assessment—for their accuracy, reproducibility, clinical applicability measuring bacterial frozen stool filtrate. Methods Bacterial was evaluated using (i) through fluorescent DNA staining with SYTO9 propidium iodide, followed by flow cytometry; (ii) culturable counts (colony-forming units, CFU) under aerobic or anaerobic conditions. Results refined integrated cytometry samples. Both integrity-based cultivation-based methods exhibited significant variability across different filtrates, without correlation. method showed mean coefficient variance 17%, ranging from 5.3% 52.9%. Conversely, approach yielded highly reproducible results, median viable cells 0.9%, 8.5% 0.04%. Conclusion cultivation-dependent produces inconsistent outcomes. In contrast, offers robust precise data, making it option routine faecal material evaluation FMT.

Language: Английский

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Multi-Omics Analysis Unravels the Impact of Stool Sample Logistics on Metabolites and Microbial Composition

Microorganisms, Journal Year: 2024, Volume and Issue: 12(10), P. 1998 - 1998

Published: Sept. 30, 2024

Human health and the human microbiome are inevitably intertwined, increasing their relevance in clinical research. However, collection, transportation storage of faecal samples may introduce bias due to methodological differences, especially since postal shipping is a common practise large-scale cohort studies. Using four different Omics layer, we determined structural (16S rRNA sequencing, cytometric microbiota profiling) functional integrity (SCFAs, global metabolome) relation easy-to-handle conditions. These conditions were at -20 °C, °C as glycerol stock, 4 room temperature with without oxygen exposure for maximum one week. Storage time affected on all levels. magnitude was donor-dependent, highlighting need purpose-optimized sample collection multi-donor The effects negligible analyses. At ambient temperature, SCFA compositional profiles stable 24 h 48 h, respectively, while maintained h. metabolome highly susceptible, already changing non-frozen Thus, best preserved levels when transported native frozen within leading least biased outcomes analysis. We conclude that immediate freezing stool lab suited planned multi-Omics analyses include metabolomics extend standard sequencing approaches.

Language: Английский

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Fecal Microbiota Transplantation: A Systematic Review of Therapeutic Potential, Preparation Techniques, and Delivery Methods Across Medical Conditions

Kurdistan Journal of Applied Research, Journal Year: 2024, Volume and Issue: 9(2), P. 65 - 85

Published: Nov. 2, 2024

Fecal microbiota transplantation (FMT) is revolutionizing the treatment of gastrointestinal disorders by leveraging gut microbiome in innovative ways. This systematic review evaluates clinical effectiveness and safety FMT across various medical conditions, offering insights into its therapeutic potential limitations. A comprehensive search PubMed, Web Science, Scopus, Embase, ClinicalTrials.gov from January 2000 to December 2023 identified 97 relevant studies on FMT's efficacy, safety, changes after eliminating duplicates. has demonstrated high success rates, particularly treating recurrent refractory Clostridium difficile infections (CDI), with up 90% effectiveness, establishing it as a primary for antibiotic-resistant cases. FMT’s applications are expanding inflammatory bowel diseases (IBD), including ulcerative colitis Crohn's disease, well metabolic neuropsychiatric conditions. Remission rates IBD range 37-45%, outcomes influenced donor characteristics, stool preparation, disease subtype. mild, self-limiting side effects such transient diarrhea abdominal cramping. However, rare serious adverse events underscore need rigorous screening standardized preparation protocols mitigate risks. ability restore healthy flora highlights promise both systemic management. further research essential establish optimized procedures, guidelines, long-term data facilitate integration mainstream practice.

Language: Английский

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