Rotaviruses, causing acute gastroenteritis, that infect humans and animals around the world. There are many assays had been developed for the detection of rotavirus or the viral antigens. The present study was done on 79 samples of stool collected from pediatric patients with acute watery diarrhea aged from one months to 5 years admitted to Basrah Maternity and children hospital in Basrah province, during the period from October 2014 to February 2015. Ninety diarrheic fecal bovine samples were included in this study. All samples were used for the investigation and detection of rotavirus antigen by Enzyme-Linked Immunosorbent assay (ELISA).According to ELISA results, 10 out of 79(12.7%) pediatric stool samples rotavirus antigens were detected in children. Percentage (20.7%) of positive rotavirus antigen were detected in the patients at second age group (>6 months). Followed by 8% of patients at first age group (<6 months) these differences were not significant (P>0.05). The percentage of rotavirus antigen was higher in males patients (16.7%) compared to females (P>0.05) and also the differences were not significant differences (P>0.05). These results of rotavirus antigen detection in 90 diarrheic bovine fecal samples showed that this antigen was excreted by 56.7%of diarrheic calves. Additionally the higher non-significant (P>0.05) excretion percentage according to age was observed in 63.4 % of calves > 1 year old and the lower percentage(51.1%) was observed in the first age group( < 1year) calves old. The differences in sex were not significant (P>0.05) in the percentage of rotavirus antigen detection were also detected as 63.5% of male fecal samples show positive rotavirus antigen excretion whereas only 47.4% of female fecal samples were positive.