First molecular characterization and morphological aspects of Sarcocystis fusiformis infecting water buffalo Bubalus bubalis in Egypt

Fresh muscle samples from water buffalo (Bubalus bubalis) aged 2-15, from Giza Province, Egypt; were examined for Sarcocystis infection. Macroscopic ovoid sarcocysts embedded in the muscle tissues of the examined buffaloes were detected; they measured 152-230 (210 +/- 7) mu m in length and 37-119 (95 +/- 3) mu m in width. The esophagus was the most infected organ followed by the diaphragm, and tongue, while the heart muscles were the least infected. The cyst cavity was compartmentalized by septa derived from the ground substance located under the primary cyst wall. Using transmission electron microscopy, the primary cyst wall bordered sarcocysts were determined to be 0.08-0.22 mu m in thickness, raised from the parasitophorous vacuolar membrane, and surrounded by a secondary cyst wall of host origin. The primary cyst wall had irregular wall folds with numerous cauliflower-like projections of variable sizes and shapes accompanied by knob-like electron-dense elevations. 18S rRNA gene expression studies confirmed that the present parasite isolates belonged to the genus Sarcocystis. The sequence data showed significant identities (>90%) with archived gene sequences from many Eimeriidae organisms, and a dendogram showing the phylogenetic relationship was constructed. The most closely related species was Sarcocystis fusiformis KR186117, with an identity percentage of 98%. The recovered sequences were deposited in the GenBank under the accession number MG572125. The present study, to our knowledge, is the first collective ultrastructural and molecular study that confirmed the taxonomy of sarcocysts isolated from water buffaloes in Egypt as Sarcocystis fusiformis.