The children with ASD were recruited as we described in our previous study73. Briefly, ASD children were diagnosed according to the Diagnostic and Statistical Manual of Mental Disorders, 5th Edition. Each participant underwent physical, neurological, and behavioral examinations. Children were excluded from the study if they were previously diagnosed with a genetic condition such as tuberous sclerosis, significant sensory impairment, clinically evident inflammatory conditions, celiac disease, and other physiological conditions (i.e. depressive disorder, schizophrenia, and bipolar disorder). Participants that are on anti-inflammatory or anti-oxidant drugs, or take antibiotics, probiotics, prebiotics, and antifungal medications in 3 months before the feces collection were excluded. The Rome IV criteria for functional constipation were used for evaluating GI problems. The typically developed children (TD) for the fecal metabolism and cytokines measure were recruited from kindergartens through pediatricians and under physical, neurological, and behavioral examinations as the ASD group, subjects with any pathological conditions, including GI problems were excluded (Table S10).

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