Objectives: The objectives of this study were to: Indentify nutrient intake deficiencies in children with ASD; Describe gastrointestinal symptoms in children with ASD; Characterize (via molecular techniques) the stool microflora (beneficial and pathogenic) in children with ASD; and Analyze for relationships among these three variables to explore trends for a possible clinical phenotype for further hypothesis testing.
Methods: Twenty-four children with ASD and 12 typical children were recruited for this pilot study. Nutrient Intake, Gastrointestinal Symptoms, and stool microflora were measured in the ASD children. Stool samples from children with ASD were age and gender-matched with samples from typical children. Three day food records were collected and analyzed using the Food Processor Nutrient Analysis Software, 2009 (ESHA Research, Salem, OR). The degree to which the children experienced the various symptoms (bloating, flatulence, abdominal pain, diarrhea, headache) was recorded by placing an “X” on a 5 cm. line numbered 0-5 on a record sheet, with 0= none and 5 = severe. Stool was collected and handled as described in a recently published OSU protocol which demonstrated improved PCR—Polymerase chain reaction - quality community DNA extraction. The DNA extraction called for the repeated bead-beating plus column (RBB+C) method. For Bifidobacteria, Lactobacilli, and Clostridia, the specific primers for each real-time PCR assay were used.
Results: A deficient nutrient intake was considered as such for <80% of the Dietary Reference Intake (DRI) of the specific vitamin or mineral. Following is the percentage of the children for who had deficient intakes for each nutrient: Vitamin A- 67%; Biotin- 63%;Calcium 37%; Choline-96%; Folic acid- 25%)Vitamin D, 54%; Vitamin E- 67%, Vitamin K-92%; Magnesium-42%; Manganese-63%; and Potassium-71%. Forty-six percent (11/24) of the children regularly experienced at least one gastrointestinal symptom, with Bloating, Flatulence, and Constipation being reported more frequently as 2.5-4.0 (moderately severe), and Abdominal Pain and Diarrhea reported as mild (1.0-2.0). Preliminary molecular characterization of stool reveals a statistically significant lower count of Lactobacilli (P= 0.0081) in the children with ASD vs. typical children, and no difference in the Bifidobacteria enumeration. Pathogenic bacteria are currently being analyzed. Relationships among the variables will be statistically tested.
Conclusions: The majority of children with ASD have compromised nutrient intakes. Half of the children regularly experience gastrointestinal symptoms, with bloating, flatulence, and constipation being the most severe. The beneficial bacteria, Lactobacilli, is significantly lower in the gastrointestinal tracts of children with autism in this study. Relationships among the variables, implications, and preliminary discussions regarding possible clinical phenotypes will be further explored.