Differences in fecal microbial metabolites and microbiota of children with autism spectrum disorders

Kang, Dae-Wook; Ilhan, Zehra Esra; Isern, Nancy G.; Hoyt, David W.; Howsmon, Daniel P.; Shaffer, Michael; Lozupone, Catherine A.; Hahn, Juergen; Adams, James B.; Krajmalnik-Brown, Rosa

doi:10.1016/j.anaerobe.2017.12.007

Differences in fecal microbial metabolites and microbiota of children with autism spectrum disorders

Journal Article · 01 February 2018 · Anaerobe

DOI:https://doi.org/10.1016/j.anaerobe.2017.12.007· OSTI ID:1422278

Kang, Dae-Wook; Ilhan, Zehra Esra; Isern, Nancy G.; ; ; Shaffer, Michael; Lozupone, Catherine A.; Hahn, Juergen; ;

Evidence supporting that gut problems are linked to ASD symptoms has been accumulating both in humans and animal models of ASD. Gut microbes and their metabolites may be linked not only to GI problems but also to ASD behavior symptoms. Despite this high interest, most previous studies have looked mainly at microbial structure, and studies on fecal metabolites are rare in the context of ASD. Thus, we aimed to detect fecal metabolites that may be present at significantly different concentrations between 21 children with ASD and 23 neurotypical children and to investigate its possible link to human gut microbiome. Using NMR spectroscopy and 16S rRNA gene amplicon sequencing, we examined metabolite profiles and microbial compositions in fecal samples, respectively. Of the 59 metabolites detected, isopropanol concentrations were significantly higher in feces of children with ASD after multiple testing corrections. We also observed similar trends of fecal metabolites to previous studies; children with ASD have higher fecal p-cresol and possibly lower GABA concentrations. In addition, Fisher Discriminant Analysis (FDA) with leave-out-validation suggested that a group of metabolites- caprate, nicotinate, glutamine, thymine, and aspartate- may potentially function as a biomarker to separate ASD participants from the neurotypical group (78% sensitivity and 81% specificity). Consistent with our previous Arizona cohort study, we also confirmed lower gut microbial diversity and reduced relative abundances of Prevotella copri in children with ASD. After multiple testing corrections, we also learned that relative abundances of Feacalibacterium prausnitzii and Haemophilus parainfluenzae were lower in feces of children with ASD. Despite a relatively short list of fecal metabolites, the data in this study support that children with ASD have altered metabolite profiles in feces when compared with neurotypical children and warrant further investigation of metabolites in larger cohorts.

Research Organization:: Pacific Northwest National Laboratory (PNNL), Richland, WA (United States). Environmental Molecular Sciences Laboratory (EMSL)

Sponsoring Organization:: USDOE

DOE Contract Number:: AC05-76RL01830

OSTI ID:: 1422278

Report Number(s):: PNNL-SA-127135; 49112; 48610; WN0219080

Journal Information:: Anaerobe, Vol. 49, Issue C; ISSN 1075-9964

Publisher:: Elsevier

Country of Publication:: United States

Language:: English

References (77)

Autism No authors listed The Lancet https://doi.org/10.1016/S0140-6736(13)61539-1		March 2014
The Central Nervous System and the Gut Microbiome No authors listed Cell https://doi.org/10.1016/j.cell.2016.10.027		November 2016
Maternal gut bacteria promote neurodevelopmental abnormalities in mouse offspring No authors listed Nature https://doi.org/10.1038/nature23910		September 2017
Evaluation, Diagnosis, and Treatment of Gastrointestinal Disorders in Individuals With ASDs: A Consensus Report No authors listed Pediatrics https://doi.org/10.1542/peds.2009-1878C		January 2010
Gastrointestinal Symptoms in Autism Spectrum Disorder: A Meta-analysis No authors listed PEDIATRICS https://doi.org/10.1542/peds.2013-3995		April 2014
Impaired Carbohydrate Digestion and Transport and Mucosal Dysbiosis in the Intestines of Children with Autism and Gastrointestinal Disturbances No authors listed PLoS ONE https://doi.org/10.1371/journal.pone.0024585		September 2011
Gastrointestinal microbiota in children with autism in Slovakia No authors listed Physiology & Behavior https://doi.org/10.1016/j.physbeh.2014.10.033		January 2015
Pyrosequencing study of fecal microflora of autistic and control children No authors listed Anaerobe https://doi.org/10.1016/j.anaerobe.2010.06.008		August 2010
Real-Time PCR Quantitation of Clostridia in Feces of Autistic Children No authors listed Applied and Environmental Microbiology https://doi.org/10.1128/AEM.70.11.6459-6465.2004		November 2004
Differences between the gut microflora of children with autistic spectrum disorders and that of healthy children No authors listed Journal of Medical Microbiology https://doi.org/10.1099/jmm.0.46101-0		October 2005
Gastrointestinal Microflora Studies in Late‐Onset Autism No authors listed Clinical Infectious Diseases https://doi.org/10.1086/341914		September 2002
Fecal Microbiota and Metabolome of Children with Autism and Pervasive Developmental Disorder Not Otherwise Specified No authors listed PLoS ONE https://doi.org/10.1371/journal.pone.0076993		October 2013
Low Relative Abundances of the Mucolytic Bacterium Akkermansia muciniphila and Bifidobacterium spp. in Feces of Children with Autism No authors listed Applied and Environmental Microbiology https://doi.org/10.1128/AEM.05212-11		July 2011
Application of Novel PCR-Based Methods for Detection, Quantitation, and Phylogenetic Characterization of Sutterella Species in Intestinal Biopsy Samples from Children with Autism and Gastrointestinal Disturbances No authors listed mBio https://doi.org/10.1128/mBio.00261-11		January 2012
Comparison of Fecal Microbiota in Children with Autism Spectrum Disorders and Neurotypical Siblings in the Simons Simplex Collection No authors listed PLOS ONE https://doi.org/10.1371/journal.pone.0137725		October 2015
Molecular Characterisation of Gastrointestinal Microbiota of Children With Autism (With and Without Gastrointestinal Dysfunction) and Their Neurotypical Siblings: GI microbiota of children with autism No authors listed Autism Research https://doi.org/10.1002/aur.1253		September 2012
Microbiota Modulate Behavioral and Physiological Abnormalities Associated with Neurodevelopmental Disorders No authors listed Cell https://doi.org/10.1016/j.cell.2013.11.024		December 2013
Effects of the enteric bacterial metabolic product propionic acid on object-directed behavior, social behavior, cognition, and neuroinflammation in adolescent rats: Relevance to autism spectrum disorder No authors listed Behavioural Brain Research https://doi.org/10.1016/j.bbr.2010.10.005		February 2011
Microbial Reconstitution Reverses Maternal Diet-Induced Social and Synaptic Deficits in Offspring No authors listed Cell https://doi.org/10.1016/j.cell.2016.06.001		June 2016
Ingestion of Lactobacillus strain regulates emotional behavior and central GABA receptor expression in a mouse via the vagus nerve No authors listed Proceedings of the National Academy of Sciences https://doi.org/10.1073/pnas.1102999108		August 2011
Pre- and Neonatal Exposure to Lipopolysaccharide or the Enteric Metabolite, Propionic Acid, Alters Development and Behavior in Adolescent Rats in a Sexually Dimorphic Manner No authors listed PLoS ONE https://doi.org/10.1371/journal.pone.0087072		January 2014
Oxytocin, Vasopressin, and the Neurogenetics of Sociality No authors listed Science https://doi.org/10.1126/science.1158668		November 2008
Can oxytocin treat autism? No authors listed Science https://doi.org/10.1126/science.aaa8120		February 2015
Effect of a vitamin/mineral supplement on children and adults with autism No authors listed BMC Pediatrics https://doi.org/10.1186/1471-2431-11-111		December 2011
Increased serum levels of glutamate in adult patients with autism No authors listed Progress in Neuro-Psychopharmacology and Biological Psychiatry https://doi.org/10.1016/j.pnpbp.2006.06.013		December 2006
Metabolic Perturbance in Autism Spectrum Disorders: A Metabolomics Study No authors listed Journal of Proteome Research https://doi.org/10.1021/pr300910n		October 2012
Urinary Metabolic Phenotyping Differentiates Children with Autism from Their Unaffected Siblings and Age-Matched Controls No authors listed Journal of Proteome Research https://doi.org/10.1021/pr901188e		June 2010
Urinary p -cresol is elevated in young French children with autism spectrum disorder: a replication study No authors listed Biomarkers https://doi.org/10.3109/1354750X.2014.936911		May 2014
Urinary metabolomics of young Italian autistic children supports abnormal tryptophan and purine metabolism No authors listed Molecular Autism https://doi.org/10.1186/s13229-016-0109-5		November 2016
Elevated Fecal Short Chain Fatty Acid and Ammonia Concentrations in Children with Autism Spectrum Disorder No authors listed Digestive Diseases and Sciences https://doi.org/10.1007/s10620-012-2167-7		April 2012
Evaluation of the bacterial diversity among and within individual venous leg ulcers using bacterial tag-encoded FLX and Titanium amplicon pyrosequencing and metagenomic approaches No authors listed BMC Microbiology https://doi.org/10.1186/1471-2180-9-226		January 2009
QIIME allows analysis of high-throughput community sequencing data No authors listed Nature Methods https://doi.org/10.1038/nmeth.f.303		April 2010
Search and clustering orders of magnitude faster than BLAST No authors listed Bioinformatics https://doi.org/10.1093/bioinformatics/btq461		August 2010
Chimeric 16S rRNA sequence formation and detection in Sanger and 454-pyrosequenced PCR amplicons No authors listed Genome Research https://doi.org/10.1101/gr.112730.110		January 2011
The Ribosomal Database Project: improved alignments and new tools for rRNA analysis No authors listed Nucleic Acids Research https://doi.org/10.1093/nar/gkn879		January 2009
Conservation evaluation and phylogenetic diversity No authors listed Biological Conservation https://doi.org/10.1016/0006-3207(92)91201-3		January 1992
UniFrac: a New Phylogenetic Method for Comparing Microbial Communities No authors listed Applied and Environmental Microbiology https://doi.org/10.1128/AEM.71.12.8228-8235.2005		December 2005
Predictive functional profiling of microbial communities using 16S rRNA marker gene sequences No authors listed Nature Biotechnology https://doi.org/10.1038/nbt.2676		August 2013
KEGG: new perspectives on genomes, pathways, diseases and drugs No authors listed Nucleic Acids Research https://doi.org/10.1093/nar/gkw1092		November 2016
Cytoscape: A Software Environment for Integrated Models of Biomolecular Interaction Networks No authors listed Genome Research https://doi.org/10.1101/gr.1239303		November 2003
Isopropanol poisoning No authors listed Clinical Toxicology https://doi.org/10.3109/15563650.2014.914527		May 2014
Can Alcohol-Based Hand-Rub Solutions Cause You To Lose Your Driver's License? Comparative Cutaneous Absorption of Various Alcohols No authors listed Antimicrobial Agents and Chemotherapy https://doi.org/10.1128/AAC.01320-06		March 2007
Toxic Alcohol Ingestions: Clinical Features, Diagnosis, and Management No authors listed Clinical Journal of the American Society of Nephrology https://doi.org/10.2215/CJN.03220807		November 2007
Treatment of the alcohol intoxications: ethylene glycol, methanol and isopropanol No authors listed Current Opinion in Nephrology and Hypertension https://doi.org/10.1097/00041552-200011000-00017		January 2000
Studies on the Organism Producing Isopropanol from Acetone. Part IV [アセトンを還元してイソプロパノールを生成する細菌に就て(第6報)] No authors listed Journal of the agricultural chemical society of Japan https://doi.org/10.1271/nogeikagaku1924.34.7_616		January 1960
Alcohol dehydrogenase: multiplicity and relatedness in the solvent-producing clostridia No authors listed FEMS Microbiology Reviews https://doi.org/10.1111/j.1574-6976.1995.tb00210.x		October 1995
Acetone, Isopropanol, and Butanol Production by Clostridium beijerinckii (syn. Clostridium butylicum) and Clostridium aurantibutyricum No authors listed Applied and Environmental Microbiology https://doi.org/10.1128/AEM.45.3.1160-1163.1983		January 1983
Acetone-butanol-isopropanol production byClostridium beijerinckii (synonym,Clostridium butylicum) No authors listed Biotechnology Letters https://doi.org/10.1007/BF01040869		May 1986
Urinary p-cresol in autism spectrum disorder No authors listed Neurotoxicology and Teratology https://doi.org/10.1016/j.ntt.2012.09.002		March 2013
p -Hydroxyphenylacetate decarboxylase from Clostridium difficile : A novel glycyl radical enzyme catalysing the formation of No authors listed European Journal of Biochemistry https://doi.org/10.1046/j.1432-1327.2001.02001.x		March 2001
Resistant starch reduces colonic and urinary p -cresol in rats fed a tyrosine-supplemented diet, whereas konjac mannan does not No authors listed Bioscience, Biotechnology, and Biochemistry https://doi.org/10.1080/09168451.2016.1194183		October 2016
The uremic solutes p-cresol and indoxyl sulfate inhibit endothelial proliferation and wound repair No authors listed Kidney International https://doi.org/10.1111/j.1523-1755.2004.00399.x		February 2004
Effects of intestinal bacteria-derived p-cresyl sulfate on Th1-type immune response in vivo and in vitro No authors listed Toxicology and Applied Pharmacology https://doi.org/10.1016/j.taap.2013.10.016		January 2014
Distinct Urinary Metabolic Profile of Human Colorectal Cancer No authors listed Journal of Proteome Research https://doi.org/10.1021/pr201001a		December 2011
The deleterious metabolic and genotoxic effects of the bacterial metabolite p-cresol on colonic epithelial cells No authors listed Free Radical Biology and Medicine https://doi.org/10.1016/j.freeradbiomed.2015.04.004		August 2015
The enteric bacterial metabolite propionic acid alters brain and plasma phospholipid molecular species: further development of a rodent model of autism spectrum disorders No authors listed Journal of Neuroinflammation https://doi.org/10.1186/1742-2094-9-153		July 2012
Butyrate Enhances the Intestinal Barrier by Facilitating Tight Junction Assembly via Activation of AMP-Activated Protein Kinase in Caco-2 Cell Monolayers No authors listed The Journal of Nutrition https://doi.org/10.3945/jn.109.104638		July 2009
Potential beneficial effects of butyrate in intestinal and extraintestinal diseases No authors listed World Journal of Gastroenterology https://doi.org/10.3748/wjg.v17.i12.1519		January 2011
Epigenetic Factors and Autism Spectrum Disorders No authors listed NeuroMolecular Medicine https://doi.org/10.1007/s12017-013-8222-5		March 2013
GABAergic/glutamatergic imbalance relative to excessive neuroinflammation in autism spectrum disorders No authors listed Journal of Neuroinflammation https://doi.org/10.1186/s12974-014-0189-0		November 2014
Role of octanoic and decanoic acids in the control of seizures. No authors listed Archives of Disease in Childhood https://doi.org/10.1136/adc.61.12.1173		December 1986
Seizure control by decanoic acid through direct AMPA receptor inhibition No authors listed Brain https://doi.org/10.1093/brain/awv325		November 2015
NMDA receptors and ligands in the vertebrate CNS No authors listed Progress in Neurobiology https://doi.org/10.1016/0301-0082(88)90027-5		January 1988
NMDA receptor dysfunction in autism spectrum disorders No authors listed Current Opinion in Pharmacology https://doi.org/10.1016/j.coph.2014.10.007		February 2015
Roles of glutamine in neurotransmission No authors listed Neuron Glia Biology https://doi.org/10.1017/S1740925X11000093		November 2010
Serotonin: a mediator of the brain-gut connection No authors listed The American Journal of Gastroenterology https://doi.org/10.1111/j.1572-0241.2000.03177.x		October 2000
Microbiota Transfer Therapy alters gut ecosystem and improves gastrointestinal and autism symptoms: an open-label study No authors listed Microbiome https://doi.org/10.1186/s40168-016-0225-7		January 2017
Modeling environmental risk factors of autism in mice induces IBD-related gut microbial dysbiosis and hyperserotonemia No authors listed Molecular Brain https://doi.org/10.1186/s13041-017-0292-0		April 2017
Faecalibacterium prausnitzii is an anti-inflammatory commensal bacterium identified by gut microbiota analysis of Crohn disease patients No authors listed Proceedings of the National Academy of Sciences https://doi.org/10.1073/pnas.0804812105		October 2008
Identification of an anti-inflammatory protein from Faecalibacterium prausnitzii , a commensal bacterium deficient in Crohn’s disease No authors listed Gut https://doi.org/10.1136/gutjnl-2014-307649		June 2015
Global and Deep Molecular Analysis of Microbiota Signatures in Fecal Samples From Patients With Irritable Bowel Syndrome No authors listed Gastroenterology https://doi.org/10.1053/j.gastro.2011.07.043		November 2011
Abnormal intestinal permeability in children with autism No authors listed Acta Paediatrica https://doi.org/10.1111/j.1651-2227.1996.tb14220.x		September 1996
Alterations of the Intestinal Barrier in Patients With Autism Spectrum Disorders and in Their First-degree Relatives No authors listed Journal of Pediatric Gastroenterology & Nutrition https://doi.org/10.1097/MPG.0b013e3181dcc4a5		January 2010
Haemophilus parainfluenzae as a marker of the upper respiratory tract microbiota changes under the influence of preoperative prophylaxis with or without postoperative treatment in patients with lung cancer No authors listed BMC Microbiology https://doi.org/10.1186/s12866-016-0679-6		April 2016
Immune Response to Haemophilus parainfluenzae in Patients with Chronic Obstructive Lung Disease No authors listed Clinical Diagnostic Laboratory Immunology https://doi.org/10.1128/CDLI.7.1.25-30.2000		January 2000
Dynamics and Stabilization of the Human Gut Microbiome during the First Year of Life No authors listed Cell Host & Microbe https://doi.org/10.1016/j.chom.2015.04.004		May 2015
Gastrointestinal Microbiome Signatures of Pediatric Patients With Irritable Bowel Syndrome No authors listed Gastroenterology https://doi.org/10.1053/j.gastro.2011.06.072		November 2011

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Rett Syndrome and Other Neurodevelopmental Disorders Share Common Changes in Gut Microbial Community: A Descriptive Review No authors listed International Journal of Molecular Sciences https://doi.org/10.3390/ijms20174160		August 2019
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Tobacco use and cessation in psychiatric disorders: National Institute of Mental Health report No authors listed Nicotine & Tobacco Research https://doi.org/10.1080/14622200802443569		December 2008
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