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Faecal samples from 183 dogs living in three different shelters in the Rome metropolitan area were randomly collected and examined for the prevalence of giardiosis. Giardia infections were detected by a commercially available ELISA test (ProspecT Giardia Microplate Assay). Overall prevalence was 55.2%. Prevalence rates in single shelters were 74.3, 35.5, and 20.9%, respectively. Using multivariate analysis, no association was found between Giardia-positivity and shelter or sex, breed, or diarrhoea. Giardia-positive dogs were more likely to be younger than 5-year-old (odds ratio [OR] = 2.87; 95% confidence interval [95% CI]: 1.07-7.77; p = 0.038), living together (OR = 2.58; 95% CI: 1.12-5.93; p = 0.026), and fed commercial wet food, both alone and combined with dry food (OR = 5.67; 95% CI: 1.59-20.24; p = 0.008). Correlation between Giardia infection and type of food has not been previously reported in dogs. Possible use of the ELISA test for detection of Giardia infection in dogs and zoonotic implications are discussed.