Abstract: Objective To establish an infection model and nested PCR assay method for Eimeria stiedai (E.stiedai) in rabbit. Methods The established infection model of E.stiedai in rabbits was identified by clinical examination, microscopic observation, blood biochemistry and histopathology. By collecting oocysts, DNA of E.stiedai was extracted, and the specific primers were designed to establish nested PCR for detection of E.stiedai. Results Clinical necropsy: the abdominal circumference of the rabbit was obviously enlarged. The liver enlargement and a large number of white and light yellow nodules were found by biopsy, while the gallbladder and bile duct were swollen, and the bile was pale yellow. Microscopic examination: the size of the eggs was (31.72-38.43) ?m×(18.10-22.69) ?m. Blood biochemical examination: the content of globulin (GLOB) was increased, while the levels of creatinine (CREA) and alkaline phosphatase (ALKP) were decreased, and the rest of the detection indicators were in the reference range. Histopathological examination: a large number of pink E.stiedai eggs were seen in the liver tissue and the bile duct. Nested PCR: the lower limit of detection was 1 oocyst DNA sample and 1.14×10³ copies of plasmids, and the negative control and blank control showed no bands. The variability coefficient was less than 5%. Conclusion The E.stiedai infection model in rabbits can be successfully constructed, and the nested PCR assay can amplify the specific fragments of E.stiedai with high sensitivity and good repeatability.

Key words: Eimeria stiedai, Histopathology, Nested PCR, Rabbits