Introduction

It is known that disorders of the intestinal microbiota in patients with ulcerativ colitis (UC) lead to increased intestinal permeability, impaired production and intestinal absorption of short-chain fatty acids (SCFA). The ratio of fecal SCFA is an important indicator of the integrity of the intestinal microbial community. The study of the concentrations of SCFA is a very attractive approach to broadening the understanding of the pathogenesis of UC.

Aims & Methods

To study the content of SCFA in the coprofiltrate of patients depending on the degree of severity of the course of UC.
We included in this study 86 patients with UC with a mean age of 43.2 ± 1.8 years. They were divided according to the Mayo index: 7 milds, 64 moderate and 15 severe. The control group consisted of 7 relatively healthy people. The diagnose UC were established according to generally accepted diagnostic standards in gastroenterology. Determination of SCFA was carried out by the method of chromatographic research. Was determined anaerobic index (AI) - the ratio of the sum of propionic and butyric acids concentrations to the concentration of acetic acid. AI values are used as an indicator of the infrastructure of the microbiocenosis - the ratio of anaerobic and aerobic microorganisms.

Results

The total content of SCFA did not show significant changes in patients with mild and moderate severity of UC (only a moderate tendency to decrease by 1.6 times (p>0.05) and 1.8 times (p>0.05), respectively), while in severe severity there was a statistically significant decrease by 3.9 times (p=0.008) compared to control. Also, in severe UC, this indicator had a pronounced tendency to decrease by 2.4 times (p>0.05) and 2.2 times (p>0.05), respectively, compared with mild and moderate severity, respectively. In mild and moderate severity of the disease, only a weak tendency to reduce the content of acetic acid in the coprofiltrate of patients by 1.4 times (p>0.05) and 1.3 times (p>0.05), respectively, and in severe severity - a significant decrease by 1.7 times (p=0.02), compared to the control. The content of propionic acid in the experimental samples of patients did not change significantly, there was only a moderate/weak tendency to its decrease by 1.6 times (p>0.05), 1.3 times (p>0.05) and 1.2 times (p>0.05), respectively, for the three severity levels of UC compared to the control. A significant decrease in the content of butyric acid in the coprofiltrate of patients with moderate and severe UC severity was found by 2.1 times (p=0.049) and 10.4 times (p=0.001), respectively, compared to the control. It is characteristic that in severe UC, the content of butyric acid was significantly reduced by 5.7 times (p=0.034) and 5 times (p=0.022), respectively, compared with mild and moderate severity of this disease.
IA significantly decreased by 1.5 times (p=0.028) and 1.9 times (p=0.033) in moderate and severe UC severity compared to the control group.
Thus, in severe UC severity, there was a marked decrease in the total content of FFAs mainly due to a significant decrease in butyric acid and a moderate decrease in acetic acid.

Conclusion

The obtained results showed that changes in SCFA concentrations in patients with UC differed significantly from the control group, what can lead to deterioration of the course and prognosis of UC. Estimation of the ratio of SCFA fractions with the calculation of the anaerobic index may be useful for the diagnosis of violation of the quantitative and qualitative composition of the intestinal microbiota in patients with UC.

References

1. Parada Venegas D, De la Fuente MK, Landskron G, et al. Short Chain Fatty Acids (SCFAs)-Mediated Gut Epithelial and Immune Regulation and Its Relevance for Inflammatory Bowel Diseases. Front Immunol. 2019;10:277. Published 2019 Mar 11. doi:10.3389/fimmu.2019.00277
2. Shin Y, Han S, Kwon J, et al. Roles of Short-Chain Fatty Acids in Inflammatory Bowel Disease. Nutrients. 2023;15(20):4466. Published 2023 Oct 21. doi:10.3390/nu15204466
3. Lee S.H., Kwon J.Е., Cho M.L. Immunological pathogenesis of inflammatory bowel disease. Intestinal Research, 2018. Vol. 16, №1. Р.26-42. doi: 10.5217/ir.2018.16.1.26.

Introduction

Inflammatory bowel diseases (IBD) are mainly represented by Crohn's Disease (CD) and Ulcerative Colitis (UC). Among several factors related to these diseases, dysbiosis is considered an immediate environmental factor for the development of IBD. Dysbiosis is related to the imbalance of the intestinal microbiota, which affects the existing symbiotic relationship between the intestinal microbiota and its host that provides a nutrient-rich environment for intestinal bacteria, while the microbiota plays a fundamental role in maintaining the intestinal function of its host. Among these functions, the microbiota has its participation in the digestion of complex polysaccharides present in the host diet, important for the formation of monosaccharides and short-chain fatty acids (SCFA), such as acetate, propionate, and butyrate. These SCFA are important in maintaining the intestinal barrier and intestinal homeostasis, in proliferation, differentiation and modulation of gene expression of intestinal cells, in the production of essential vitamins in the intestinal lumen, in addition to acting in regulation of the mechanisms that provide balance between the intestinal mucosa and its immune system, resulting in the tolerance of harmless bacteria and immune responses against pathogenic bacteria. There are several studies on the intestinal microbiota in patients with IBD, however, there are still few studies on the SCFA profile in these patients. Therefore, the present study aimed to evaluate and compare the SFCA profile in patients with IBD and healthy individuals.

Aims & Methods

94 patients from a public outpatient healthcare service in Brazil and 63 healthy individuals were included in the study. Exclusion criteria: diagnosis of other chronic diseases, use of antibiotics, prebiotics, probiotics or symbiotics in the last 4 weeks. Participants were divided into two groups, IBD group and control group (healthy), fecal samples from all participants were collected for qualitative and quantitative analysis of SCFA, which was performed by gas chromatography technique coupled with mass spectrometry (GC/MS). Statistical analysis: descriptive statistics and association tests. Values of p ≤ 0.05 were considered significant. Data presented in mean (SD).

Results

The SCFA profiles were different in IBD patients compared to healthy individuals, with higher SCFA values in the IBD group when compared to the control group: acetic acid (0.32 ± 0.18 vs 0.12 ± 0.11, with p< 0.0001), propionic acid (0.24 ± 0.10 vs 0.15 ± 0.09, with p< 0.0001), butyric acid (0.19 ± 0.11 vs 0.39 ± 0.25, with p< 0.0001) and sum of SCFA (0.75 ± 0.32 vs 0.39 ± 0.25, with p< 0.0001) concentration.

Conclusion

The SFCA profile in patients with IBD showed a difference when compared to healthy individuals, where the values of SFCA fecal were higher in patients with IBD compared to healthy individuals. Such a difference may occur because the IBD group, due to the inflammatory process characteristic of the disease, may have greater difficulty in absorbing the SCFA produced in the intestine, releasing more SCFA in the feces, since most of the SCFAs produced in the colonic lumen (90-95%) are absorbed by the intestinal mucosa and only 5 to 10% are excreted in the feces. However, to confirm this hypothesis, further studies would be needed to evaluate and compare the SFCA systemic between these groups, through blood test, and thus determine the SFCA profile of these individuals, being able to propose therapies that help in the modulation of the microbiota of these patients and in the development of new treatment targets in the future.

References

1. García‐Villalba R, et al. Journal of separation science. 2012; 35(15): 1906-1913.
2. Kaplan GG. Nature reviews Gastroenterology & hepatology.2015; 12(12): 720-727.
3. Sánchez-Tapia M, Tovar AR, Torres N. Archives of Medical Research. 2019; 50(5): 259–268.
4. Schirmer M, et al. Nature Reviews Microbiology. 2019; 17(8): 497-511.
5. Zhao G, Nyman M, and Jönsson, A J. Biomedical Chromatography. 2006; 20(8): 674-682.

Introduction

Numerous studies show that changes in the composition of the gut microbiota constitute an important factor influencing both the development of ulcerative colitis (UC) and the prognosis of the disease, drug metabolism and effectiveness of treatment. The growing global prevalence of UC necessitates continued advances in diagnostics and treatment. Gut microbiota can be considered as a promising dynamic biomarker for non-invasive assessment of the severity of UC and the effectiveness of treatment.

Aims & Methods

The purpose of the research was to study the taxonomic diversity of the gut microbiota in patients with UC at different stages of the disease. The study included 21 patients (ages from 20 to 41, 13 men and 8 women) in the relapse stage of UC - left-sided UC with mild to moderate activity (Truelov-Witts scale, Mayo index); and 20 patients (ages from 26 to 52, 10 men and 10 women) in the stage of clinical endoscopic remission. The study of the coprofiltrate microbiota was carried out by NGS sequencing of the structure of the bacterial 16S ribosomal RNA gene using the Illumina MiSeq platform. Statistical analysis was preceded by testing of the distribution for normality using the Kolmogorov-Smirnov (with Liljefors significance correction) and Shapiro-Wilk tests. Hypotheses about the distribution corresponding to normal were rejected, and therefore the calculation of descriptive statistics is presented by median and quartiles. Group comparisons were made using the nonparametric Mann-Whitney test.

Results

Statistically significant differences between the relapse and remission groups were identified for 15 genera of microorganisms. In the relapse group, higher levels of Enterococcus, Streptococcus, Granulicatella, Pelomonas, TM7x, Actinomyces, Gemella, Corynebacterium, Bradyrhizobium, and lower levels of NK4A214_group, Pseudoscardovia, Family_XIII_UCG-001, Anaerofilum, Candidatus_Soleaferrea, DNF00809 were found (Table 1).

Conclusion

Data obtained demonstrates taxonomic differences at the genus level in patients in the relapse phase and in clinical and endoscopic remission of UC. The identified structural changes in the gut microbiota allow for implementation as a non-invasive method for monitoring the course of progression the disease.

Introduction

Children with inflammatory bowel diseases (IBD) are particularly vulnerable to Clostridioides difficile infections (CDI) and experience significantly worse health outcomes including increased hospitalisation and shorter time to their first bowel resection. Previous studies have correlated these outcomes to specific gastrointestinal microbiome features. However, data regarding features that may predispose PIBD patients to CDI is lacking. Furthermore, it is not known whether CDI has long-term impacts on the microbiome of PIBD patients in remission.

Aims & Methods

Our primary aims were to investigate whether PIBD-CDI patients had a distinct microbiota at baseline compared to PIBD controls, to examine whether PIBD-CDI patients had a distinct microbiome during remission than those without CDI, and whether there were changes in their stool short chain fatty acids (SCFA) composition. Finally, we aimed to assess whether PIBD patients with CDI had worse outcome.
Patients aged 2-17 years and newly diagnosed with IBD between November 2014 and March 2019 were invited to participate. Each patient was requested to submit 4 stool samples in an interval of six months between samples and follow up continued until June 2021. Patients were divided into those that developed CDI (IBD-CDI) and those who did not (IBD controls). DNA was extracted from stool samples and the microbiota of each was profiled using 16S rRNA amplicon sequencing on an Illumina MiSeq. SCFA were quantified from the stool samples. Data analysis was further divided into 2 cohorts: Cohort 1 represented microbiome data from samples of patients who developed CDI and were collected prior to the infection. Cohort 2 represented microbiome data from samples of patients in remission who had CDI. Microbial composition and SCFAs were compared to non-CDI controls within each cohort. Hazard ratio (HR) was calculated to progression to biologic treatment and surgeries in patients who had a CDI compares to those who did not. Random forest classification model was used to predict CDI using microbial composition.

Results

Patients who had CDI were more likely to progress to a biologic therapy (HR 1.85 P =0.06) and had significantly more hospitalisations (RR 2.20, P< 0.001) and surgery (HR 3.15, P= 0.019).
Cohort 1:
Patients who did not develop CDI had higher abundance of Bifidobacterium bifidum, Blautia hydrogenotrophica, Clostridium sensu stricto 2 cadaveris, Eubacterium, Ruminococcus torques group, and Lachnoclostridium.
The Eubacterium, Blautia hydrogenotrophica, Ruminococcus torques group and Lachnoclostridium were associated with high levels of stool SCFAs.
Random forest classification model was able to predict patient who did not develop CDI.
Cohort 2:
Patients post CDI had significantly lower beta diversity.
The Oscillospirales CAG 352, NK4A214 group and UCG 002 were all less abundant in post-CDI patients, and were associated with higher SCFA production. Specifically, propionic acid (CAG 352), isovaleric and isobutyric acid (NK4A214 and UCG 002). Additionally, alistipes was associated with higher levels of isovaleric and isobutyric acids.

Conclusion

We were able to predict paediatric IBD patients that were protected from CDI based on baseline microbial composition. Paediatric IBD patients who developed CDI during the course of the disease developed more severe outcome with more surgeries and hospitalisations. They had long term changes to their microbiome.

References

Pascarella F, Martinelli M, Miele E, Del Pezzo M, Roscetto E, Staiano A. Impact of Clostridium difficile infection on pediatric inflammatory bowel disease. J Pediatr. 2009 Jun;154(6):854–8.
Hellmann J, Andersen H, Fei L, Linn A, Bezold R, Lake K, et al. Microbial shifts and shorter time to bowel resection surgery associated with c. difficile in pediatric Crohn’s disease. Inflamm Bowel Dis. 2020;
Sokol H, Jegou S, McQuitty C, Straub M, Leducq V, Landman C, et al. Specificities of the intestinal microbiota in patients with inflammatory bowel disease and Clostridium difficile infection. Gut Microbes. 2018;

Disclosure

No conflict of interest

Introduction

Coexistence of bile acid diarrhoea (BAD) and microscopic colitis (MC) has been described (1), and treating this with sequestrants could reduce the need for budesonide. Fasting plasma 7α-hydroxy-4-cholesten-3-one (C4) values >46 ng/mL are specific for BAD but sensitivity is only 47% (37-57%) compared with tauroselcholic [75-Se]
acid (SeHCAT) retention <10% (2). C4 values between 33 and 46 are in a diagnostic grey zone. Budesonide directly affects the bile acid homeostasis making the temporal relation between treatment and testing important. Fortunately, unlike SeHCAT testing, C4 may be sampled immediately when a patient presents with a flare of MC.

Aims & Methods

To assess the prevalence of coexisting BAD in patients with active MC.
Consequtive patients with MC and a flare fulfilling the Hjortswang diarrhoea criteria (≥3 stools or ≥1 watery stool per day) and a negative test for infectious diarrhoea were included. Patients were treated with budesonide 9 mg once daily for 6 weeks. Clinical assessment, Bristol stool charts, and fasting C4 were obtained at baseline, during the last treatment week and 4 weeks after end of treatment. Remission was defined as a daily mean of <1 watery bowel movement and <3 total bowel movements [EudraCT: 2019-002762-12].

Results

49 patients were included; 26 (53%) with collagenous colitis. Mean age was 60 (range 55-70) years, 22% current smoker, none on NSAIDs. Six (12%; 95%CI: 5–25%) of 49 patients had BAD diagnosed by C4. Another 3 patients had grey zone C4 values. Mean C4 did not change during budesonide treatment, 22 and 23 ng/mL at baseline and at week 6, respectively. SeHCAT was performed outside the study in 26 patients of whom 3 had retention of <10% and 6 of <15%. The mean total number of stools per day declined from baseline 7.9 to 1.9 and watery stools from 3.7 to 0.3 during budesonide treatment. Remission was obtained in 45. Twenty-nine had relapsed four weeks after end of budesonide treatment, 5 of 6 (83%; 95%CI: 36–100%) with MC and BAD, and 24 of 43 (56%; 95%CI: 40–71%) with MC and not BAD.

Conclusion

During an inflammatory flare of MC, 12% (5–25%) had C4-defined BAD. However, C4 only identifies about half of patients with BAD compared with the diagnostic gold standard SeHCAT, meaning about 25% of patients with MC could have coexisting BAD. Screening for BAD may have merit in MC patients with relapse of diarrhoea after budesonide treatment.

References

1 Bjørnbak C, Engel PJH, Nielsen PL, Munck LK. Microscopic colitis: clinical findings, topography, and persistence of histopathological subgroups. Aliment Pharmacol Ther. 2011, 34: 1225-34.
2. Borup C, Vinter-Jensen L, Jørgensen SPG, Wildt S, Graff J, Gregersen T, Zaremba A, Andersen TB, Nøjgaard C, Timm HB, Lamazière A, Rainteau D, Hansen SH, Rumessen JJ, Munck LK. Prospective comparison of diagnostic tests for bile acid diarrhoea. Aliment Pharmacol Ther. 2024; 59: 39-50.

Disclosure

Budesonide was donated for study use by Tillotts Pharma as an unrestricted scientific grant. Tillotts Pharma had no influence on the planning, conduct, or reporting of the study.

Introduction

Bowel urgency is one of the most bothersome symptoms of ulcerative colitis (UC) and have a great impact on quality of life and psychosocial function. The Urgency Numerical Rating Scale (U-NRS) is desinged to ojectify this subjective symptom. It measures the degree of faecal urgency experienced over the past 24 hours by patients with UC on a scale of 0-10. We assumed that the scale has a large variance, as patients evaluate their own symptoms differently, the urge may be different quality for two patients with the same score.

Aims & Methods

The objective of our study was to investigate the possible components of urgency and refine the assessment of urgency. Patients with UC older than 18 years treated at our departments for at least 3 months and having complaints were enrolled in our study. The patients filled out our non-validated questionnaire with questions about fecal urgency (U-NRS), pain level, speed of urge, incontinence and lifestyle characteristics associated with incontinence. Statistical analysis was performed using STATA 18 program.

Results

263 patients were enrolled, median age was 39.5 years, the mean of disease duration was 11.7±9.2 years. The mean U-NRS score was 3.2±3.2, 32% (84/263) had 0 score and 3% (9/263) has 10 score. 12% of the patients (n=31) felt most difficult to hold mucus, 37% (n=97) stool and 29% (n=76) other secretions. 8% (n=22) had urge with every stool in the last 24 hours, 36% (n=95) had less than half of defecations. The speed of urge defined more the level of urgency than the pain level (R2=0.798 p<0.001, β=0.894 vs R²=0.805, p<0.001, β=0.117). The duration of faeces holding in the last 7 days had significant connection to U-NRS (R²=0.211, p<0.001, β=-0.46). Patients using public restroom at least once during the last week had significantly higher score on U-NRS than other patients (4.6±3.1 vs 2.4±3 p<0.001). 12% (31/263) used diaper. The usage of diapers was also significantly more often in patients with higher U-RNS score (5.2±3.2 vs 2.9±3.1,p<0.0001). Faecal leakage both during the day and night was significiantly higher with higher U-NRS score (p<0.001 for both). Patient reported outcome (PRO2) had a weak correlation with U-NRS (R²=0.14). However if we examine bloody stool, the stool number and difficulty of faecas holding seperately we got a significant and better correlation with urgency (R²=0.43). Patients with moderately severe-severe disease (PRO2 > 5) had a higher chance to have urgency (AUC=0.9). The possibility was higher to have urgency among patients with painful defecation (R²=0.41). The level of urgency, duration of faeces holding and the frequency of faeces holding problems had the strongest correlation with U-NRS (R²=0.83). Patients defined urgency with these parameters according to this result. We created score with them, level of urgency points + half of duration points + frequency, which showed a good model to refine the definition of urgency (R²=0.83).

Conclusion

Bowel urgency affects UC patients' lives and is associated with more severe disability. Bowel urgency is common in UC affecting about two-third of patients and associates with more severe activity of UC. Fecal urgency can characterize with the level of urgency, duration of faeces holding and the frequence of stool more accurately than a single, subjective numeric scale. A new scoring system is needed to more accurately assess patients' complaints in this area and to objectify severity.

Disclosure

The authors have no potential conflict of interest to disclose

Introduction

Inflammatory bowel disease is an idiopathic disorder characterized by repeated relapses and remissions. Endoscopy, the gold standard for diagnosis and monitoring, is time consuming, expensive and invasive. Fecal calprotectin has been suggested as a surrogate marker to assess intestinal inflammation and predict disease relapse. Published evidence regarding the utility of fecal calprotectin as an indicator of clinical activity and endoscopic severity in patients with inflammatory bowel disease is not unambiguous.

Aims & Methods

The aim of this study was to determine the use of fecal calprotectin as a predictor of clinical activity and endoscopic severity in patients with inflammatory bowel disease.
Methods: Patients fulfilling selection criteria were recruited for the study. Clinical examination findings, laboratory investigations and fecal calprotectin levels were documented as per proforma. Endoscopic and clinical severity were measured by Simple Endoscopic Score and Crohn’s Disease Activity Index for Crohn’s disease and by Ulcerative Colitis Endoscopic Index of Severity score and Simple Clinical Colitis Activity Index for Ulcerative Colitis .Lab investigations and calprotectin were repeated after six months and compared with endoscopic and clinical activity indices.

Results

Fecal calprotectin correlated with Crohn’s Disease Activity Index both at first visit and follow up (r-0.689, p-0.016)(r-0.425, p-value <0.05). The sensitivity and specificity of calprotectin in detecting active disease and remission at first visit were 93.8% and 80% respectively (AUC-0.869). At follow up, the sensitivity and specificity were 80% and 93.3% respectively (AUC-0.867). In Ulcerative colitis , Simple Clinical Colitis Activity Index score correlated with fecal calprotectin levels at first visit and follow up. (r-0.231 / 0.387, p-value 0.001/0.001). The sensitivity of calprotectin in detecting active disease and remission in ulcerative colitis was 92.6% and specificity was 83.3%. AUC was 0.88. At follow up, the sensitivity of fecal calprotectin in detecting active disease and remission was 89.9% and the specificity was 87.0% ; AUC was 0.879.

Conclusion

Fecal calprotectin levels show significant correlation with clinical and endoscopic activity indices in patients of inflammatory bowel disease. Therefore, calprotectin levels in stool could be used as a surrogate marker for monitoring mucosal status as well as predicting endoscopic remission in these patients.

References

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