Evelien F. de Groot

Faecal gas analysis by electronic nose as novel, non-invasive method for assessment of active and quiescent paediatric inflammatory bowel disease: Proof of principle study.

BACKGROUND AND AIMS Inflammatory bowel disease (IBD) and its two phenotypes ulcerative colitis (UC) and Crohns disease (CD) are essentially assessed by endoscopy, both in initial diagnostic work-up and during follow-up. This carries a high burden, especially on paediatric patients. Faecal volatile organic compounds (VOCs) are considered potential non-invasive biomarkers for intestinal diseases linked to gut microbiota alterations. We hypothesized that faecal VOC analysis by electronic nose allows discrimination of children with CD, UC and controls during active disease and remission. METHODS Faecal VOC patterns of children with newly diagnosed IBD and controls were studied by an electronic nose (Cyranose 320®), at baseline and upon achieving remission at 6-weeks of follow-up. Disease activity was assessed by global physicians assessment, substantiated by serum C-reactive protein and faecal calprotectin. Internally cross-validated receiver-operator-characteristic curves and corresponding sensitivity and specificity for detection of IBD were calculated RESULTS: Faecal VOC profiles of patients with UC (26) and CD (29) differed from controls (28); in active disease (AUC±95% CI, p-value, sensitivity, specificity: 1.00±0.00; p<0.001, 100%, 100%) and (0.85±0.05, p<0.001, 86%, 67%) and in clinical remission (0.94±0.06, p<0.001, 94%, 94%) and (0.94±0.06, p<0.001, 94%, 94%), respectively. Furthermore, CD-patients differed from UC-patients during active disease (0.96±0.03; p<0.001, 97%, 92%), and upon achieving clinical remission (0.81±0.08, p=0.002, 88%, 72%). CONCLUSION Faecal VOC analysis allowed discrimination of paediatric patients with IBD from controls, both during active disease and remission. It therefore has potential as non-invasive test, in both diagnostic work-up and assessment of disease activity in IBD.

Composition and stability of intestinal microbiota of healthy children within a Dutch population

Numerous diseases linked to microbial imbalance can be traced back to childhood, illustrating the impact of the juvenile microbiota development from infancy toward adulthood. However, knowledge on this subject is currently very limited. The primary aim of this study was to characterize composition and short‐ and long‐term stability of the intestinal microbiota in healthy children. Between November 2011 and June 2014, 61 children 2 to 18 yr of age from different areas in The Netherlands were included and instructed to collect fecal samples weekly, for 6 wk, and a follow‐up sample after 18 mo. The intergenic spacer profiling technique (IS‐pro) was used to analyze all available fecal samples. Microbial diversity was calculated by the Shannon diversity index and individual compositional stability by comparing all collection time points. Microbial stability varied per phylum (P < 0.0005), declined rapidly in a short time period, and subsequently stabilized on the long run with very gradual variation, leading to an overall compositional stability of 70% on average over a period of 18 mo. Higher species diversity was correlated to a higher overall compositional stability (P< 0.001). We observed an age‐independent bacterial shared core consisting of a limited number of species. In conclusion, in this study, we showed that microbial composition stability in children varied per phylum, at both short‐term and long‐term intervals. Healthy children seem to share a microbiome core consisting of a limited number of species.—De Meij, T. G. J., Budding, A. E., de Groot, E. F. J., Jansen, F. M., Kneepkens, C. M. F., Benninga, M. A., Penders, J., van Bodegraven, A. A., Savelkoul, P. H. M. Composition and stability of intestinal microbiota of healthy children within a Dutch population. FASEB J. 30, 1512–1522 (2016). www.fasebj.org

Characterization of Microbiota in Children with Chronic Functional Constipation

Objectives Disruption of the intestinal microbiota is considered an etiological factor in pediatric functional constipation. Scientifically based selection of potential beneficial probiotic strains in functional constipation therapy is not feasible due to insufficient knowledge of microbiota composition in affected subjects. The aim of this study was to describe microbial composition and diversity in children with functional constipation, compared to healthy controls. Study Design Fecal samples from 76 children diagnosed with functional constipation according to the Rome III criteria (median age 8.0 years; range 4.2–17.8) were analyzed by IS-pro, a PCR-based microbiota profiling method. Outcome was compared with intestinal microbiota profiles of 61 healthy children (median 8.6 years; range 4.1–17.9). Microbiota dissimilarity was depicted by principal coordinate analysis (PCoA), diversity was calculated by Shannon diversity index. To determine the most discriminative species, cross validated logistic ridge regression was performed. Results Applying total microbiota profiles (all phyla together) or per phylum analysis, no disease-specific separation was observed by PCoA and by calculation of diversity indices. By ridge regression, however, functional constipation and controls could be discriminated with 82% accuracy. Most discriminative species were Bacteroides fragilis, Bacteroides ovatus, Bifidobacterium longum, Parabacteroides species (increased in functional constipation) and Alistipes finegoldii (decreased in functional constipation). Conclusions None of the commonly used unsupervised statistical methods allowed for microbiota-based discrimination of children with functional constipation and controls. By ridge regression, however, both groups could be discriminated with 82% accuracy. Optimization of microbiota-based interventions in constipated children warrants further characterization of microbial signatures linked to clinical subgroups of functional constipation.

Sniffing Out Paediatric Gastrointestinal Diseases: The Potential of Volatile Organic Compounds as Biomarkers for Disease

ABSTRACT The diagnostic work-up and follow-up of paediatric functional gastrointestinal disorders and organic conditions usually includes invasive tests, carrying a high burden on patients. There is a place, therefore, for novel, noninvasive disease-specific biomarkers. Volatile organic compounds (VOCs), originating from (patho)physiological metabolic processes in the human body, are excreted as waste products through all conceivable bodily excrements. The spectrum of VOCs harbours a magnificent source of information, with the potential to serve as noninvasive diagnostic biomarkers and to monitor disease activity. VOC analysis has been studied in children and infants with a variety of gastrointestinal diseases, including inflammatory bowel disease, liver diseases, irritable bowel syndrome, necrotizing enterocolitis and infectious diarrhoea. Most of these studies, although limited in sample size, show that patients can be discriminated from controls based on their VOC profiles, underscoring the potential of VOC analysis in diagnosis and follow-up. Currently, however, the application of VOC analysis in clinical practice is limited; substantial challenges, including methodological, biological, and analytical problems, still need to be met. In this review we provide an overview of the available literature on the potential of VOCs as biomarkers for paediatric gastrointestinal diseases. We discuss the available techniques to analyse VOCs and provide topics for VOC-related research, which need to be addressed before VOC diagnostics can be implemented in daily clinical practice.

Su1268 Microbiota Profiling in Children With Functional Constipation

BACKGROUND: The prevalence of functional gastrointestinal disorders (FGIDs) in infants and toddlers by Rome III criteria is unknown. The purpose of the current study was to determine the U.S. prevalence of gastrointestinal symptoms suggestive of an infant/toddler FGID as reported by mothers in a representative community sample. METHODS: A national community sample provided by the market research company Cint USA, Inc. consisted of 1,447 mothers of children age 0-18 from all 50 US states who completed an internet survey including Rome III symptom questions, demographics, health history and the pediatric quality of life questionnaire. Of these, 320 provided information about children 0-3 years old and 264 (82.5%) were judged valid (consistent survey answers and no diagnosed disease explaining the GI symptoms). Stratified sampling was used to ensure equal gender proportions, child age distribution and adequate representation of Hispanic and African American children. To avoid biasing the sample, invitations to participate described the study as a general health survey rather than a survey of gastrointestinal symptoms. RESULTS: The children in the sample were 47.7% female, 26.7% Hispanic, 56.8% Caucasian, 12% African American, and 6% Asian; 99.2% lived full time with their mothers. Children were approximately equally divided by age (32%

Flatography: Detection of gastrointestinal diseases by faecal gas analysis.

100,000 (9%). Twenty seven percent of infants/toddlers qualified for at least one FGID by Rome III criteria. All seven infant/toddler FGIDs occurred, with infant regurgitation as the most common disorder (see Table). No race and gender differences were found. Compared to those who did not meet Rome criteria, toddlers with FGID had lower quality of life (M=80.1 vs M=90.3, p<.001). There was a trend for children to qualify for a FGID if their mother also qualified for an FGID (p = .08). A child was also more likely to have hard stools if the parent reported hard stools (p= 0.02) but no such association was found for loose stools. CONCLUSION: This first national survey in the U.S. on infant and toddler FGIDS shows that these disorders are common, can be identified in the general population and reduce quality of life. No gender and race differences were found. More research is needed into these conditions as it may improve the lives of a significant number of young children [Research supported by the Rome foundation]. Percentages of infants/toddlers meeting Rome III criteria by mothers report