The molecular prevalence and genotypes ofGiardia duodenalisin cattle were investigated. A total of 450 fecal samples were collected from cattle in three provinces of Central Anatolia from August 2017 to July 2019. Genomic DNA was extracted from the fecal samples and used in molecular analysis carried out by nested PCR analyses of the beta-giardin(bg) gene ofG. duodenalis. Positive samples were further analyzed by nested PCR at two gene loci (triosephosphate isomerase(tpi) andglutamate dehydrogenase(gdh)) for genotyping ofG. duodenalisisolates. PCR analyses of thebggene indicated that the overall prevalence ofG. duodenaliswas 30.2%. However, lower rates were determined with PCR analyses forgdhandtpiloci. The sequence analyses of thebg,gdh, andtpigenes revealed the presence of zoonotic assemblage A and livestock-specific assemblage E. Combined-sequence analyses revealed that assemblage E was the most common in the study area. Our study provides the first data on the wide prevalence of livestock-specific assemblages E in cattle in Turkey. The prevalence of assemblage A in cattle also reveals the importance of cattle for zoonotic transmission of giardiasis in Turkey.