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Background: Some studies have shown that low vitamin D is a risk factor for infectious diarrhea, but some have rejected it. Objectives: Considering the high prevalence of infectious diarrhea among children, the high prevalence of vitamin D deficiency, and the possible mutual effect of these two, the present study aimed to measure vitamin D in children with acute diarrhea. Methods: This study enrolled 222 children aged 2 - 14 in healthy control and acute diarrhea groups. The 25-hydroxyvitamin D (25(OH)D) level was measured in all samples by the Chemiluminescent Immunoassays (CLIA) method. Results: The mean age of the participants was 5 ± 2.7 years. Patients with acute diarrhea had lower vitamin D than healthy controls (P = 0.04). The frequency of deficient and insufficient vitamin D levels was higher in the acute diarrhea group than in the healthy control group, but insignificantly (P = 0.146). Conclusions: The present study revealed an association between insufficient vitamin D and acute diarrhea. Hence, low vitamin D is a risk factor for acute diarrhea.