Detection of sperm in the peritoneal fluid of animals is unusual and has not been reported in the literature. In this report, we describe the presence of sperm cells in the peritoneal fluid of a two-year-old ram. The ram was presented with dyspnea, reduced rumen contractions, a mild degree of dehydration, cyanotic mucosa, difficulty in standing, and anuria. Ancillary diagnostics, including ultrasonography, radiography, complete blood cell count, and abdominocentesis were performed. In the peripheral blood sample, no blood parasites were observed, although thrombocytosis with toxic neutrophilia, and atypical lymphocytes were seen in the blood smear. In radiographs, urolithiasis was confirmed just on the sigmoid flexure position. Ultrasound examination revealed a distended bladder and large amounts of free fluid within the peritoneal cavity. The electrocardiogram analysis showed the absence of P waves, bradycardia, wide QRS complexes, ST-segment elevation, and tented T waves. In the peritoneal fluid smears, a large number of spermatozoa, and increased inflammatory cells were observed. It was concluded that the trauma or rupture in colliculus seminalis, ductus deferens, or the urethra with urinary stones resulted in leakage of spermatozoa to the peritoneal cavity. To our knowledge, this is the first report of the presence of spermatozoa in the peritoneal fluid of large animals.