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Su1741 Psychological Stress Exacerbates Indomethacin-Induced Small Intestinal Injury Possibly via Changes in Intestinal Microbiota
AGA Abstracts
Figure 1: OTUs plotted from least ubiquitous (top) to most ubiquitous (bottom). Darker colors indicate higher abundance.
Figure 1. Relative abundance of Proteobacteria after FMT
Figure 2: The same plot as in Figure 1, with only the two OTUs most enriched before FMT samples and three OTUs most enriched following FMT shown.
Su1740 Dynamic Changes in the Intestinal Microbiota Following Fecal Microbiota Transplantation for Refractory Inflammatory Bowel Disease in Children Brian R. Bush, Matthew B. Rogers, Brian Firek, Adam Kufen, Zachary Jackson, Michael Morowitz, Alka Goyal Background: Inflammatory bowel disease (IBD) has been associated with dysbiosis. Fecal microbiota transplantation (FMT) has been demonstrated to restore gut bacterial diversity in patients with recurrent Clostridium difficile infection (CDI). FMT is still an investigational treatment for IBD. Little is known about the stability of fecal microbiota following FMT in IBD patients. We report on short- and long-term changes in the composition of the fecal microbiota in children undergoing FMT for refractory colitis secondary to Ulcerative colitis (UC) or Crohn's Disease (CD). Methodology: Children with mild-moderately active, refractory UC or CD (CDI negative) were recruited for study participation with institutional approval. Stool samples were collected from the donor and the patient prior to FMT. Followup stool samples from patients were collected at 1 week, 1 month, and 6 months. Bacterial DNA was extracted and 16S rRNA gene sequences were PCR amplified from the stool. Sequencing data thus obtained with the Illumina MiSeq platform were analyzed with QIIME software for taxonomic assignments and calculations of microbial diversity. LEfSE was used to compare healthy volunteers in the Human Microbiome Project (HMP). Disease activity before and after FMT was measured using Pediatric Crohn's Disease Activity Index (PCDAI) and Pediatric Ulcerative Colitis Activity Index (PUCAI) scores. Results: Ten patients with a primary diagnosis of UC (n=8) or CD (n=2) were enrolled and underwent FMT. The mean age of subjects was 11.8 y (range 8-21 y). Pre-FMT PUCAI scores ranged from 20-55, and PCDAI was 5-12.5. Relative to HMP samples and donor samples, pre-FMT IBD samples were highly enriched with Proteobacteria (family Enterobacteriaceae) and Fusobacteria and depleted of strict anaerobes from phylum Bacteroides. Following FMT, the abundance of Proteobacteria decreased significantly (Figure 1) as the fecal microbiota shifted rapidly to a donor-like state in 6 of 10 patients. A heatmap of Pearson correlation coefficients (Figure 2) demonstrates that the microbiota of 6 patients appeared highly similar to donor samples at 1 week, but the composition diverged on an individual basis at later time points. In several cases, dysbiosis reappeared within 6 month samples. Clinical response defined as a drop by 12.5 points in PCDAI and 15 points in PUCAI was noted in 70%, 60%, and 22% of study subjects at 1 week, 1 month and 6 months respectively. Most of these responses corresponded to successful modification of the fecal microbiota with FMT. Two subjects had no demonstrable clinical response. Conclusions: We conclude that FMT is a potential therapeutic option to treat refractory pediatric IBD by modifying gut bacterial populations. Future treatment regimens may test the efficacy of multiple serial FMT procedures and may be guided by real-time monitoring of the gut microbiota.
Figure 2. Pearson Correlation Coefficients
Su1741 Psychological Stress Exacerbates Indomethacin-Induced Small Intestinal Injury Possibly via Changes in Intestinal Microbiota Kenichi Yoshikawa, Chie Kurihara, Hirotaka Furuhashi, Takeshi Takajo, Koji Maruta, Yuichi Yasutake, Yoshikiyo Okada, Masaaki Higashiyama, Chikako Watanabe, Shunsuke Komoto, Kengo Tomita, Shigeaki Nagao, Soichiro Miura, Ryota Hokari Background: Nonsteroidal anti-inflammatory drugs (NSAIDs) induce intestinal lesions frequently than we expected previously. However, little is known whether psychological stress affects the small intestinal injury induced by NSAIDs (NSAIDs enteropathy). Recently it is well known that psychological stress may exacerbate various gastrointestinal diseases. In this study effect of psychological stress on NSAIDs enteropathy was investigated by using repeated water avoidance stress (rWAS) model and we further investigated whether NSAIDs enteropathy could be aggravated by transplantation of microbiota from stress-laden mice. Methods: Experiment1: Mice were treated rWAS for 1 hr for 8 days. Enteropathy was induced by indomethacin (IND) injection (10mg/kg) subcutaneously at the final day of rWAS. Mice were divided into 4 groups for stress/IND; (-)/(-), (-)/(+), (+)/(-), and (+)/(+). The total areas of ulcers were measured after Evans-blue injection, histological damage was assessed, and intestinal tissue TNFa expression was measured by qRT-PCR. Intestinal permeability was measured fluorescence intensity 1hr after FITC-dextran administration, and microbial community in distal ileum were analyzed by RT-PCR. Experiment2: 2 groups of mice were administered antibiotics cocktail for 4 weeks to deplete microbiota, then cecal microbiota transplantation was conducted with 5 days' administration orally of cecal contents from stress(-) or (+) mice, and then enteropathy was induced by IND without rWAS treatment. Results: Experiment1: Stress(+)IND(+) mice significantly exacerbated macroscopic small intestinal injury (25.9±1.1 vs 21.4±1.3 mm2) and histological severity score compared to stress(-)IND(+) group. Stress(+) mice also significantly increased levels of tissue TNFa and intestinal permeability. Gut microbiota community analysis showed increase of total bacterial number in stress(+) group 6 times compared to stress(-) group, particularly in Bacteroidetes (146 times) and Actinobacteria (91 times) for phylum level. Abundance ratio of Firmicutes and Bacteroidetes changed from 95% and 2% in stress(-) to 79% and 19% in stress(+), respectively. Experiment2: In mice administered from cecal contents of stress(+) mice, the intestinal injury (10±4.5 vs 6.6±1.6 mm2), tissue TNFa expression, and intestinal permeability were significantly greater than those from stress(-) mice. Conclusion: Our results indicate the possible involvement of brain-gut axis in the pathophysiology of NSAIDs enteropathy.
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AGA Abstracts
AGA Abstracts
Susceptibility to NSAIDs and intestinal permeability enhancement by psychological stress were successfully transferred via cecal microbiota transplantation. Although effective treatment for NSAIDs enteropathy has not been established, modification of intestinal microbiota is suggested as a possible target for its treatment especially under psychological stress.
Su1742 Fecal Microbiota Transplantation for Recurrent Clostridium difficile Infection in Patients with Ileal Pouches Nan Lan, Luca Stocchi, Feza H. Remzi, Bo Shen Introductions: Clostridium difficile (C. difficile) infection (CDI) in patients underwent ileal pouch-anal anastomosis (IPAA) has been increasingly recognized. There is only one case report in which fecal microbiota transplantation (FMT) was shown to be effective in treating CDI of the pouch. The aim of this study was to report the largest series of ileal pouch patients with recurrent CDI that was treated by FMT. Methods: All patients underwent FMT for CDI were extracted from our IRB-approved, prospectively maintained Registry of Pouch Disorders. Recurrent CDI were defined as PCR confirmed CDI diagnosis with proper antibiotic treatments. Donors were screened with our institution-based standardized protocol. Antegrade and/or retrograde FMT were performed were recorded in detail. The primary outcome was more than one negative PCR results for C. diffcile after FMT and the secondary outcomes were symptomatic and endoscopic response and procedure-associated complications. Results: A total of 10 patients were included in this study and all of them were Caucasian males. All patients had a preoperative diagnosis of extensive ulcerative colitis with colectomy indications of medically refractory disease (N = 8) or colitis-associated neoplasia (N = 2). Before FMT, these patients had a mean of 2.1±1.0 courses of antibiotic treatments including oral vancomycin, metronidazole or fidaxomicin. A total of 18 sessions of FMT were given to the 10 patients with an average of 1.8±1.2 sessions each. Within the 18 sessions, 12 were given via pouchoscopy, 4 via EGD and 2 via enema. For those given via pouchoscopy, a mean of 179.2± 61.2 ml of saline-diluted stool were administered and those given through EGD, 30 ml of the same solute were administered. All patients tested negative on C. difficile PCR after FMT. During a mean follow-up of 1.8±1.2 years, a total of 6 (66.7%) patients showed significant symptomatic improvements, while 3 (33.3%) showed none and 1 lost follow-up. Though effective symptomatically, the Pouchitis Disease Activity Index endoscopy subscores showed a significant improvement in only 2 patients, while no improvement in 5 patients and 3 patients had no follow-up pouchoscopy (Figure 1). There were one patient that had one recurrence after initial successful treatment and another that had two. They were both treated again with FMT successfully. One patient experienced nausea and mild abdominal pain after the treatment, presumably due to ileus. He was hospitalized and treated conservatively. A total of two patients had pouch failure. One was due to bowel obstruction, while the other due to ischemic pouchitis. Conclusions: Our findings suggest that FMT appeared to be a potent alternative for treating recurrent CDI in patients with ileal pouches. Though FMT was effective in the eradication of CDI, the improvement observed on endoscopy was minimal. Table 1. Patient's Clinical Characteristics (N=10).
Su1743 Characterization of Fecal Microbiota in Response to Heterologous Versus Autologous (Placebo) Fecal Microbial Transplantation: Results From a DualCenter, Randomized, Placebo-Controlled Trial Christopher Staley, Colleen R. Kelly, Lawrence J. Brandt, Alexander Khoruts, Michael J. Sadowsky Background: Fecal microbial transplantation (FMT) has been increasingly used as an alternative treatment for recurrent Clostridium difficile infections (RDCI), with a success rate averaging 90%. This treatment has recently been validated in a dual-center, blinded, randomized, placebo-controlled clinical trial, testing the efficacy and safety of healthy donor stool (heterologous FMT) compared with autologous stool, (placebo FMT). Overall, heterologous stool was significantly more effective than autologous (placebo) stool (91% vs 63%), but of interest was the notably higher success rate for autologous FMT observed at the Bronx, NY site (90%) compared with the Providence, RI site (43%). Objective: The fecal microbial compositions of heterologous and autologous FMT recipients were characterized to assess any differences in community composition resulting from fecal material used, pre-FMT treatments, or patient demographic factors. Methods: The fecal microbiome of patients was characterized using Illumina next-generation sequencing of the V5+V6 hypervariable regions of the 16S rRNA gene. Patient samples were collected at baseline, at least 5 days prior to FMT, and again at 2 and 8 weeks post-FMT. Results: Both pre-FMT patient and donor bacterial communities differed significantly at the 2 institutions (P < 0.001). Bacterial communities prior to FMT were dysbiotic and characterized at the phylum level by drastic reduction or disappearance of Bacteroidetes and expansion of the Proteobacteria and/or Verrucomicrobia. Abundances of Verrucomicrobia and Bacteroidetes in pre-FMT samples were notably greater at the Bronx site, where greater autologous FMT success was observed. Heterologous FMT restored the bacterial community alpha diversity to resemble that of healthy donors, while the bacterial community composition in fecal samples of patients also shifted to one similar to donor samples. Community composition after autologous FMT was significantly different than that resulting from heterologous FMT at both sites (P £0.025), and the abundance of Bacteroidetes did not recover to the same extent as was observed in patients who received heterologous donor material. Conclusions: These data reveal that autologous FMT results in different community composition than heterologous FMT, with the latter providing faster and more marked resolution of dysbiotic microbial signatures. Site-specific parameters including antiC. difficile treatment regimens as well as other parameters (e.g. numbers of recurrences and use of proton pump inhibitors) may be driving differences in community composition between centers and subsequent treatment success. Taken together, these results suggest that pre-FMT alteration in the microbiota by anti- C. difficile treatment, for example, may influence the necessity for or success of FMT therapy.
Su1744 Lyophilized Fecal Microbiota Transplantation and Cryoprotectants for Viable Bacteria Preservation Scott W. Mitchell, Anthony Jaworski, Michelle Bull, Connie Wong, Vanessa Furnari, Belinda Chapman, Thomas Borody Introduction: The use of cryoprotectants in foodstuffs, medications and biologic materials has considerable history. However, there is minimal data on the use of cryoprotectants to preserve viable bacteria within lyophilized Fecal Microbiota Transplantation (FMT) product. FMT has received considerable attention for its superior treatment of recurrent Clostridium difficile infection (CDI) compared with antibiotics1. Lyophilized fecal material can be stored for much longer than liquefied stool. The ability to store FMT material for longer relieves the time pressure associated with completion of donor pathology screening to ensure FMT safety. Success of FMT using lyophilized material will depend on preservation of bacterial viability. The choice of cryoprotectant may be important in the optimisation of bacterial viability in lyophilized FMT product. Aim: To measure the total live cell count and compare proportional viability of microorganisms in reconstituted lyophilized FMT following lyophilization using a range of cryoprotectants. Methods: FMT was prepared at a ratio of 1:4 donor stool to saline diluent containing various cryoprotectants. Following homogenization nonbacterial component removal and product concentration were achieved with 250µm filter bags and a refrigerated centrifuge. The resulting slurry was then lyophilized (final water activity £ 0.3). Total and total viable bacterial counts were determined immediately after lyophilization using flow cytometry and Bacteria Count and Live/Dead BacLight ™ kits (Invitrogen). All cryoprotectant formulations were tested at least in triplicate (n = 3 - 6). Total live cell counts were normalised among experiments by expressing the results as the proportion (%) of total live cells. One way analysis of variance was used to compare total live cell proportions among the cryoprotectant formulations. Results: Total live cell proportions immediately following lyophilization ranged from 28.5 - 39.0% (see Table).
AGA Abstracts
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Figure 1: OTUs plotted from least ubiquitous (top) to most ubiquitous (bottom). Darker colors indicate higher abundance.
Figure 1. Relative abundance of Proteobacteria after FMT
Figure 2: The same plot as in Figure 1, with only the two OTUs most enriched before FMT samples and three OTUs most enriched following FMT shown.
Su1740 Dynamic Changes in the Intestinal Microbiota Following Fecal Microbiota Transplantation for Refractory Inflammatory Bowel Disease in Children Brian R. Bush, Matthew B. Rogers, Brian Firek, Adam Kufen, Zachary Jackson, Michael Morowitz, Alka Goyal Background: Inflammatory bowel disease (IBD) has been associated with dysbiosis. Fecal microbiota transplantation (FMT) has been demonstrated to restore gut bacterial diversity in patients with recurrent Clostridium difficile infection (CDI). FMT is still an investigational treatment for IBD. Little is known about the stability of fecal microbiota following FMT in IBD patients. We report on short- and long-term changes in the composition of the fecal microbiota in children undergoing FMT for refractory colitis secondary to Ulcerative colitis (UC) or Crohn's Disease (CD). Methodology: Children with mild-moderately active, refractory UC or CD (CDI negative) were recruited for study participation with institutional approval. Stool samples were collected from the donor and the patient prior to FMT. Followup stool samples from patients were collected at 1 week, 1 month, and 6 months. Bacterial DNA was extracted and 16S rRNA gene sequences were PCR amplified from the stool. Sequencing data thus obtained with the Illumina MiSeq platform were analyzed with QIIME software for taxonomic assignments and calculations of microbial diversity. LEfSE was used to compare healthy volunteers in the Human Microbiome Project (HMP). Disease activity before and after FMT was measured using Pediatric Crohn's Disease Activity Index (PCDAI) and Pediatric Ulcerative Colitis Activity Index (PUCAI) scores. Results: Ten patients with a primary diagnosis of UC (n=8) or CD (n=2) were enrolled and underwent FMT. The mean age of subjects was 11.8 y (range 8-21 y). Pre-FMT PUCAI scores ranged from 20-55, and PCDAI was 5-12.5. Relative to HMP samples and donor samples, pre-FMT IBD samples were highly enriched with Proteobacteria (family Enterobacteriaceae) and Fusobacteria and depleted of strict anaerobes from phylum Bacteroides. Following FMT, the abundance of Proteobacteria decreased significantly (Figure 1) as the fecal microbiota shifted rapidly to a donor-like state in 6 of 10 patients. A heatmap of Pearson correlation coefficients (Figure 2) demonstrates that the microbiota of 6 patients appeared highly similar to donor samples at 1 week, but the composition diverged on an individual basis at later time points. In several cases, dysbiosis reappeared within 6 month samples. Clinical response defined as a drop by 12.5 points in PCDAI and 15 points in PUCAI was noted in 70%, 60%, and 22% of study subjects at 1 week, 1 month and 6 months respectively. Most of these responses corresponded to successful modification of the fecal microbiota with FMT. Two subjects had no demonstrable clinical response. Conclusions: We conclude that FMT is a potential therapeutic option to treat refractory pediatric IBD by modifying gut bacterial populations. Future treatment regimens may test the efficacy of multiple serial FMT procedures and may be guided by real-time monitoring of the gut microbiota.
Figure 2. Pearson Correlation Coefficients
Su1741 Psychological Stress Exacerbates Indomethacin-Induced Small Intestinal Injury Possibly via Changes in Intestinal Microbiota Kenichi Yoshikawa, Chie Kurihara, Hirotaka Furuhashi, Takeshi Takajo, Koji Maruta, Yuichi Yasutake, Yoshikiyo Okada, Masaaki Higashiyama, Chikako Watanabe, Shunsuke Komoto, Kengo Tomita, Shigeaki Nagao, Soichiro Miura, Ryota Hokari Background: Nonsteroidal anti-inflammatory drugs (NSAIDs) induce intestinal lesions frequently than we expected previously. However, little is known whether psychological stress affects the small intestinal injury induced by NSAIDs (NSAIDs enteropathy). Recently it is well known that psychological stress may exacerbate various gastrointestinal diseases. In this study effect of psychological stress on NSAIDs enteropathy was investigated by using repeated water avoidance stress (rWAS) model and we further investigated whether NSAIDs enteropathy could be aggravated by transplantation of microbiota from stress-laden mice. Methods: Experiment1: Mice were treated rWAS for 1 hr for 8 days. Enteropathy was induced by indomethacin (IND) injection (10mg/kg) subcutaneously at the final day of rWAS. Mice were divided into 4 groups for stress/IND; (-)/(-), (-)/(+), (+)/(-), and (+)/(+). The total areas of ulcers were measured after Evans-blue injection, histological damage was assessed, and intestinal tissue TNFa expression was measured by qRT-PCR. Intestinal permeability was measured fluorescence intensity 1hr after FITC-dextran administration, and microbial community in distal ileum were analyzed by RT-PCR. Experiment2: 2 groups of mice were administered antibiotics cocktail for 4 weeks to deplete microbiota, then cecal microbiota transplantation was conducted with 5 days' administration orally of cecal contents from stress(-) or (+) mice, and then enteropathy was induced by IND without rWAS treatment. Results: Experiment1: Stress(+)IND(+) mice significantly exacerbated macroscopic small intestinal injury (25.9±1.1 vs 21.4±1.3 mm2) and histological severity score compared to stress(-)IND(+) group. Stress(+) mice also significantly increased levels of tissue TNFa and intestinal permeability. Gut microbiota community analysis showed increase of total bacterial number in stress(+) group 6 times compared to stress(-) group, particularly in Bacteroidetes (146 times) and Actinobacteria (91 times) for phylum level. Abundance ratio of Firmicutes and Bacteroidetes changed from 95% and 2% in stress(-) to 79% and 19% in stress(+), respectively. Experiment2: In mice administered from cecal contents of stress(+) mice, the intestinal injury (10±4.5 vs 6.6±1.6 mm2), tissue TNFa expression, and intestinal permeability were significantly greater than those from stress(-) mice. Conclusion: Our results indicate the possible involvement of brain-gut axis in the pathophysiology of NSAIDs enteropathy.
S-541
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AGA Abstracts
AGA Abstracts
Susceptibility to NSAIDs and intestinal permeability enhancement by psychological stress were successfully transferred via cecal microbiota transplantation. Although effective treatment for NSAIDs enteropathy has not been established, modification of intestinal microbiota is suggested as a possible target for its treatment especially under psychological stress.
Su1742 Fecal Microbiota Transplantation for Recurrent Clostridium difficile Infection in Patients with Ileal Pouches Nan Lan, Luca Stocchi, Feza H. Remzi, Bo Shen Introductions: Clostridium difficile (C. difficile) infection (CDI) in patients underwent ileal pouch-anal anastomosis (IPAA) has been increasingly recognized. There is only one case report in which fecal microbiota transplantation (FMT) was shown to be effective in treating CDI of the pouch. The aim of this study was to report the largest series of ileal pouch patients with recurrent CDI that was treated by FMT. Methods: All patients underwent FMT for CDI were extracted from our IRB-approved, prospectively maintained Registry of Pouch Disorders. Recurrent CDI were defined as PCR confirmed CDI diagnosis with proper antibiotic treatments. Donors were screened with our institution-based standardized protocol. Antegrade and/or retrograde FMT were performed were recorded in detail. The primary outcome was more than one negative PCR results for C. diffcile after FMT and the secondary outcomes were symptomatic and endoscopic response and procedure-associated complications. Results: A total of 10 patients were included in this study and all of them were Caucasian males. All patients had a preoperative diagnosis of extensive ulcerative colitis with colectomy indications of medically refractory disease (N = 8) or colitis-associated neoplasia (N = 2). Before FMT, these patients had a mean of 2.1±1.0 courses of antibiotic treatments including oral vancomycin, metronidazole or fidaxomicin. A total of 18 sessions of FMT were given to the 10 patients with an average of 1.8±1.2 sessions each. Within the 18 sessions, 12 were given via pouchoscopy, 4 via EGD and 2 via enema. For those given via pouchoscopy, a mean of 179.2± 61.2 ml of saline-diluted stool were administered and those given through EGD, 30 ml of the same solute were administered. All patients tested negative on C. difficile PCR after FMT. During a mean follow-up of 1.8±1.2 years, a total of 6 (66.7%) patients showed significant symptomatic improvements, while 3 (33.3%) showed none and 1 lost follow-up. Though effective symptomatically, the Pouchitis Disease Activity Index endoscopy subscores showed a significant improvement in only 2 patients, while no improvement in 5 patients and 3 patients had no follow-up pouchoscopy (Figure 1). There were one patient that had one recurrence after initial successful treatment and another that had two. They were both treated again with FMT successfully. One patient experienced nausea and mild abdominal pain after the treatment, presumably due to ileus. He was hospitalized and treated conservatively. A total of two patients had pouch failure. One was due to bowel obstruction, while the other due to ischemic pouchitis. Conclusions: Our findings suggest that FMT appeared to be a potent alternative for treating recurrent CDI in patients with ileal pouches. Though FMT was effective in the eradication of CDI, the improvement observed on endoscopy was minimal. Table 1. Patient's Clinical Characteristics (N=10).
Su1743 Characterization of Fecal Microbiota in Response to Heterologous Versus Autologous (Placebo) Fecal Microbial Transplantation: Results From a DualCenter, Randomized, Placebo-Controlled Trial Christopher Staley, Colleen R. Kelly, Lawrence J. Brandt, Alexander Khoruts, Michael J. Sadowsky Background: Fecal microbial transplantation (FMT) has been increasingly used as an alternative treatment for recurrent Clostridium difficile infections (RDCI), with a success rate averaging 90%. This treatment has recently been validated in a dual-center, blinded, randomized, placebo-controlled clinical trial, testing the efficacy and safety of healthy donor stool (heterologous FMT) compared with autologous stool, (placebo FMT). Overall, heterologous stool was significantly more effective than autologous (placebo) stool (91% vs 63%), but of interest was the notably higher success rate for autologous FMT observed at the Bronx, NY site (90%) compared with the Providence, RI site (43%). Objective: The fecal microbial compositions of heterologous and autologous FMT recipients were characterized to assess any differences in community composition resulting from fecal material used, pre-FMT treatments, or patient demographic factors. Methods: The fecal microbiome of patients was characterized using Illumina next-generation sequencing of the V5+V6 hypervariable regions of the 16S rRNA gene. Patient samples were collected at baseline, at least 5 days prior to FMT, and again at 2 and 8 weeks post-FMT. Results: Both pre-FMT patient and donor bacterial communities differed significantly at the 2 institutions (P < 0.001). Bacterial communities prior to FMT were dysbiotic and characterized at the phylum level by drastic reduction or disappearance of Bacteroidetes and expansion of the Proteobacteria and/or Verrucomicrobia. Abundances of Verrucomicrobia and Bacteroidetes in pre-FMT samples were notably greater at the Bronx site, where greater autologous FMT success was observed. Heterologous FMT restored the bacterial community alpha diversity to resemble that of healthy donors, while the bacterial community composition in fecal samples of patients also shifted to one similar to donor samples. Community composition after autologous FMT was significantly different than that resulting from heterologous FMT at both sites (P £0.025), and the abundance of Bacteroidetes did not recover to the same extent as was observed in patients who received heterologous donor material. Conclusions: These data reveal that autologous FMT results in different community composition than heterologous FMT, with the latter providing faster and more marked resolution of dysbiotic microbial signatures. Site-specific parameters including antiC. difficile treatment regimens as well as other parameters (e.g. numbers of recurrences and use of proton pump inhibitors) may be driving differences in community composition between centers and subsequent treatment success. Taken together, these results suggest that pre-FMT alteration in the microbiota by anti- C. difficile treatment, for example, may influence the necessity for or success of FMT therapy.
Su1744 Lyophilized Fecal Microbiota Transplantation and Cryoprotectants for Viable Bacteria Preservation Scott W. Mitchell, Anthony Jaworski, Michelle Bull, Connie Wong, Vanessa Furnari, Belinda Chapman, Thomas Borody Introduction: The use of cryoprotectants in foodstuffs, medications and biologic materials has considerable history. However, there is minimal data on the use of cryoprotectants to preserve viable bacteria within lyophilized Fecal Microbiota Transplantation (FMT) product. FMT has received considerable attention for its superior treatment of recurrent Clostridium difficile infection (CDI) compared with antibiotics1. Lyophilized fecal material can be stored for much longer than liquefied stool. The ability to store FMT material for longer relieves the time pressure associated with completion of donor pathology screening to ensure FMT safety. Success of FMT using lyophilized material will depend on preservation of bacterial viability. The choice of cryoprotectant may be important in the optimisation of bacterial viability in lyophilized FMT product. Aim: To measure the total live cell count and compare proportional viability of microorganisms in reconstituted lyophilized FMT following lyophilization using a range of cryoprotectants. Methods: FMT was prepared at a ratio of 1:4 donor stool to saline diluent containing various cryoprotectants. Following homogenization nonbacterial component removal and product concentration were achieved with 250µm filter bags and a refrigerated centrifuge. The resulting slurry was then lyophilized (final water activity £ 0.3). Total and total viable bacterial counts were determined immediately after lyophilization using flow cytometry and Bacteria Count and Live/Dead BacLight ™ kits (Invitrogen). All cryoprotectant formulations were tested at least in triplicate (n = 3 - 6). Total live cell counts were normalised among experiments by expressing the results as the proportion (%) of total live cells. One way analysis of variance was used to compare total live cell proportions among the cryoprotectant formulations. Results: Total live cell proportions immediately following lyophilization ranged from 28.5 - 39.0% (see Table).
AGA Abstracts
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