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This study was conducted to determine the prevalence of Neospora caninum in dairy & Cross breed of beef cattle and to detect the prevalence of Neospora Oocysts in feces of dogs for the first time in Al-Muthana province-Iraq,800 serum samples of cows were examined by using Indirect Enzyme Linked Immuno Sorbent Assay ( ELISA) kit and 180 dogs fecal samples by direct , flotation test and by PCR assay. The age was ranged between 1-16 years for cows and 3mon- 10years for dogs during the period from March -2010 to June - 2011. The present study recorded the prevalence percent of Hammondia- Neospora like Oocysts (HNLO)that detected in 12 fecal samples 6.66%, then these samples were examined by species specific-PCR, the 3 fecal samples were positive for N. caninum produced visible bands in the PCR product at 337 bp.this work the first time in Al-muthanna province-Iraq about Neosporosis in dogs and cattle, that diagnosed by PCR.Human could become exposed to N.caninum by accidental ingestion of oocyst shed in the feces of canids definitive host or by consumption of undercooked meat that contains tissue cysts and concern is that N.caninum might be zoonotic.