Enteric adenoviruses are important etiological agents associated with sporadic infections and outbreaks of acute gastroenteritis in infants and children. Fecal samples were collected from 439 hospitalized patients in the years 2005-2007 from Pune, Aurangabad, and Nagpur cities of western India to identify the most prevalent strains of enteric adenoviruses. The viruses were detected by PCR and characterized by sequencing and phylogenetic analysis. The prevalence of enteric adenoviruses in patients from Pune, Aurangabad and Nagpur was found to be 9% (10/111), 7% (7/100), and 7.5% (17/228), respectively. Sequence based analysis of the partial hexon and/or fiber genes showed the presence of adenovirus serotypes 40, 41, and 31 and variations at the subgenus and strain level. Phylogenetic analysis of the adenovirus strains indicated 98-100% homology with adenovirus 40 of the UK, 96-99% with adenovirus 41 of the USA and 94-100% with adenovirus 31 of Austria. The study indicates circulation of enteric adenovirus serotypes 40 and 41 with an unreported serotype 31 in sporadic cases of gastroenteritis. This is the first report from India on the association of enteric adenoviruses with acute gastroenteritis.