- Email: [email protected]
93 Long-term Risk of Clostridium difficile Infection Recurrence With or Without Antibiotic Exposure Following Successful Fecal Microbiota Transplant
overall stool frequency and urgency components (0.04
92
Background: Altered gut microbiota due to antibiotic use and host factors has been implicated in increased susceptibility to Clostridium difficile infection (CDI). Gut microbiota signatures associated with CDI have been described but it is unclear if differences in gut microbiota at the time of primary infection predict CDI recurrence after successful treatment. The aim of this study was to identify gut microbiota signatures predictive of recurrent CDI. Methods: Pre-treatment stool samples and clinical outcomes were evaluated from patients with a first episode of CDI. The V4 region of the 16S rRNA gene was used for gut microbial community profiling in stool samples using the MISEQ Illumina platform. Linear discriminant analysis Effect Size (LefSe) analysis was used to detect differences in abundance of microbial taxa with respect to the recurrence groups; followed by a linear discriminant analysis to estimate the effect size of each differentially abundant taxa. Results: Of 88 patients, 77 (87.5%) were treatment responders and responders had a median age of 53.8 years; 59.7% were female. The rate of recurrent CDI in responders was 28.5%. Clinical factors including age, sex, bodymass index, prior antibiotic exposure, acquisition mode, treatment regimen or concomitant systemic antibiotic exposure did not predict recurrent CDI. Compared to non-recurrent CDI, patients with recurrent CDI had an increase in Veillonella species, Enterobacteriaceae, Streptococcus species, Parabacteroides species and Lachnospiraceae in fecal samples at initial diagnosis using LEfSe analysis. A risk index built from this panel of microbes highly differentiated between the groups with a mean score of 0.09 ±0.16 in recurrent CDI and 0.19 ±0.07 in non-recurrent CDI (p = 0.0001). Receiver operating characteristic (ROC) curve analysis showed that this risk index was a strong predictor of recurrent infection, with an area under the curve of 0.85. A risk index threshold of 0.036 or higher predicted recurrence with sensitivity of 82% at a specificity of 95%; an internal leave-one-out cross-validation validated the model (p <0.0001). Conclusions: Gut microbiota signatures identified in pre-treatment stool can predict recurrent CDI after successful response to initial treatment. This may allow early treatment of patients likely to recur with modalities such as fecal microbiota transplantation.
90 Western Diet Induces a Shift in Microbiota Composition Enhancing Susceptibility to Adherent-Invasive E. Coli Infection and Intestinal Inflammation Allison Agus, Jérémy Denizot, Jonathan Thévenot, Sébastien Massier, Pierre Sauvanet, Richard Bonnet, Elisabeth Billard, Nicolas Barnich OBJECTIVE: Recent advances have shown that the abnormal inflammatory response observed in Crohn's disease (CD) involves an interplay between intestinal microbiota, host genetics and environmental factors. The escalating consumption of fat and sugar in Western countries parallels an increased incidence of CD during the latter 20th century. METHODS: The impact of a High-Fat/High-Sugar (HF/HS) diet in mice was evaluated for the (i) gut micro-inflammation (fecal lipocalin-2), (ii) selection of E. coli and impact on intestinal microbiota composition, (iii) susceptibility to a DSS-induced colitis, (iv) concentration of short-chain fatty acids (SCFA) and (v) expression and potential protective effect on gut inflammation of their free fatty acid receptor, G-protein-coupled receptor 43 (GPR43). RESULTS: HF/HS diet increased Lcn-2 level in stools from 5 weeks until 18 weeks of treatment, showing that HF/HS diet creates a specific inflammatory environment in the gut. This was correlated with intestinal mucosa dysbiosis characterized by reduction of bacterial richness, but also by an overgrowth of pro-inflammatory proteobacteria such as E. coli and a decrease in protective bacteria. In addition, the fecal transplantation from HF/HS treated mice to germ-free mice increased susceptibility to Adherent-Invasive Escherichia coli (AIEC) infection. Interestingly, HF/HS diet led to an exacerbation of gut inflammation following DSSinduced colitis, with an increase of DAI, histological score and release of pro-inflammatory cytokines. Moreover, a significantly decrease of SCFA concentrations in fecal samples from mice fed a HF/HS diet compared with mice fed a conventional diet was observed. The expression of SCFA G-protein coupled receptor 43 (GPR43) was reduced in mice treated with a HF/HS diet and reduced in CD patients compared with controls. Interestingly, mice treated with an agonist of GPR43 were protected against DSS-induced colitis. CONCLUSIONS: Western diet creates a low-grade inflammation in the gut with a decrease of protective SCFA producing bacteria, favoring the overgrowth of opportunistic pathogenic E. coli which could aggravate the inflammatory process resulting in chronic inflammation. Moreover, activation of GPR43 receptor pathway could be used as a new strategy to treat CD patients abnormally colonized by AIEC bacteria.
93 Long-term Risk of Clostridium difficile Infection Recurrence With or Without Antibiotic Exposure Following Successful Fecal Microbiota Transplant Monika Fischer, Emmalee Phelps, Rishi Bolla, Margaret Storm, Jessica R. Allegretti Background: The long-term risk of C. difficile infection (CDI) recurrence after a successful fecal microbiota transplant (FMT) with or without exposure to a non-CDI antibiotic in not known. The role of prophylactic concomitant anti-CDI antibiotics (vancomycin, metronidazole or fidaxomicin) or a probiotic in the prevention of CDI recurrence is debated. Methods: Patients with a history of recurrent CDI who underwent successful FMT (defined as resolution of the diarrhea and/or negative stool C. diff by PCR at 8 weeks) were prospectively followed at multiple time points in clinic and/or by phone at two academic medical centers. A questionnaire was used soliciting information about use and type of any non-CDI antibiotics after FMT, underlying infection/condition treated, use and type of concomitant prophylactic anti-CDI antibiotics and/or probiotic and CDI recurrence. Use of anti-CDI and non-CDI antibiotics was confirmed using EMR and pharmacy records. Results: A total of 265 patients were contacted. Of these had 25 died and 89 either did not respond or antibiotic usage could not be confirmed. Among the 152 patients with complete follow-up, 73% (n=111) were females, average age at FMT was 58 yrs (±SD 18.4) and mean duration of post FMT follow-up was 62 weeks (±SD 37.4, range 12-169). 38% (n=58) received a non-CDI antibiotic, and of these 48% (n=28) used non-CDI antibiotics > 1 time. The reason for non-CDI antibiotic use was UTI in 28% (n=16), pneumonia 21% (n=12), sinusitis and/or URI 12% (n=7), skin infection 10% (n=6), surgery 7% (n=4), diverticulitis 5% (n=3), SIBO 5% (n= 3), dental 3% (n=2) and other in 12% (n=7) of patients. The type of non-CDI antibiotics taken were fluoroquinolones by 37% (n=22) of patients, amoxicillin/amoxicillin-clavulanic acid 26% (n=15), cephalosporins 14% (n=8), sulfamethoxazole/trimethoprim 10% (n= 6),doxycycline 8% (n=5), azithromycin 9% (n=5), clindamycin 3% (n=2), aminoglycoside 3% (n=2), carbapenem 3% (n=2), and fosfomycin 2% (n=1). Prophylactic anti-CDI antibiotics were given to 47% (n=27) of patients. This includes: vancomycin 70% (n=19) and metronidazole 30% (n=8). Additionally, 50% (n=29) of patients took concomitant probiotics: saccharomyces boulardii 28% (n=8), kefir 17% (n=5), or other 55% (n=16). The overall CDI recurrence rate was 10.5% (16/152), of these 62.5% (10/16) were related to non-CDI antibiotic. Of all the non-CDI antibiotic users only 17.2% (10/58) developed recurrence of CDI. Conclusion: Following a successful FMT, the long-term rates of CDI recurrence in the setting of non-CDI antibiotic use is low. Larger studies are needed to evaluate the necessity of concomitant anti-CDI antibiotics and probiotics in order to prevent CDI recurrence in the setting of non-CDI antibiotic use.
91 Mass Spectrometry Imaging (MSI) of Microbiome-Metabolome Interactions in Colorectal Cancer James L. Alexander, Alasdair Scott, Anna Mroz, Alvaro Perdones-Montero, James Mckenzie, Douglas N. Rees, Abigail Speller, Kirill Veselkov, James M. Kinross, Zoltan Takats, Julian Marchesi, Julian P. Teare Introduction The mucosal colorectal cancer microbiome is highly personalised and niche specific. Nevertheless, microbial co-metabolites such as bile acids and short chain fatty acids are established modifiers of cancer risk. Novel approaches are required for spatially resolved analysis of the metabolic function of the cancer microbiome. Methods A prospective, observational multi-centred study was performed in a single cohort of patients undergoing elective surgery for colorectal tumors at three London teaching hospitals. Tissue from on tumor, and from normal mucosa at 10cms were sampled and snap frozen at -80°C. DNA was extracted from mucosal samples using PowerLyzer PowerSoil DNA Isolation Kit (MO BIO) and the V1-2 regions of the 16S rRNA gene sequences were sequenced on the Illumina MiSeq platform. Data were analysed by Mothur to determine OTU abundance per sample and were further analysed in Stamp and R. Metabolomic analysis was performed on corresponding tissue from the same samples by Desorption Electrospray Ionisation Imaging Mass Spectrometry (DESI-MSI) for spatially resolved chemical analysis of microbiome biochemistry, and multivariate analysis was performed in Matlab using in house protocols. Results 46 patients with sporadic colorectal cancer were recruited (M:F 23:23, median age 69, range 35-89). Tumor locations were 16 rectal, 11 sigmoid, 5 transverse, 14 caecum & ascending colon. Cancers were staged as 1 T1, 6 T2, 27 T3 & 12 T4; 26 N0, 14 N1, 6 N2; 34 M0, 12 M1/ 2. No difference was seen between on and off tumor sites in Chao richness or Shannon diversity indices. Fusobacterium (10/46 patients) and Campylobacter genuses (11/46) were overexpressed on tumors compared to adjacent normal mucosa (p=0.007); Blautia, Colinsella and Staphylococcus were overexpressed on adjacent normal mucosa (p=0.024, p=0.025 & p=0.028). Inter-individual taxonomic variation out-weighed OTU enrichments between tissue types. Metabolomic analysis using DESI-MSI demonstrated that the cancer lipidome harbors topographically discrete regions of lipids that are highly diagnostic for colon cancer: Phosphatidylinositol (38:3) and Phophatidylethanolamine (-32:1) were preferentially found in cancer and Phosphatidic Acid (36:2) in normal tissue (p<0.001). DESI-MSI was also able to visualise topographically discrete microbial co-metabolites such as bile acids within tumors. Conclusion Inter-individual variation significantly influences the mucosal expression of pathobionts currently linked to colorectal cancer. However, this analysis suggests that mutualistic community metabolism within geographically discrete regions of tumors is likely to be of functional importance to cancer aetiology.
94 Probiotic Saccharomyces boulardii CNCM I-745 Inhibits Hypervirulent Clostridium difficile-Associated Cecal Tissue Damage and Pro-Inflammatory Cytokine Expression in Hamsters Hon Wai Koon, Diana Hoang-Ngoc Tran, Xinhua Chen, Ciaran P. Kelly, Charalabos Pothoulakis Background and Aims: Clostridium difficile infection (CDI) is a common debilitating nosocomial infection associated with high morbidity,mortality, and increasing incidence. Several severe CDI outbreaks have been reported, and many of these are associated with hypervirulent strains, including ribotypes 027, 017, and 078. Clinical and experimental evidence indicates that the non-pathogenic yeast Saccharomyces boulardii CNCM I-745 (S.b) is effective for the prevention and treatment of CDI. However, there is no evidence suggesting that this probiotic can protect CDI caused by hypervirulent C. difficile strains. Materials and Methods: We
S-23
AGA Abstracts
AGA Abstracts
Gut Microbiome Signatures at the Time of Primary Clostridium difficile Infection Predict Recurrence Sahil Khanna, Emmanuel Montassier, Robin Patel, Patricia P. Kammer, Dan Knights, Darrell Pardi, Purna C. Kashyap
92
Background: Altered gut microbiota due to antibiotic use and host factors has been implicated in increased susceptibility to Clostridium difficile infection (CDI). Gut microbiota signatures associated with CDI have been described but it is unclear if differences in gut microbiota at the time of primary infection predict CDI recurrence after successful treatment. The aim of this study was to identify gut microbiota signatures predictive of recurrent CDI. Methods: Pre-treatment stool samples and clinical outcomes were evaluated from patients with a first episode of CDI. The V4 region of the 16S rRNA gene was used for gut microbial community profiling in stool samples using the MISEQ Illumina platform. Linear discriminant analysis Effect Size (LefSe) analysis was used to detect differences in abundance of microbial taxa with respect to the recurrence groups; followed by a linear discriminant analysis to estimate the effect size of each differentially abundant taxa. Results: Of 88 patients, 77 (87.5%) were treatment responders and responders had a median age of 53.8 years; 59.7% were female. The rate of recurrent CDI in responders was 28.5%. Clinical factors including age, sex, bodymass index, prior antibiotic exposure, acquisition mode, treatment regimen or concomitant systemic antibiotic exposure did not predict recurrent CDI. Compared to non-recurrent CDI, patients with recurrent CDI had an increase in Veillonella species, Enterobacteriaceae, Streptococcus species, Parabacteroides species and Lachnospiraceae in fecal samples at initial diagnosis using LEfSe analysis. A risk index built from this panel of microbes highly differentiated between the groups with a mean score of 0.09 ±0.16 in recurrent CDI and 0.19 ±0.07 in non-recurrent CDI (p = 0.0001). Receiver operating characteristic (ROC) curve analysis showed that this risk index was a strong predictor of recurrent infection, with an area under the curve of 0.85. A risk index threshold of 0.036 or higher predicted recurrence with sensitivity of 82% at a specificity of 95%; an internal leave-one-out cross-validation validated the model (p <0.0001). Conclusions: Gut microbiota signatures identified in pre-treatment stool can predict recurrent CDI after successful response to initial treatment. This may allow early treatment of patients likely to recur with modalities such as fecal microbiota transplantation.
90 Western Diet Induces a Shift in Microbiota Composition Enhancing Susceptibility to Adherent-Invasive E. Coli Infection and Intestinal Inflammation Allison Agus, Jérémy Denizot, Jonathan Thévenot, Sébastien Massier, Pierre Sauvanet, Richard Bonnet, Elisabeth Billard, Nicolas Barnich OBJECTIVE: Recent advances have shown that the abnormal inflammatory response observed in Crohn's disease (CD) involves an interplay between intestinal microbiota, host genetics and environmental factors. The escalating consumption of fat and sugar in Western countries parallels an increased incidence of CD during the latter 20th century. METHODS: The impact of a High-Fat/High-Sugar (HF/HS) diet in mice was evaluated for the (i) gut micro-inflammation (fecal lipocalin-2), (ii) selection of E. coli and impact on intestinal microbiota composition, (iii) susceptibility to a DSS-induced colitis, (iv) concentration of short-chain fatty acids (SCFA) and (v) expression and potential protective effect on gut inflammation of their free fatty acid receptor, G-protein-coupled receptor 43 (GPR43). RESULTS: HF/HS diet increased Lcn-2 level in stools from 5 weeks until 18 weeks of treatment, showing that HF/HS diet creates a specific inflammatory environment in the gut. This was correlated with intestinal mucosa dysbiosis characterized by reduction of bacterial richness, but also by an overgrowth of pro-inflammatory proteobacteria such as E. coli and a decrease in protective bacteria. In addition, the fecal transplantation from HF/HS treated mice to germ-free mice increased susceptibility to Adherent-Invasive Escherichia coli (AIEC) infection. Interestingly, HF/HS diet led to an exacerbation of gut inflammation following DSSinduced colitis, with an increase of DAI, histological score and release of pro-inflammatory cytokines. Moreover, a significantly decrease of SCFA concentrations in fecal samples from mice fed a HF/HS diet compared with mice fed a conventional diet was observed. The expression of SCFA G-protein coupled receptor 43 (GPR43) was reduced in mice treated with a HF/HS diet and reduced in CD patients compared with controls. Interestingly, mice treated with an agonist of GPR43 were protected against DSS-induced colitis. CONCLUSIONS: Western diet creates a low-grade inflammation in the gut with a decrease of protective SCFA producing bacteria, favoring the overgrowth of opportunistic pathogenic E. coli which could aggravate the inflammatory process resulting in chronic inflammation. Moreover, activation of GPR43 receptor pathway could be used as a new strategy to treat CD patients abnormally colonized by AIEC bacteria.
93 Long-term Risk of Clostridium difficile Infection Recurrence With or Without Antibiotic Exposure Following Successful Fecal Microbiota Transplant Monika Fischer, Emmalee Phelps, Rishi Bolla, Margaret Storm, Jessica R. Allegretti Background: The long-term risk of C. difficile infection (CDI) recurrence after a successful fecal microbiota transplant (FMT) with or without exposure to a non-CDI antibiotic in not known. The role of prophylactic concomitant anti-CDI antibiotics (vancomycin, metronidazole or fidaxomicin) or a probiotic in the prevention of CDI recurrence is debated. Methods: Patients with a history of recurrent CDI who underwent successful FMT (defined as resolution of the diarrhea and/or negative stool C. diff by PCR at 8 weeks) were prospectively followed at multiple time points in clinic and/or by phone at two academic medical centers. A questionnaire was used soliciting information about use and type of any non-CDI antibiotics after FMT, underlying infection/condition treated, use and type of concomitant prophylactic anti-CDI antibiotics and/or probiotic and CDI recurrence. Use of anti-CDI and non-CDI antibiotics was confirmed using EMR and pharmacy records. Results: A total of 265 patients were contacted. Of these had 25 died and 89 either did not respond or antibiotic usage could not be confirmed. Among the 152 patients with complete follow-up, 73% (n=111) were females, average age at FMT was 58 yrs (±SD 18.4) and mean duration of post FMT follow-up was 62 weeks (±SD 37.4, range 12-169). 38% (n=58) received a non-CDI antibiotic, and of these 48% (n=28) used non-CDI antibiotics > 1 time. The reason for non-CDI antibiotic use was UTI in 28% (n=16), pneumonia 21% (n=12), sinusitis and/or URI 12% (n=7), skin infection 10% (n=6), surgery 7% (n=4), diverticulitis 5% (n=3), SIBO 5% (n= 3), dental 3% (n=2) and other in 12% (n=7) of patients. The type of non-CDI antibiotics taken were fluoroquinolones by 37% (n=22) of patients, amoxicillin/amoxicillin-clavulanic acid 26% (n=15), cephalosporins 14% (n=8), sulfamethoxazole/trimethoprim 10% (n= 6),doxycycline 8% (n=5), azithromycin 9% (n=5), clindamycin 3% (n=2), aminoglycoside 3% (n=2), carbapenem 3% (n=2), and fosfomycin 2% (n=1). Prophylactic anti-CDI antibiotics were given to 47% (n=27) of patients. This includes: vancomycin 70% (n=19) and metronidazole 30% (n=8). Additionally, 50% (n=29) of patients took concomitant probiotics: saccharomyces boulardii 28% (n=8), kefir 17% (n=5), or other 55% (n=16). The overall CDI recurrence rate was 10.5% (16/152), of these 62.5% (10/16) were related to non-CDI antibiotic. Of all the non-CDI antibiotic users only 17.2% (10/58) developed recurrence of CDI. Conclusion: Following a successful FMT, the long-term rates of CDI recurrence in the setting of non-CDI antibiotic use is low. Larger studies are needed to evaluate the necessity of concomitant anti-CDI antibiotics and probiotics in order to prevent CDI recurrence in the setting of non-CDI antibiotic use.
91 Mass Spectrometry Imaging (MSI) of Microbiome-Metabolome Interactions in Colorectal Cancer James L. Alexander, Alasdair Scott, Anna Mroz, Alvaro Perdones-Montero, James Mckenzie, Douglas N. Rees, Abigail Speller, Kirill Veselkov, James M. Kinross, Zoltan Takats, Julian Marchesi, Julian P. Teare Introduction The mucosal colorectal cancer microbiome is highly personalised and niche specific. Nevertheless, microbial co-metabolites such as bile acids and short chain fatty acids are established modifiers of cancer risk. Novel approaches are required for spatially resolved analysis of the metabolic function of the cancer microbiome. Methods A prospective, observational multi-centred study was performed in a single cohort of patients undergoing elective surgery for colorectal tumors at three London teaching hospitals. Tissue from on tumor, and from normal mucosa at 10cms were sampled and snap frozen at -80°C. DNA was extracted from mucosal samples using PowerLyzer PowerSoil DNA Isolation Kit (MO BIO) and the V1-2 regions of the 16S rRNA gene sequences were sequenced on the Illumina MiSeq platform. Data were analysed by Mothur to determine OTU abundance per sample and were further analysed in Stamp and R. Metabolomic analysis was performed on corresponding tissue from the same samples by Desorption Electrospray Ionisation Imaging Mass Spectrometry (DESI-MSI) for spatially resolved chemical analysis of microbiome biochemistry, and multivariate analysis was performed in Matlab using in house protocols. Results 46 patients with sporadic colorectal cancer were recruited (M:F 23:23, median age 69, range 35-89). Tumor locations were 16 rectal, 11 sigmoid, 5 transverse, 14 caecum & ascending colon. Cancers were staged as 1 T1, 6 T2, 27 T3 & 12 T4; 26 N0, 14 N1, 6 N2; 34 M0, 12 M1/ 2. No difference was seen between on and off tumor sites in Chao richness or Shannon diversity indices. Fusobacterium (10/46 patients) and Campylobacter genuses (11/46) were overexpressed on tumors compared to adjacent normal mucosa (p=0.007); Blautia, Colinsella and Staphylococcus were overexpressed on adjacent normal mucosa (p=0.024, p=0.025 & p=0.028). Inter-individual taxonomic variation out-weighed OTU enrichments between tissue types. Metabolomic analysis using DESI-MSI demonstrated that the cancer lipidome harbors topographically discrete regions of lipids that are highly diagnostic for colon cancer: Phosphatidylinositol (38:3) and Phophatidylethanolamine (-32:1) were preferentially found in cancer and Phosphatidic Acid (36:2) in normal tissue (p<0.001). DESI-MSI was also able to visualise topographically discrete microbial co-metabolites such as bile acids within tumors. Conclusion Inter-individual variation significantly influences the mucosal expression of pathobionts currently linked to colorectal cancer. However, this analysis suggests that mutualistic community metabolism within geographically discrete regions of tumors is likely to be of functional importance to cancer aetiology.
94 Probiotic Saccharomyces boulardii CNCM I-745 Inhibits Hypervirulent Clostridium difficile-Associated Cecal Tissue Damage and Pro-Inflammatory Cytokine Expression in Hamsters Hon Wai Koon, Diana Hoang-Ngoc Tran, Xinhua Chen, Ciaran P. Kelly, Charalabos Pothoulakis Background and Aims: Clostridium difficile infection (CDI) is a common debilitating nosocomial infection associated with high morbidity,mortality, and increasing incidence. Several severe CDI outbreaks have been reported, and many of these are associated with hypervirulent strains, including ribotypes 027, 017, and 078. Clinical and experimental evidence indicates that the non-pathogenic yeast Saccharomyces boulardii CNCM I-745 (S.b) is effective for the prevention and treatment of CDI. However, there is no evidence suggesting that this probiotic can protect CDI caused by hypervirulent C. difficile strains. Materials and Methods: We
S-23
AGA Abstracts
AGA Abstracts
Gut Microbiome Signatures at the Time of Primary Clostridium difficile Infection Predict Recurrence Sahil Khanna, Emmanuel Montassier, Robin Patel, Patricia P. Kammer, Dan Knights, Darrell Pardi, Purna C. Kashyap