This study conducts to identification the microscopic prevalence of schistosome infection in cattle urine and molecular detection of Schistosoma bovis in urine and blood. Totally, 70 anorexic, emaciated and intermittent diarrheic cattle of different ages and sexes were selected and subjected to collection of urine that sediment firstly and tested by light microscope and the samples blood that examined molecularly using the conventional polymerase chain reaction (PCR) assay. Our findings revealed that 11.43% of study animals were positives by microscopy. Targeting the 18S rRNA gene, the positive findings using the PCR assay detected that (21.43%) of in blood samples were positive; whereas, no positivity was seen in urine. Regarding age, results of microscopy were significantly elevated (P<0.0216) in cattle of >1 year (14.89%) when compared to those of <1 year (4.35%); while molecularly, insignificant variation (P<0.0939) was seen between values of <1 year (21.74%) and >1 year (21.28%). Concerning sex, the findings of microscopy and PCR reported that females (15.39% and 28.21%, respectively) having a higher positivity (P<0.05) than males (6.45% and 129%, respectively). In conclusion, this represents the first Iraqi study indicates traditionally the prevalence of bovine schistosomiasis in urine; and molecularly, S. bovis in blood. The species-specific DNA detection by PCR appears to be more sensitive than clinical and traditional techniques, in addition to its ability in providing a high valuable data about the prevalence of disease. Moreover studies are recommended be done in other Iraqi areas among other domestic animals to explain their role in transmission of infection.