Abstract
Attention-deficit/hyperactivity disorder (ADHD) is a common neurodevelopmental disorder, but the underlying pathophysiological mechanisms of ADHD remain unclear. Gut microbiota has been recognized to influence brain function and behaviors. Therefore, this study aimed to determine whether imbalanced gut microbiomes identified by a 16S rRNA sequencing approach are involved in the pathophysiology of ADHD. We recruited a total of 30 children with ADHD (mean age: 8.4 years) and a total of 30 healthy controls (mean age: 9.3 years) for this study. The dietary patterns of all participants were assessed with the food frequency questionnaire. The microbiota of fecal samples were investigated using 16S rRNA V3V4 amplicon sequencing, followed by bioinformatics and statistical analyses. We found that the gut microbiota communities in ADHD patients showed a significantly higher Shannon index and Chao index than the control subjects. Furthermore, the linear discriminant analysis effect size (LEfSe) analysis was used to identify differentially enriched bacteria between ADHD patients and healthy controls. The relative abundance of Bacteroides coprocola (B. coprocola) was decreased, while the relative abundance of Bacteroides uniformis (B. uniformis), Bacteroides ovatus (B. ovatus), and Sutterella stercoricanis (S. stercoricanis) were increased in the ADHD group. Of all participants, S. stercoricanis demonstrated a significant association with the intake of dairy, nuts/seeds/legumes, ferritin and magnesium. B. ovatus and S. stercoricanis were positively correlated to ADHD symptoms. In conclusion, we suggest that the gut microbiome community is associated with dietary patterns, and linked to the susceptibility to ADHD.
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References
Polanczyk GV, Willcutt EG, Salum GA, Kieling C, Rohde LA (2014) ADHD prevalence estimates across three decades: an updated systematic review and meta-regression analysis. Int J Epidemiol 43(2):434–442
American Psychiatric Association (2000) Diagnostic and statistical manual of mental disorders (DSM-IV-TR). American Psychiatric Association, Washington
Spencer TJ, Biederman J, Mick E (2007) Attention-deficit/hyperactivity disorder: diagnosis, lifespan, comorbidities, and neurobiology. J Pediatr Psychol 32(6):631–642
Stilling RM, Dinan TG, Cryan JF (2014) Microbial genes, brain and behaviour—epigenetic regulation of the gut–brain axis. Genes Brain Behav 13(1):69–86
Montiel-Castro AJ, Gonzalez-Cervantes RM, Bravo-Ruiseco G, Pacheco-Lopez G (2013) The microbiota-gut–brain axis: neurobehavioral correlates, health and sociality. Front Integr Neurosci 7:70
Lima-Ojeda JM, Rupprecht R, Baghai TC (2017) “I Am I and My Bacterial Circumstances”: linking gut microbiome, neurodevelopment, and depression. Front Psychiatry 8:153
Sandgren AM, Brummer RJM (2018) ADHD-originating in the gut? The emergence of a new explanatory model. Med Hypotheses 120:135–145
Yadav M, Verma MK, Chauhan NS (2018) A review of metabolic potential of human gut microbiome in human nutrition. Arch Microbiol 200(2):203–217
Borre YE, O’Keeffe GW, Clarke G, Stanton C, Dinan TG, Cryan JF (2014) Microbiota and neurodevelopmental windows: implications for brain disorders. Trends Mol Med 20(9):509–518
Cenit MC, Nuevo IC, Codoner-Franch P, Dinan TG, Sanz Y (2017) Gut microbiota and attention deficit hyperactivity disorder: new perspectives for a challenging condition. Eur Child Adolesc Psychiatry 26(9):1081–1092
Dore J, Blottiere H (2015) The influence of diet on the gut microbiota and its consequences for health. Curr Opin Biotechnol 32:195–199
Principi N, Esposito S (2016) Gut microbiota and central nervous system development. J Infect 73(6):536–546
Felice VD, O’Mahony SM (2017) The microbiome and disorders of the central nervous system. Pharmacol Biochem Behav 160:1–13
Slykerman RF, Thompson J, Waldie KE, Murphy R, Wall C, Mitchell EA (2017) Antibiotics in the first year of life and subsequent neurocognitive outcomes. Acta Paediatr 106(1):87–94
Partty A, Kalliomaki M, Wacklin P, Salminen S, Isolauri E (2015) A possible link between early probiotic intervention and the risk of neuropsychiatric disorders later in childhood: a randomized trial. Pediatr Res 77(6):823–828
Park S, Cho SC, Hong YC, Oh SY, Kim JW, Shin MS, Kim BN, Yoo HJ, Cho IH, Bhang SY (2012) Association between dietary behaviors and attention-deficit/hyperactivity disorder and learning disabilities in school-aged children. Psychiatry Res 198(3):468–476
Stevenson J (2006) Dietary influences on cognitive development and behaviour in children. Proc Nutr Soc 65(4):361–365
Sinn N (2008) Nutritional and dietary influences on attention deficit hyperactivity disorder. Nutr Rev 66(10):558–568
Millichap JG, Yee MM (2012) The diet factor in attention-deficit/hyperactivity disorder. Pediatrics 129(2):330–337
Bibbo S, Ianiro G, Giorgio V, Scaldaferri F, Masucci L, Gasbarrini A, Cammarota G (2016) The role of diet on gut microbiota composition. Eur Rev Med Pharmacol Sci 20(22):4742–4749
Wang WL, Xu SY, Ren ZG, Tao L, Jiang JW, Zheng SS (2015) Application of metagenomics in the human gut microbiome. World J Gastroenterol 21(3):803–814
Bragg L, Tyson GW (2014) Metagenomics using next-generation sequencing. Methods Mol Biol 1096:183–201
Janda JM, Abbott SL (2007) 16S rRNA gene sequencing for bacterial identification in the diagnostic laboratory: pluses, perils, and pitfalls. J Clin Microbiol 45(9):2761–2764
Caporaso JG, Lauber CL, Walters WA, Berg-Lyons D, Lozupone CA, Turnbaugh PJ, Fierer N, Knight R (2011) Global patterns of 16S rRNA diversity at a depth of millions of sequences per sample. Proc Natl Acad Sci USA 108(Suppl 1):4516–4522
Culligan EP, Marchesi JR, Hill C, Sleator RD (2014) Combined metagenomic and phenomic approaches identify a novel salt tolerance gene from the human gut microbiome. Front Microbiol 5:189
Gilbert JA, Dupont CL (2011) Microbial metagenomics: beyond the genome. Ann Rev Mar Sci 3:347–371
Oulas A, Pavloudi C, Polymenakou P, Pavlopoulos GA, Papanikolaou N, Kotoulas G, Arvanitidis C, Iliopoulos I (2015) Metagenomics: tools and insights for analyzing next-generation sequencing data derived from biodiversity studies. Bioinform Biol Insights 9:75–88
Aarts E, Ederveen THA, Naaijen J, Zwiers MP, Boekhorst J, Timmerman HM, Smeekens SP, Netea MG, Buitelaar JK, Franke B, van Hijum S, Arias Vasquez A (2017) Gut microbiome in ADHD and its relation to neural reward anticipation. PLoS One 12(9):e0183509
Jiang HY, Zhou YY, Zhou GL, Li YC, Yuan J, Li XH, Ruan B (2018) Gut microbiota profiles in treatment-naive children with attention deficit hyperactivity disorder. Behav Brain Res 347:408–413
Prehn-Kristensen A, Zimmermann A, Tittmann L, Lieb W, Schreiber S, Baving L, Fischer A (2018) Reduced microbiome alpha diversity in young patients with ADHD. PLoS One 13(7):e0200728
Kaufman J, Birmaher B, Brent D, Rao U, Flynn C, Moreci P, Williamson D, Ryan N (1997) Schedule for affective disorders and schizophrenia for school-age children-present and lifetime version (K-SADS-PL): initial reliability and validity data. J Am Acad Child Adolesc Psychiatry 36(7):980–988
Gau SF, Soong WT (1999) Psychiatric comorbidity of adolescents with sleep terrors or sleepwalking: a case-control study. Aust N Z J Psychiatry 33(5):734–739
Baron IS (2005) Test review: wechsler Intelligence Scale for Children-Fourth Edition (WISC-IV). Child Neuropsychol 11(5):471–475
Bussing R, Fernandez M, Harwood M, Wei H, Garvan CW, Eyberg SM, Swanson JM (2008) Parent and teacher SNAP-IV ratings of attention deficit hyperactivity disorder symptoms: psychometric properties and normative ratings from a school district sample. Assessment 15(3):317–328
Gau SS, Lin CH, Hu FC, Shang CY, Swanson JM, Liu YC, Liu SK (2009) Psychometric properties of the Chinese version of the Swanson, Nolan, and Pelham, Version IV Scale-Teacher Form. J Pediatr Psychol 34(8):850–861
Gau SS, Shang CY, Liu SK, Lin CH, Swanson JM, Liu YC, Tu CL (2008) Psychometric properties of the Chinese version of the Swanson, Nolan, and Pelham, version IV scale—parent form. Int J Methods Psychiatr Res 17(1):35–44
Zhang S, Faries DE, Vowles M, Michelson D (2005) ADHD Rating scale IV: psychometric properties from a multinational study as a clinician-administered instrument. Int J Methods Psychiatr Res 14(4):186–201
Lee MS, Pan WH, Liu KL, Yu MS (2006) Reproducibility and validity of a Chinese food frequency questionnaire used in Taiwan. Asia Pac J Clin Nutr 15(2):161–169
Chou WJ, Lee MF, Hou ML, Hsiao LS, Lee MJ, Chou MC, Wang LJ (2018) Dietary and nutrient status of children with attention-deficit/hyperactivity disorder: a case-control study. Asia Pac J Clin Nutr 27(6):1325–1331
Schloss PD, Westcott SL, Ryabin T, Hall JR, Hartmann M, Hollister EB, Lesniewski RA, Oakley BB, Parks DH, Robinson CJ, Sahl JW, Stres B, Thallinger GG, Van Horn DJ, Weber CF (2009) Introducing mothur: open-source, platform-independent, community-supported software for describing and comparing microbial communities. Appl Environ Microbiol 75(23):7537–7541
Kozich JJ, Westcott SL, Baxter NT, Highlander SK, Schloss PD (2013) Development of a dual-index sequencing strategy and curation pipeline for analyzing amplicon sequence data on the MiSeq Illumina sequencing platform. Appl Environ Microbiol 79(17):5112–5120
Edgar RC, Haas BJ, Clemente JC, Quince C, Knight R (2011) UCHIME improves sensitivity and speed of chimera detection. Bioinformatics 27(16):2194–2200
Wang Q, Garrity GM, Tiedje JM, Cole JR (2007) Naive Bayesian classifier for rapid assignment of rRNA sequences into the new bacterial taxonomy. Appl Environ Microbiol 73(16):5261–5267
Caporaso JG, Kuczynski J, Stombaugh J, Bittinger K, Bushman FD, Costello EK, Fierer N, Pena AG, Goodrich JK, Gordon JI, Huttley GA, Kelley ST, Knights D, Koenig JE, Ley RE, Lozupone CA, McDonald D, Muegge BD, Pirrung M, Reeder J, Sevinsky JR, Turnbaugh PJ, Walters WA, Widmann J, Yatsunenko T, Zaneveld J, Knight R (2010) QIIME allows analysis of high-throughput community sequencing data. Nat Methods 7(5):335–336
Segata N, Izard J, Waldron L, Gevers D, Miropolsky L, Garrett WS, Huttenhower C (2011) Metagenomic biomarker discovery and explanation. Genome Biol 12(6):R60
Morgan XC, Huttenhower C (2012) Chapter 12: human microbiome analysis. PLoS Comput Biol 8(12):e1002808
Bercik P, Denou E, Collins J, Jackson W, Lu J, Jury J, Deng Y, Blennerhassett P, Macri J, McCoy KD, Verdu EF, Collins SM (2011) The intestinal microbiota affect central levels of brain-derived neurotropic factor and behavior in mice. Gastroenterology 141(2):599–609 (609, e591–593)
Clarke G, Grenham S, Scully P, Fitzgerald P, Moloney RD, Shanahan F, Dinan TG, Cryan JF (2013) The microbiome-gut–brain axis during early life regulates the hippocampal serotonergic system in a sex-dependent manner. Mol Psychiatry 18(6):666–673
Sherman MP, Zaghouani H, Niklas V (2015) Gut microbiota, the immune system, and diet influence the neonatal gut–brain axis. Pediatr Res 77(1–2):127–135
Wang Y, Kasper LH (2014) The role of microbiome in central nervous system disorders. Brain Behav Immun 38:1–12
Mayer EA, Tillisch K, Gupta A (2015) Gut–brain axis and the microbiota. J Clin Invest 125(3):926–938
Wexler HM (2007) Bacteroides: the good, the bad, and the nitty-gritty. Clin Microbiol Rev 20(4):593–621
Renouf M, Hendrich S (2011) Bacteroides uniformis is a putative bacterial species associated with the degradation of the isoflavone genistein in human feces. J Nutr 141(6):1120–1126
NCBI: Bacteroides ovatus. In: Normal gut bacterium. 2019. https://www.ncbi.nlm.nih.gov/genome/?term=Bacteroides%20ovatus
Wu M, McNulty NP, Rodionov DA, Khoroshkin MS, Griffin NW, Cheng J, Latreille P, Kerstetter RA, Terrapon N, Henrissat B, Osterman AL, Gordon JI (2015) Genetic determinants of in vivo fitness and diet responsiveness in multiple human gut Bacteroides. Science 350(6256):aac5992
Coyne MJ, Roelofs KG, Comstock LE (2016) Type VI secretion systems of human gut Bacteroidales segregate into three genetic architectures, two of which are contained on mobile genetic elements. BMC Genomics 17:58
Kitahara M, Sakamoto M, Ike M, Sakata S, Benno Y (2005) Bacteroides plebeius sp. nov. and Bacteroides coprocola sp. nov., isolated from human faeces. Int J Syst Evol Microbiol 55(Pt 5):2143–2147
Tillisch K, Mayer EA, Gupta A, Gill Z, Brazeilles R, Le Neve B, van Hylckama Vlieg JET, Guyonnet D, Derrien M, Labus JS (2017) Brain structure and response to emotional stimuli as related to gut microbial profiles in healthy women. Psychosom Med 79(8):905–913
Greetham HL, Collins MD, Gibson GR, Giffard C, Falsen E, Lawson PA (2004) Sutterella stercoricanis sp. nov., isolated from canine faeces. Int J Syst Evol Microbiol 54(Pt 5):1581–1584
Williams BL, Hornig M, Parekh T, Lipkin WI (2012) 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. MBio 3(1):e00261
Wang LJ, Yu YH, Fu ML, Yeh WT, Hsu JL, Yang YH, Chen WJ, Chiang BL, Pan WH (2018) Attention deficit-hyperactivity disorder is associated with allergic symptoms and low levels of hemoglobin and serotonin. Sci Rep 8(1):10229
Chou WJ, Lee MF, Hou ML, Hsiao LS, Lee MJ, Chou MC, Wang LJ (2018) Dietary and nutrient status of children with attention-deficit/hyperactivity disorder: a case-control study. Asia Pac J Clin Nutr 27(6):1325–1331
Proctor C, Thiennimitr P, Chattipakorn N, Chattipakorn SC (2017) Diet, gut microbiota and cognition. Metab Brain Dis 32(1):1–17
Diaz Heijtz R, Wang S, Anuar F, Qian Y, Bjorkholm B, Samuelsson A, Hibberd ML, Forssberg H, Pettersson S (2011) Normal gut microbiota modulates brain development and behavior. Proc Natl Acad Sci USA 108(7):3047–3052
Hopkins MJ, Sharp R, Macfarlane GT (2002) Variation in human intestinal microbiota with age. Dig Liver Dis 34(Suppl 2):S12–S18
Axelsson PB, Clausen TD, Petersen AH, Hageman I, Pinborg A, Kessing LV, Bergholt T, Rasmussen SC, Keiding N, Lokkegaard ECL (2019) Investigating the effects of cesarean delivery and antibiotic use in early childhood on risk of later attention deficit hyperactivity disorder. J Child Psychol Psychiatry 60(2):151–159
Liu YCLS, Shang CY, Lin CH, Tu CL, Gau SF (2006) Norm of the Chinese version of the Swanson, Nolan and Pelham, version IV scale for ADHD. Taiwan J Psychiatry 20:290–304
Jain A, Li XH, Chen WN (2018) Similarities and differences in gut microbiome composition correlate with dietary patterns of Indian and Chinese adults. AMB Express 8(1):104
Acknowledgements
The authors would like to thank Professor Wei-Tsun Soong for granting us the use of the Chinese version of the K-SADS, and Professor Shur-Fen Gau for granting our use of the Chinese version of the SNAP-IV.
Funding
This work was supported by grant from the Chang Gung Memorial Hospital Research Grant (CMRPG8E1441) and the Taiwan Ministry of Science and Technology (MOST 107-2628-B-182-001).
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Supplementary material 1 The distribution of enriched bacteria identified at genus level in the ADHD patients and healthy controls using LEfSe analysis. Linear discriminant analysis (LDA) plots at genus levels showed the enriched bacteria in ADHD patients and healthy controls (JPEG 225 kb)
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Wang, LJ., Yang, CY., Chou, WJ. et al. Gut microbiota and dietary patterns in children with attention-deficit/hyperactivity disorder. Eur Child Adolesc Psychiatry 29, 287–297 (2020). https://doi.org/10.1007/s00787-019-01352-2
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DOI: https://doi.org/10.1007/s00787-019-01352-2
Keywords
- ADHD
- Gut–brain axis
- 16S rRNA sequencing
- Microbiome
- Biomarker