Based on the known perturbations in microbiome composition and the efficacy of fecal microbial transplantation, microbiome therapies are promising new options for ulcerative colitis (UC). Bacterial consortia as therapeutics have not yet demonstrated effectiveness in UC. An oral microbiome therapeutic is an attractive option for treating mild to moderate UC as monotherapy and potentially in combination with other therapies across the spectrum of disease severity. While probiotics such as Escherichia coli Nissle 1917 (EcN) have marginal efficacy in treating acute flares, they have shown similar efficacy to 5-ASA therapy in remission maintenance. We hypothesized that probiotic and live biotherapeutic product (LBP) efficacy is limited because of failure to colonize the inflamed colon. This failure may be due to vulnerability to oxidative stress. To overcome this, we genetically engineered EcN to enhance colonization. We inserted the ttr operon from Salmonella enterica, which enables tetrathionate (TTR), a gut inflammation byproduct, to be an energy source, providing EcN with a selective advantage during inflammation. EcN::ttr is designed to replicate in response to the degree of underlying inflammation, based on TTR availability, providing a finely tuned therapeutic effect titrated to induce and maintain remission. In both DSS and Muc2-/- murine colitis models, EcN::ttr was significantly more effective in ameliorating colitis than both the unmodified parental strain and 5-ASA therapy. For clinical trials of EcN::ttr, we developed a proprietary oral microencapsulation formulation that optimizes colonic delivery to further enhance colonization. In a 7-day DSS porcine model of colitis, a single dose of orally administered EcN::ttr markedly reduced mortality, with untreated pigs having 20% mortality and severe illness, while treated animals resembled healthy controls clinically and histologically. EcN::ttr treatment preserved gut mucosal integrity and was well tolerated. Two dosing regimens were effective, with evidence of a dose-response for some parameters. This efficacy was achieved with modest doses that can be manufactured efficiently at low cost, providing compelling rationale to advance EcN::ttr as a next-generation microbiome therapeutic into human trials. EcN::ttr may be particularly effective in the maintenance of remission, where the parent EcN strain has previously shown some efficacy albeit in high dosing regimens. A phase 1 trial in 2026 will establish safety and tolerability, followed by an efficacy study initially focused on maintenance of remission and flare prevention in UC. We have shown that the addition of ttr enhances the efficacy of EcN’s anti-inflammatory effects resulting from successful colonization. EcN::ttr may be a novel therapy for UC due to its enhanced ability to successfully colonize the inflamed gut.

Background and Aims: The Mediterranean diet pattern (MDP) is associated with health-associated gut microbes and metabolites. However, the impact of the MDP on the fecal metabolome in ulcerative colitis (UC) remains unclear. We characterized the fecal metabolome of patients with UC with high adherence to the MDP compared to the Canadian habitual diet (CHD). Furthermore, we explored potential differences in the fecal metabolome between dietary responders and nonresponders to the MDP. Methods: Utilizing untargeted metabolomics on a subset of fecal samples obtained from a randomized controlled trial, adult patients with quiescent UC underwent a 12-week intervention following either the MDP (n = 8) or CHD (n = 8). Liquid chromatography-tandem mass spectrometry was employed to profile endogenous fecal metabolites, while 16S amplicon sequencing was utilized to profile the fecal microbiota. Results: A total of 701 human metabolites were detected, with 35 exhibiting significant differential expression between the MDP and CHD groups. Noteworthy, folate biosynthesis, sphingolipid biosynthesis, and steroid biosynthesis were identified as major pathways affected. Moreover, microbial analysis showed that individuals with increased levels of the class Bacteroidia (Bacteroides vulgatus [B. vulgatus], B. uniformis, and B. acidifaciens) in their stool at baseline were more likely to respond to the MDP. Conclusion: High adherence to an MDP is associated with beneficial metabolite changes associated with reducing inflammation in UC. In addition, fiber-degrading microbes abundant before dietary intervention played a role in the responsiveness to the MDP. This work lays the groundwork for developing a metabolic signature associated with the MDP to develop personalized nutrition strategies for UC prevention and treatment. ClinicalTrials.gov Number: NCT03053713.

The gut microbiota of mammals possess distinctive metabolic pathways with untapped therapeutic potential. Using molecular insights into dietary fiber metabolism by the human gut microbiota, we designed a targeted drug delivery system, called GlycoCaging, that is based on bespoke glycoconjugates of a complex plant oligosaccharide. GlycoCaging of exemplar anti-inflammatory drugs enabled release of active molecules triggered by specific glycosidases of autochthonous gut bacteria. GlycoCaging ensured that drug efficacy was potentiated, and off-target effects were eliminated in murine models of inflammatory bowel disease. Biochemical and metagenomic analyses of gut microbiota of individual humans confirmed the broad applicability of this strategy.

Background: Dietary fatty acids (FA) are crucial to the pathophysiology of inflammatory bowel disease (IBD), influencing systemic and gut inflammatory responses. Dietary FA intake influences the fatty acid profiles of vital cell membranes, which might be a source of inflammatory mediators. Despite their significance, research on dietary FA subtypes and their effects on inflammation and oxidative stress in IBD is limited. Objective: We investigated the association between dietary FA intake, the erythrocyte membrane FA composition (EMFA), and inflammation and oxidative stress markers in patients with mild–moderate luminal Crohn’s Disease (CD) participating in the CD Therapeutic Dietary Intervention (CD-TDI). Design: A cross-sectional analysis was performed on 24 participants (13 CD-TDI, 11 habitual diet controls) from a 13-week randomized controlled trial assessing the efficacy of CD-TDI in inducing clinical and biomarker remission in CD. Methods: EMFA was analyzed using direct-injection gas chromatography, and dietary FA intake was assessed using the ASA 24-h Dietary Assessment Tool ® . Results: The CD-TDI group showed a significant increase in dietary n-3 polyunsaturated fatty acids (PUFA) at Week 13 ( p = 0.04) compared to no changes in the control group. Participants on the CD-TDI also demonstrated a significant reduction in total fat, saturated fat, and arachidonic acid (AA) ( p < 0.01). EMFA analysis revealed lower percentages of AA ( p = 0.03) in the CD-TDI group. Positive correlations were observed between C-reactive protein, fecal calprotectin, and dietary stearic acid ( p < 0.05). Inverse correlations were found between malondialdehyde (MDA) and the Mediterranean Diet Score ( r = −0.67) as well as MDA and the intake of whole fruit, legumes, and nuts/seeds ( r > −0.50). Conclusion: The CD-TDI significantly increased dietary n-3 PUFA intake, reduced pro-inflammatory n-6 PUFA (AA), and improved markers of oxidative stress, supporting its potential in CD management. The cell membrane fatty acid profile might be a therapeutic target in CD. Trial registration: NCT04596566.

Indian immigration to westernized countries has recently surged, increasing their risk of Inflammatory Bowel Disease (IBD) post-migration. While crucial for understanding IBD risk, the gut microbiome remains understudied in Indians. This cross-sectional study examines the impact of westernization on the gut microbiomes of Indians residing in India, Indo-Immigrants, and Indo-Canadians compared to Euro-Canadian and Euro-Immigrant controls. Stool samples for 16S rRNA and shotgun sequencing assessed microbial taxa and functional profiles, alongside dietary and demographic data to evaluate lifestyle patterns. Indians and Indo-Immigrants had distinct microbiotas from controls, with high abundances of Prevotella spp. and CAZymes reflecting their high complex carbohydrate diet. Indo-Canadians exhibited a transitional microbiome towards westernization, mirroring increasing dietary acculturation. Considering 44% of Canadians are first- and second-generation immigrants and the global adoption of westernized practices, future research should investigate the health implications of such microbiome transitions in immigrant populations and newly industrialized nations.

Glyphosate, a widely used herbicide in North America, has become prevalent in the food supply due to preharvest applications, raising concerns about potential health impacts. This study investigated the effects of prenatal glyphosate exposure on the gut bacteriome, colitis, metabolic health, and behavior across generations in mice. In healthy mice, glyphosate decreased goblet cell numbers and mucin-2 expression in the colon and dysregulated cytokines in F1 and F2 progeny at levels below the EPA acceptable daily limit. Glyphosate also disrupted metabolism, including impaired glucose tolerance, increased insulin resistance and reduced serum GLP-1 levels. Moreover, prenatal glyphosate exposure induced behavioral deficits, including reduced locomotor activity and impaired working memory, which are associated with alterations in the gut microbiome composition and key gut-brain axis mediators. These data underscore the potential risks associated with glyphosate exposure, highlighting the need for further research and regulatory consideration. ### Competing Interest Statement The authors have declared no competing interest.

Background Indian immigration to westernized countries has surged in recent years, with this demographic facing a higher risk of Inflammatory Bowel Disease (IBD) after migration. Studying the gut microbiome is crucial to understanding the factors contributing to IBD, yet Indians remain underrepresented in microbiome research, Objective This study investigates the gut microbiome of Indians residing in India, Indian immigrants in Canada, and Canadians of Indian ancestry to understand the impact of westernization on their gut. Given the higher risk of IBD among Indo-Canadians, these results provide insight into how westernization of the gut microbiome may influence incidence of IBD. Stool samples were collected from healthy Indians in India, Indian Immigrants in Canada, Indo-Canadians, plus Canadian-borne individuals with European descent and Euro-Immigrants as non-Indian/westernized controls ( N = 174). 16S rRNA gene and shotgun sequencing compared microbial taxa and functional profiles across groups. Dietary and demographic data were collected to assess lifestyle patterns. Results Indians and Indo-Immigrants had distinct gut microbiotas from westernized groups, with high abundances of Prevotella spp. and a CAZyme profile reflecting their traditional high complex carbohydrate diet. The Indian microbiome also showed characteristics of high bacterial cell turnover, pathogenic potential, and stress tolerance, indicating their gut may be better primed to handle stressors on the gut. Indo-Canadians displayed a transitional microbiome towards westernization, which followed a pattern of increasing dietary acculturation. Conclusions The shift from a non-westernized to westernized microbiome in Indo-Canadians highlights the significant influence that westernization and dietary acculturation has on the gut microbiome. Given that 44% of the Canadian population is made up of first- and second-generation immigrants and westernized practices are being adopted worldwide, future research should investigate the health implications that such microbiome transitions have on immigrant populations and newly industrialized nations.

Abstract Background Inflammatory bowel disease (IBD) is characterized by chronic inflammation in the gastrointestinal tract. Despite the alarming global increase in IBD burden with 1 in 121 Canadians affected, no cure exists. Systemic administration of anti-inflammatory drugs like corticosteroids is a front-line therapy for inducing remission. However, these drugs are limited in their application by deleterious off-target side effects. To target the drug to sites of inflammation and reduce systemic uptake, we developed a microbiome-cleavable, glycoconjugate drug delivery system, termed GlycoCage. We have demonstrated that GlycoCaged dexamethasone (Dex, a potent steroid), is effective at reducing inflammation at 10-fold lower doses than its uncaged counterpart in the SHIP-deficient (SHIP-/-) mouse model of CD-like intestinal inflammation. We hypothesize that GlycoCaged technology can expand the therapeutic window of IBD drugs by lowering effective doses and eliminating side effects. Aims: 1) Quantify Dex in systemic circulation. 2) Assess status of inflammation outside of the gut in SHIP-/- mice treated with caged Dex. 3) Measure decaging activity in people with IBD. Methods To compare systemic distribution of Dex to caged Dex, we orally gavaged SHIP+/+ and SHIP-/- mice with 3 mg/kg of Dex or the molar equivalent of caged Dex and collected serum for LC-MS/MS analysis at multiple timepoints post oral gavage. In addition to CD-like ileitis, SHIP-/- mice spontaneously develop enlarged mesenteric lymph nodes (MLNs) and inflamed lungs. After daily treatment of Dex or caged Dex for 2 weeks, MLNs and lungs were collected and analyzed for size and histopathology. We also obtained stool samples from healthy control participants and people with IBD to investigate the prevalence of prodrug-cleaving activity by direct enzymatic activity assay using fluorogenic resorufin as a prodrug proxy. Data from people with IBD were stratified to fecal calprotectin. Results Unlike free dexamethasone, caged dexamethasone did not enter systemic circulation or impact inflammation outside of the gut in SHIP-/- mice. All people have decaging activity that is not affected by IBD or the individual’s disease activity. Conclusions Our results highlight a novel glycoconjugated prodrug strategy to improve and broaden treatment options for people with IBD. In the future, we will evaluate GlycoCaged Dex in a mouse model of colitis and test the efficacy of other GlycoCaged drugs. Funding Agencies CCCGlycoNet

The O-glycoprotein Mucin-2 (MUC2) forms the protective colon mucus layer. While animal models have demonstrated the importance of Muc2, few studies have explored human MUC2 in similar depth. Recent studies have revealed that secreted MUC2 is bound to human feces. We hypothesized human fecal MUC2 (HF-MUC2) was accessible for purification and downstream structural and functional characterization. We tested this via histologic and quantitative imaging on human fecal sections; extraction from feces for proteomic and O-glycomic characterization; and functional studies via growth and metabolic assays in vitro. Quantitative imaging of solid fecal sections showed a continuous mucus layer of varying thickness along human fecal sections with barrier functions intact. Lectin profiling showed HF-MUC2 bound several lectins but was weak to absent for Ulex europaeus 1 (α1,2 fucose-binding) and Sambucus nigra agglutinin (α2,6 sialic acid-binding), and did not have obvious b1/b2 barrier layers. HF-MUC2 separated by electrophoresis showed high molecular weight glycoprotein bands (∼1-2 MDa). Proteomics and Western analysis confirmed the enrichment of MUC2 and potential MUC2-associated proteins in HF-MUC2 extracts. MUC2 O-glycomics revealed diverse fucosylation, moderate sialylation, and little sulfation vs. porcine colonic MUC2 and murine fecal Muc2. O-glycans were functional and supported the growth of Bacteroides thetaiotaomicron (B. theta) and short-chain fatty acid (SCFA) production in vitro. MUC2 could be similarly analyzed from inflammatory bowel disease stools, which displayed an altered glycomic profile and differential growth and SCFA production by B. theta vs. healthy samples. These studies describe a new non-invasive platform for human MUC2 characterization in health and disease.

Fasting diets (FDs) have drawn great attention concerning their contribution to health and disease over the last decade. Despite considerable interest in FDs, the effect of fasting diets on eating behaviors, sleep, and mood-essential components of diet satisfaction and mental health- has not been addressed comprehensively. Understanding the critical role that fasting plays in these elements will open up potential treatment avenues that have not yet been explored. The aim of the present paper was to conduct a comprehensive critical review exploring the effects of fasting on eating behaviors, sleep, and mood. There is currently a lack of clarity regarding which fasting option yields the most advantageous effects, and there is also a scarcity of consistent trials that assess the effects of FDs in a comparable manner. Similarly, the effects and/or treatment options for utilizing FDs to modify eating and sleep behaviors and enhance mood are still poorly understood. Further researches aiming at understanding the impacts of various fasting regimes, providing new insights into the gut-brain axis and offering new treatment avenues for those with resistant anxiety and depression, are warranted. Alteration of eating behaviors can have lasting effects on various physiological parameters. The use of fasting cures can underpin ancient knowledge with scientific evidence to form a new approach to the prevention and treatment of problems associated with co-morbidities or challenges pertaining to eating behaviors. Therefore, a thorough examination of the various fasting regimens and how they impact disease patterns is also warranted.

Abstract Background Young Indian immigrants and Indo-Canadians face a significantly higher risk of inflammatory bowel disease (IBD) in westernized countries. While the etiology of IBD remains unclear, a gut microbiome that is no longer symbiotic with its host is a key player. However, Indians are underrepresented in microbiome research, therefore we cannot accurately assess the role of their gut microbiome in IBD. To understand why Indians are at a greater risk for IBD in Canada, we must first characterize their gut microbiome. Aims This study explores how immigration to Canada impacts the gut microbiome of Indian populations, potentially elevating their risk to IBD. Methods Stool samples from healthy volunteers (ages 17-53) were collected from Indians in India, Indo-Immigrants, Indo-Canadians, Euro-Canadians and Euro-Immigrants. DNA was extracted for 16S sequencing on the Illumina MiSeq platform and shotgun sequencing on the Illumina NovaSeq platform. Dietary data was processed in ESHA. Results Weighted Unifrac revealed distinct clusters of Indian and Indo-Immigrant samples from the westernized cohorts. The most pronounced difference was between Indian and Euro-Canadians with a pseudo-F value of 49.18 (q = 0.00125). Indian versus Indo-Immigrants resulted in a pseudo-F value of 7.707 (q = 0.00125), and Indian versus Indo-Canadian had a pseudo-F value of 15.95 (q = 0.00125). BugBase estimated a significantly higher stress tolerance in the Indian gut, with Proteobacteria driving this phenotypic prediction. Compared to Euro-Canadians, a significantly higher pathogenic potential was predicted in Indians (pFDR= 1.04e-11) and Indo-Immigrants (pFDR= 1.41e-04), driven mainly from Proteobacteria and Bacteroidetes. Multiple comparisons with LDA revealed Prevotella was over 5 times more abundant in Indians residing in India. The gut microbiome in Indians were also enriched with taxa associated with a plant-based diet. Dietary data showed that 59% of Indians do not consume meat, and they also had the lowest consumption (12% caloric intake) of group 4 ultra-processed food (UPF), whereas the highest consumption of UPF was in Indo-Canadians (61% caloric intake) (pbonf ampersand:003C 0.0001). A frequent trend was observed in Indians living in Canada, which was a reduction in key nutrients including soluble fiber, zinc, iron, and folate, some of which are commonly found in plant-based foods. Conclusions Overall findings reveal that a loss of Prevotella abundance was observed in Indo-Immigrants and Indo-Canadians, which was associated with a reduction in key nutrients that are common to a plant-based diet, indicating a transition away from their traditional diet. Their dietary changes may be a driver to the displacement of Prevotella and other taxa commonly found in the Indian gut microbiome. Funding Agencies NSERC, Killam

The intricate balance between the gut microbiome and host health inspires innovations in drug development. Commensal bacteria provide a multi-targeted approach ideal for treating complex medical conditions, like inflammatory bowel disease (IBD). These bacteria are self-replicating factories with broad targets that promote balanced intestinal inflammation, mucosal barrier function, and eubiosis. Yet, the lack of superiority to gold-standard treatments and their clinical inconsistency makes most probiotics unreliable for disease treatments. Intestinal inflammation, a driving factor in many diseases, often overwhelms commensal bacteria, which lack the stress-resistance mechanisms necessary to withstand host immune defenses. To address this, we introduced a persistence platform BioPersist™ into E. coli Nissle 1917. We hypothesized that a bio-engineered probiotic, or genetically engineered microbial medicine (GEMM™), designed to persist during inflammation would enhance probiotic bioavailability during colitis, leading to sustained therapeutic outcomes. We evaluated BioPersist in multiple translational colitis models such as in mice and pigs. BioPersist delayed the onset and reduced the severity of both chronic and acute colitis, proving more effective than 5-aminosalicylate. BioPersist thrived during inflammation promoting tolerogenic immune responses that limited infiltrating leukocyte activity and decreased TNF-α from resident myeloid cells in the mesentery. The persistence feature of BioPersist allowed the probiotic to overcome the damaging inflammatory response, eliciting mucosal healing evident by the increase in microbially-derived butyric acid. Based on these preclinical results, BioPersist may be a novel therapeutic option for both human and veterinary applications that sustains efficacy during colitis. ### Competing Interest Statement DLG is a co-founder and Chief Scientific Officer of Melius MicroBiomics Inc, a start-up company that has licenced the UBC technology under the following patents. PCT/CA2018/050188: D.L Gibson, A. Godovanny, S.K. Gill, Designer probiotics engineered to efficiently colonize the gut for effective inflammatory bowel disease (IBD) therapy. Tech Transfer 2023 with Melius MicroBiomics Inc; and 63/704,043 (patent pending). DLG and AG have founding shares in Melius MicroBiomics Inc. AG, JAB and MKA are Melius MicroBiomics Inc employees. ME, MKA, AG, JAB, DLG have Melius MicroBiomics Inc. options. LMS, AV, SKG, NH, JY, RI, HM, JKJ and CG declare that they have no competing interests.

BACKGROUND AND AIMS:Dietary patterns are important in managing ulcerative colitis [UC], given their influence on gut microbiome-host symbiosis and inflammation. We investigated whether the Mediterranean Diet Pattern [MDP] vs the Canadian Habitual Diet Pattern [CHD] would affect disease activity, inflammation, and the gut microbiome in patients with quiescent UC. METHODS:We performed a prospective, randomised, controlled trial in adults [65% female; median age 47 years] with quiescent UC in an outpatient setting from 2017 to 2021. Participants were randomised to an MDP [n = 15] or CHD [n = 13] for 12 weeks. Disease activity [Simple Clinical Colitis Activity Index] and faecal calprotectin [FC] were measured at baseline and week 12. Stool samples were analysed by 16S rRNA gene amplicon sequencing. RESULTS:The diet was well tolerated by the MDP group. At week 12, 75% [9/12] of participants in the CHD had an FC >100 μg/g, vs 20% [3/15] of participants in the MDP group. The MDP group had higher levels of total faecal short chain fatty acids [SCFAs] [p = 0.01], acetic acid [p = 0.03], and butyric acid [p = 0.03] compared with the CHD. Furthermore, the MDP induced alterations in microbial species associated with a protective role in colitis [Alistipes finegoldii and Flavonifractor plautii], as well as the production of SCFAs [Ruminococcus bromii]. CONCLUSIONS:An MDP induces gut microbiome alterations associated with the maintenance of clinical remission and reduced FC in patients with quiescent UC. The data support that the MDP is a sustainable diet pattern that could be recommended as a maintenance diet and adjunctive therapy for UC patients in clinical remission. ClinicalTrials.gov no: NCT0305371.

Abstract Background Live biotherapeutic products (LBPs) offer a more rationalized and multitargeted approach to treating gastrointestinal diseases. BioColoniz and BioPersist are two LBPs derived from the parental strains L. reuteri and E. coli Nissle 1917 (EcN), respectively. The parental strains are known to offer some benefit in preventing relapses in IBD patients however the results are heterogeneous. To overcome this, the parental strains were approached as LBPs by introducing traits to thrive under the inflammatory conditions of the colon. Therefore, our aim is to characterize the role of these LBPs in IBD. Purpose To evaluate the therapeutic effect of the LBPs BioColoniz and BioPersist in an acute model of colitis. Method Female C57Bl/6 mice were treated with BioColoniz or BioPersist via oral gavage for three consecutive days prior to DSS challenge. Then mice were exposed to 3.5% DSS via drinking water for seven days. As controls, we also included mice treated with vehicle or the parental strains L. reuteri or EcN. In order to compare the effect of LBPs in the onset of acute colitis to current maintenance therapies for UC, we also exposed another group of mice to DSS and simultaneously administer 5-ASA. Mice were monitored daily for signs of disease and at the end of the experiment, colon tissue was collected for histopathological and molecular analysis. Result(s) The administration of BioColoniz and BioPersist delayed and decreased the colitic phenotype of mice exposed to DSS. Differences in signs of disease, such as diarrhea and weight loss, were evident by day 4 for vehicle or 5-ASA groups, whereas mice in the LBPs groups were still gaining weight. When analyzing the histopathological changes, mice in the LBPs groups presented lower scores when compared to the vehicle and 5-ASA groups. Specifically, mice treated early with BioColoniz or BioPersist presented a more preserved mucosal architecture with visible crypts. Although 5-ASA-treated mice still had vestiges of crypts, the damage in the mucosal architecture was more severe, similar to the observed in mice treated with the parental strains EcN and L. reuteri. We also looked at the expression of proinflammatory cytokines, finding an increase in TNFα, IFNγ, and IL-17a in mice treated with 5-ASA but not in mice treated with LBPs. However, the expression of protective factors such as mucin Muc2 or the antimicrobial peptide Reg3γ was similarly high in 5-ASA and LBP-treated mice when compared to vehicle or parental strains groups, suggesting some therapeutic commonalities between 5-ASA and our LBPs. Conclusion(s) The early administration of the LBPs BioColoniz and BioPersist protect mice from severe acute colitis, being more protective than 5-ASA. Since some differences and similarities were observed between the LBPs- and 5-ASA-treated mice, such as crypt preservation versus increased expression of some protective factors, the next step will aim to identify which mechanisms are specifically triggered by the LBPs. Disclosure of Interest None Declared

In January 2022, a group of experts came together to discuss current perspectives and future directions in nutritional immunology as part of a symposium organized by the Canadian Nutrition Society. Objectives included (1) creating an understanding of the complex interplay between diet and the immune system from infants through to older adults, (2) illustrating the role of micronutrients that are vital to the immune system, (3) learning about current research comparing the impact of various dietary patterns and novel approaches to reduce inflammation, autoimmune conditions, allergies, and infections, and (4) discussing select dietary recommendations aimed at improving disease-specific immune function. The aims of this review are to summarize the symposium and to identify key areas of research that require additional exploration to better understand the dynamic relationship between nutrition and immune function.

Abstract Background During the 1900s when industrialization was on the rise in western countries, inflammatory bowel disease (IBD) began to present itself and continually increase over the decades. Now, newly industrialized countries such as India are following this same pattern, and with a population reaching over one billion, India is projected to have one of the highest IBD prevalence worldwide. Furthermore, pediatric diagnoses of IBD are more frequently reported in India and in Indian children living in Canada, suggesting this disease may present differently in those of Indian descent. While the etiology of IBD remains unclear, a gut microbiome that is no longer symbiotic with its host is a key player. However, Indians are one of the least represented in microbiome research, therefore we cannot accurately assess the role of their gut microbiome in IBD. To effectively understand the nature of IBD in Indians, we must first define their gut microbiome. Purpose Our study characterizes the microbiome of Indians living in India to explore how it differs from Canadians of European descent. Method Stool samples from healthy volunteers (ages 18-55) were collected from Indians in India and Euro-Canadians in Kelowna, BC. Microbial DNA was extracted for 16S sequencing on the Illumina MiSeq platform and QIIME2 was used for microbiome analysis. Result(s) We will discuss the similarities and differences comparing the gut microbiome of Indians to Euro-Canadians, further highlighting the need for more microbiome research of this demographic. Conclusion(s) Our research aims to increase representation of Indians in microbiome research in the hopes of improving our knowledge of the predispositions to IBD, which will aid in the information required to develop effective preventive measures. With elevated risk for IBD in Indians residing in Canada, future studies should also aim to analyze if the gut microbiome in Indians change as they migrate and adopt the westernized lifestyle. Disclosure of Interest None Declared

Abstract Background Canada is one of the most prolific users of the herbicide glyphosate (tradename Roundup®), with over 25 million kilograms purchased annually. Glyphosate is commonly applied pre-harvest as a desiccant, leading to higher residues in many foods consumed within Canada, including wheat, cereals, legumes, and soy products. Glyphosate prevents the synthesis of aromatic amino acids by inhibiting the Shikimate pathway, which, besides killing weeds, can also inhibit bacterial growth and promote bacterial dysbiosis. Dysbiosis has been associated with multiple disease states, including intestinal inflammation and metabolic dysfunction, which both include anxiety, depression, and cognitive dysfunction as co-morbidities. Purpose My goal is to clarify if there are harmful glyphosate effects on the gut bacteriome, resulting in altered metabolic health, damaging intestinal inflammation, and behaviour changes through the gut-brain axis. We explored two environmentally relevant doses throughout our experiments: (1) the acceptable daily intake currently set by the Environmental Protection Agency (EPA) (1.75mg/kg body weight/day); (2) the North American dose calculated with a registered dietician based on literature values of glyphosate within items that make up a typical Canadian diet (0.01mg/kg body weight/day.) Method Breeding pairs consisting of Muc2-/- (colitis susceptible) and Muc2+/- (littermate control) mice were exposed to glyphosate during pregnancy. A subset of each generation underwent behavioural testing to characterize anxiety, memory, and locomotor activity. Animals also underwent metabolic testing, including oral glucose and insulin tolerance tests. Only the parental generation of animals received glyphosate. In total, three generations of animals were raised. Result(s) Our results show that healthy mice whose great-grandparents were exposed to glyphosate at levels currently deemed safe by the EPA exhibited decreased locomotor activity. Furthermore, mice whose parents were exposed to either North American or EPA glyphosate levels exhibit symptoms of metabolic dysfunction in healthy and colitis-susceptible mice, including higher fasting blood glucose, an inability to efficiently clear glucose and impaired insulin response. Additionally, Muc2-/- mice whose parents were exposed to levels of glyphosate found within the North American diet during pregnancy showed significant impairments in working memory. Conclusion(s) This study is the first to highlight the transgenerational effects of glyphosate at levels previously deemed safe for human exposure on locomotor activity, working memory and metabolism. Additionally, this study highlights how environmental toxins within our food system, such as glyphosate, may play a vital role in the etiology of many diseases in healthy and colitis-susceptible populations by promoting a sedentary lifestyle, metabolic dysfunction, and behavioural deficits. Please acknowledge all funding agencies by checking the applicable boxes below CAG, CCC, Other Please indicate your source of funding; NSERC Disclosure of Interest None Declared

Abstract Background Inflammatory Bowel Diseases (IBD) are increasing globally with westernized countries experiencing a dramatic increase in cases of ulcerative colitis and Crohn’s disease. The corresponding rise in the comorbidities of anxiety and depression are linked to inflammation and microbiome dysbiosis; impacting the gut-brain axis (GBA). Another facet of the GBA are the tryptophan metabolites serotonin (5HT) and kynurenine (Kyn). 5HT and Kyn are metabolites implicated in anxious behaviours and memory deficiencies with dysbiosis influencing metabolite production. In inflammatory conditions, upregulation of Kyn impacts brain homeostasis as Kyn has both positive and negative brain metabolites. As disease outcomes are tied to the severity of depression/anxiety, characterization of the behavioural and tryptophan metabolite profile of an IBD model can allow us to develop better IBD therapies. Purpose To characterize the anxiety and memory phenotype with corresponding assessment of tryptophan metabolites 5HT and Kyn across tissues of interest in an spontaneous colitis model. Method Mucin 2 knockout mice (Muc2-/-) lack the protective mucin 2 layer in the distal colon, allowing for direct contact between the gut contents and the gut epithelial layer. Over time this contact results in the development of spontaneous colitis. Muc2-/- n=12 per sex were compared to C57Bl/6 mice for behaviour and tryptophan metabolites across brain, colon and serum. Open Field Test and Light/Dark tests were used to assess anxiety levels, Novel Object and Radial Arm maze were performed to assess memory deficits. Mice underwent one anxiety and one memory test and were euthanized following their last maze. Enzyme-linked immunosorbent assay (ELISA) assessment of 5HT and Kyn were performed on brain, colon and serum samples. T-tests were performed and graphs created using PRISM (v9.4.1). Result(s) Muc2-/- mice had decreased anxiety (p=0.0113) compared to C57Bl/6 wild type mice in the light/dark maze. Muc2-/- displayed no significant differences in novel object preferences or the number of memory errors in novel object recognition test and radial arm maze respectively. Serum kynurenine was found to be significantly elevated (p=0.0444) in Muc2-/- mice. The sex factor has to be mentioned here in the results not in the conclusions. Conclusion(s) IBD is increasing globally and problems with mental health is often comorbid. Within our model we were able to show that behavioural and biochemical differences do exist between our Muc2-/-and wild-type mice with depressed anxiety responses and heightened serum kynurenine, however the implications of these findings require further investigation. Understanding how inflammation is driving behavioural alterations can provide pathways to ameliorate the behavioural comorbidities in IBD patients suffering from anxiety of memory deficits. Disclosure of Interest None Declared